49 results on '"Seeger, Werner"'
Search Results
2. Evidence for the Fucoidan/P-Selectin Axis as a Therapeutic Target in Hypoxia-induced Pulmonary Hypertension.
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Novoyatleva T, Kojonazarov B, Owczarek A, Veeroju S, Rai N, Henneke I, Böhm M, Grimminger F, Ghofrani HA, Seeger W, Weissmann N, and Schermuly RT
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- Animals, Biomarkers blood, Disease Models, Animal, Fucus chemistry, Humans, Hypertension, Pulmonary physiopathology, Hypoxia physiopathology, Male, Mice, Anticoagulants therapeutic use, Hypertension, Pulmonary drug therapy, Hypertension, Pulmonary etiology, Hypoxia complications, Myocytes, Smooth Muscle drug effects, P-Selectin blood, Polysaccharides therapeutic use
- Abstract
Rationale: Pulmonary arterial hypertension (PAH) is characterized by vascular remodeling and excessive proliferation of pulmonary artery smooth muscle cells (PASMCs). Fucoidan, a polysaccharidic ligand of the adhesion molecule P-selectin, exhibits antiproliferative properties. The effects of the fucoidan/P-selectin axis on vascular remodeling and pulmonary hypertension (PH) after hypoxia remain unexplored. Objectives: We aimed to evaluate the therapeutic potential of targeting the fucoidan/P-selectin axis in PH. Methods: Mice with PH induced by chronic hypoxia (35 d) were given either fucoidan (from Fucus vesiculosus ) or anti-P-selectin antibody (Rb40.34) during Days 21-35. Right ventricular (RV) function was determined by echocardiography. Vascular morphometry was assessed by immunohistochemistry. Human and experimental PH lungs and PASMCs were used for assessment of P-selectin expression and function. Measurements and Main Results: Fucoidan attenuated chronic hypoxia-induced PH in mice, reducing pulmonary vascular remodeling and restoring RV function. In vitro , fucoidan inhibited hypoxia and growth factor-stimulated PASMC proliferation and migration. Chronic hypoxia caused an upregulation of P-selectin in the medial layer of the small pulmonary arteries. P-selectin was persistently upregulated in PASMCs of human and hypoxia-induced experimental PH. HIF-1α (hypoxia-inducible factor 1α) directly bound to the P-selectin promoter and transcriptionally activated P-selectin in hypoxia. P-selectin blockage resulted in a marked reduction of PASMC proliferation in vitro . Blockage of P-selectin by administration of anti-P-selectin Rb40.34 antibody and P-selectin-deficient mice improved vascular remodeling and restored RV function. Conclusions: Fucoidan is a potent natural adjuvant that represents a promising therapeutic approach for PH. Our data indicate a previously unrecognized role of P-selectin in the proliferative response of PASMCs associated with PH.
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- 2019
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3. Pulmonary hypertension in chronic lung disease and hypoxia.
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Nathan SD, Barbera JA, Gaine SP, Harari S, Martinez FJ, Olschewski H, Olsson KM, Peacock AJ, Pepke-Zaba J, Provencher S, Weissmann N, and Seeger W
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- Animals, Antihypertensive Agents therapeutic use, Chronic Disease, Humans, Hypertension, Pulmonary drug therapy, Lung Diseases, Interstitial drug therapy, Lung Diseases, Interstitial pathology, Pulmonary Disease, Chronic Obstructive drug therapy, Hypertension, Pulmonary epidemiology, Hypoxia complications, Lung Diseases, Interstitial complications, Pulmonary Disease, Chronic Obstructive complications
- Abstract
Pulmonary hypertension (PH) frequently complicates the course of patients with various forms of chronic lung disease (CLD). CLD-associated PH (CLD-PH) is invariably associated with reduced functional ability, impaired quality of life, greater oxygen requirements and an increased risk of mortality. The aetiology of CLD-PH is complex and multifactorial, with differences in the pathogenic sequelae between the diverse forms of CLD. Haemodynamic evaluation of PH severity should be contextualised within the extent of the underlying lung disease, which is best gauged through a combination of physiological and imaging assessment. Who, when, if and how to screen for PH will be addressed in this article, as will the current state of knowledge with regard to the role of treatment with pulmonary vasoactive agents. Although such therapy cannot be endorsed given the current state of findings, future studies in this area are strongly encouraged., Competing Interests: Conflict of interest: S.D. Nathan is a consultant for and has received research funding from Bellerophon, United Therapeutics and Bayer Pharmaceuticals; is a consultant for Third Pole and Actelion, and is a consultant for, has received research funding from and is on the speakers’ bureau of Roche-Genentech and Boehringer Ingelheim. Conflict of interest: J.A. Barbera reports grants and personal fees from Actelion and MSD, personal fees from Arena, and grants from Bayer and GSK, outside the submitted work. Conflict of interest: S.P. Gaine reports personal fees from Actelion, United Therapeutics, MSD and GSK, outside the submitted work. Conflict of interest: S. Harari has received grants for research and speakers fees from Actelion, Boehringer Ingelheim and Roche. Conflict of interest: F.J. Martinez has received grants from the NIH (IPF UO1, COPD UO1/RO1); personal fees, honoraria and non-personal travel support from the American College of Chest Physicians, Continuing Education, ConCert, Inova Fairfax Health System, MD Magazine, Miller Communications, National Association for Continuing Education, Novartis, Pearl Pharmaceuticals, PeerView Communications, Prime Communications, Puerto Rican Respiratory Society, Roche, Sunovion, Theravance, Potomac, University of Alabama Birmingham and Zambon; personal fees and non-personal or non-financial travel support from AstraZeneca; personal fees, non-personal travel support, and non-financial support for data and safety monitoring board work from Boehringer Ingelheim; personal fees, honoraria, non-personal travel support, and non-financial support for data and safety monitoring board work from Genentech and GlaxoSmithKline; personal fees and honoraria from Columbia University, Integritas, Methodist Hospital Brooklyn, New York University, Unity, UpToDate, WebMD/MedScape, Western Connecticut Health Network, Academic CME, Patara, PlatformIQ, American Thoracic Society, Rockpointe and Rare Disease Healthcare Communications; non-personal travel support from Nitto; personal fees, honoraria, travel support and non-personal travel support from Chiesi; personal fees, honoraria and travel support from Physicians Education Resource and Teva; honoraria and travel support from Canadian Respiratory Network; personal fees from France Foundation; and has participated on scientific advisory boards (no direct financial compensation) for ProterrixBio and Bridge Biotherapeutics; participated on IPF study steering committees (no direct financial compensation) for Afferent/Merck, Gilead, Veracyte, Prometic, Bayer and ProMedior; and participated on an IPF study steering committee and data safety monitoring board (no direct financial compensation) for Biogen. Conflict of interest: H. Olschewski reports personal fees and non-financial support from Bayer, MSD, Pfizer and Novartis, grants, personal fees and non-financial support from Actelion, grants from Inventiva, and personal fees from Bellerophon, outside the submitted work; and is part-time employee of the Ludwig Boltzmann Institute for Lung Vascular Research. Conflict of interest: K.M. Olsson received fees for talks and consulting work from Actelion, Bayer, GSK, Pfizer and United Therapeutics. Conflict of interest: A.J. Peacock has received research grants and personal fees from Actelion Pharmaceuticals, Bayer, GSK, MSD, Pfizer and United Therapeutics, outside the submitted work. Conflict of interest: J. Pepke-Zaba is a member of the advisory boards for Actelion, Merck, Bayer and GSK, has received grants, personal fees and non-financial support from Actelion, Merck and Bayer, and personal fees from GSK. Conflict of interest: S. Provencher has received research grants from Actelion Pharmaceuticals and Boehringer Ingelheim, and has received speaker fees from Actelion Pharmaceuticals. Conflict of interest: N. Weissmann has nothing to disclose. Conflict of interest: W. Seeger has received consultancy fees from Bayer AG, United Therapeutics, Liquidia, Vectura and Novartis., (Copyright ©ERS 2019.)
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- 2019
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4. miR-223-IGF-IR signalling in hypoxia- and load-induced right-ventricular failure: a novel therapeutic approach.
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Shi L, Kojonazarov B, Elgheznawy A, Popp R, Dahal BK, Böhm M, Pullamsetti SS, Ghofrani HA, Gödecke A, Jungmann A, Katus HA, Müller OJ, Schermuly RT, Fisslthaler B, Seeger W, and Fleming I
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- Animals, Gene Expression Regulation, Genetic Predisposition to Disease, Heart Failure genetics, Heart Failure physiopathology, Heart Failure prevention & control, Heart Ventricles physiopathology, Humans, Hypertension, Pulmonary genetics, Hypertension, Pulmonary physiopathology, Hypertension, Pulmonary prevention & control, Hypertrophy, Right Ventricular metabolism, Hypertrophy, Right Ventricular physiopathology, Hypoxia complications, Hypoxia genetics, Hypoxia-Inducible Factor 1, alpha Subunit genetics, Hypoxia-Inducible Factor 1, alpha Subunit metabolism, Imidazoles pharmacology, Male, Mice, Inbred C57BL, Mice, Knockout, MicroRNAs genetics, Phenotype, Pyridines pharmacology, Receptor, IGF Type 1 antagonists & inhibitors, Receptor, IGF Type 1 deficiency, Receptor, IGF Type 1 genetics, Signal Transduction, Ventricular Dysfunction, Right genetics, Ventricular Dysfunction, Right physiopathology, Ventricular Dysfunction, Right prevention & control, Heart Failure metabolism, Heart Ventricles metabolism, Hypertension, Pulmonary metabolism, Hypoxia metabolism, Lung metabolism, MicroRNAs metabolism, Receptor, IGF Type 1 metabolism, Receptors, Somatomedin metabolism, Ventricular Dysfunction, Right metabolism, Ventricular Function, Right
- Abstract
Aims: Pulmonary hypertension is a progressive disease with poor prognosis, characterized by pathological inward remodelling and loss of patency of the lung vasculature. The right ventricle is co-affected by pulmonary hypertension, which triggers events such as hypoxia and/or increased mechanical load. Initially the right ventricle responds with 'adaptive' hypertrophy, which is often rapidly followed by 'maladaptive' changes leading to right heart decompensation and failure, which is the ultimate cause of death., Methods and Results: We report here that miR-223 is expressed in the murine lung and right ventricle at higher levels than in the left ventricle. Moreover, lung and right-ventricular miR-223 levels were markedly down-regulated by hypoxia. Correspondingly, increasing right-ventricular load by pulmonary artery banding, induced right-ventricular ischaemia, and the down-regulation of miR-223. Lung and right ventricle miR-223 down-regulation were linked with increased expression of the miR-223 target; insulin-like growth factor-I receptor (IGF-IR) and IGF-I downstream signalling. Similarly, miR-223 was decreased and IGF-IR increased in human pulmonary hypertension. Notably in young mice, miR-223 overexpression, the genetic inactivation or pharmacological inhibition of IGF-IR, all attenuated right-ventricular hypertrophy and improved right heart function under conditions of hypoxia or increased afterload., Conclusion: These findings highlight the early role of pulmonary and right-ventricular miR-223 and the IGF-IR in the right heart failure programme initiated by pulmonary hypoxia and increased mechanical load and may lead to the development of novel therapeutic strategies that target the development of PH and right heart failure., (Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2016. For permissions please email: journals.permissions@oup.com.)
- Published
- 2016
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5. Structural and functional prevention of hypoxia-induced pulmonary hypertension by individualized exercise training in mice.
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Weissmann N, Peters DM, Klöpping C, Krüger K, Pilat C, Katta S, Seimetz M, Ghofrani HA, Schermuly RT, Witzenrath M, Seeger W, Grimminger F, and Mooren FC
- Subjects
- 3',5'-Cyclic-AMP Phosphodiesterases genetics, 3',5'-Cyclic-AMP Phosphodiesterases metabolism, Animals, Exercise Therapy, Exercise Tolerance, Gene Expression, Hypertension, Pulmonary etiology, Hypoxia complications, Lung blood supply, Lung metabolism, Male, Mice, Mice, Inbred C57BL, Muscle, Smooth, Vascular physiopathology, Nitric Oxide Synthase genetics, Nitric Oxide Synthase metabolism, Oxygen Consumption, Phosphodiesterase 5 Inhibitors pharmacology, Physical Conditioning, Animal, Piperazines pharmacology, Purines pharmacology, Signal Transduction, Sildenafil Citrate, Sulfones pharmacology, Ventricular Pressure, Hypertension, Pulmonary prevention & control, Hypoxia therapy
- Abstract
Pulmonary hypertension (PH) is a disease with a poor prognosis characterized by a vascular remodeling process and an increase in pulmonary vascular resistance. While a variety of reports demonstrated that exercise training exerts beneficial effects on exercise performance and quality of life in PH patients, it is not known how physical exercise affects vascular remodeling processes occurring in hypoxia-induced PH. Therefore, we investigated the effect of individualized exercise training on the development of hypoxia-induced PH in mice. Training effects were compared with pharmacological treatment with the phosphodiesterase 5 inhibitor Sildenafil or a combination of training plus Sildenafil. Trained mice who received Sildenafil showed a significantly improved walking distance (from 88.9 ± 8.1 to 146.4 ± 13.1 m) and maximum oxygen consumption (from 93.3 ± 2.9 to 105.5 ± 2.2% in combination with Sildenafil, to 102.2 ± 3.0% with placebo) compared with sedentary controls. Right ventricular systolic pressure, measured by telemetry, was at the level of healthy normoxic animals, whereas right heart hypertrophy did not benefit from training. Most interestingly, the increase in small pulmonary vessel muscularization was prevented by training. Respective counterregulatory processes were detected for the nitric oxide-soluble guanylate cyclase-phosphodiesterase system. We conclude that individualized daily exercise can prevent vascular remodeling in hypoxia-induced PH., (Copyright © 2014 the American Physiological Society.)
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- 2014
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6. Classical transient receptor potential channel 1 in hypoxia-induced pulmonary hypertension.
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Malczyk M, Veith C, Fuchs B, Hofmann K, Storch U, Schermuly RT, Witzenrath M, Ahlbrecht K, Fecher-Trost C, Flockerzi V, Ghofrani HA, Grimminger F, Seeger W, Gudermann T, Dietrich A, and Weissmann N
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- Animals, Biomarkers metabolism, Blotting, Western, Cells, Cultured, Chronic Disease, Female, Hypertension, Pulmonary etiology, Hypertension, Pulmonary pathology, Lung metabolism, Lung pathology, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Pulmonary Artery metabolism, Pulmonary Artery pathology, Real-Time Polymerase Chain Reaction, TRPC Cation Channels deficiency, Up-Regulation, Hypertension, Pulmonary metabolism, Hypoxia complications, TRPC Cation Channels metabolism
- Abstract
Rationale: Pulmonary hypertension (PH) is a life-threatening disease, characterized by pulmonary vascular remodeling. Abnormal smooth muscle cell proliferation is a primary hallmark of chronic hypoxia-induced PH. Essential for cell growth are alterations in the intracellular Ca(2+) homeostasis. Classical transient receptor potential (TRPC) proteins have been suggested to contribute to PH development, as TRPC1 and TRPC6 are predominantly expressed in precapillary pulmonary arterial smooth muscle cells (PASMC). Studies in a TRPC6-deficient mouse model revealed an essential function of TRPC6 in acute but not in chronic hypoxia., Objectives: We aimed to identify the importance of TRPC1 in the pathogenesis of chronic hypoxia-induced PH in mice., Methods: TRPC1 expression analysis was performed using real-time polymerase chain reaction. TRPC1 function was assessed by in vivo experiments in TRPC1(-/-) animals as well as in isolated precapillary murine PASMC after TRPC1 knockdown by TRPC1-specific small interfering RNAs., Measurements and Main Results: Only TRPC1 mRNA was up-regulated under hypoxia in isolated murine PASMC (1% O2 for 72 h). Hypoxia-induced proliferation of murine PASMC was attenuated in cells treated with small interfering RNA against TRPC1 and in cells isolated from TRPC1(-/-) animals compared with untreated and wild-type cells. TRPC1(-/-) mice did not develop PH in response to chronic hypoxia (FI(O2) 0.10 for 21 d) and had less vascular muscularization but a similar degree of right ventricular hypertrophy compared with wild-type mice., Conclusions: Our results indicate an important role of TRPC1 in pulmonary vascular remodeling underlying the development of hypoxia-induced PH.
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- 2013
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7. Effects of dimethylarginine dimethylaminohydrolase-1 overexpression on the response of the pulmonary vasculature to hypoxia.
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Bakr A, Pak O, Taye A, Hamada F, Hemeida R, Janssen W, Gierhardt M, Ghofrani HA, Seeger W, Grimminger F, Schermuly RT, Witzenrath M, Brandes RP, Huang N, Cooke JP, Weissmann N, and Sommer N
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- Amidohydrolases metabolism, Animals, Arginine analogs & derivatives, Blood Vessels drug effects, Blood Vessels physiopathology, Cyclic GMP metabolism, Gene Expression, Hemodynamics drug effects, Hypertension, Pulmonary etiology, Hypertension, Pulmonary metabolism, Hypertension, Pulmonary physiopathology, Hypoxia complications, Hypoxia metabolism, Hypoxia physiopathology, Lung drug effects, Lung physiopathology, Mice, Nitroarginine pharmacology, Organ Culture Techniques, Oxadiazoles pharmacology, Signal Transduction, Vasoconstriction drug effects, Amidohydrolases genetics, Arginine metabolism, Hypertension, Pulmonary genetics, Hypoxia genetics, Lung metabolism, Nitric Oxide metabolism
- Abstract
Acute and sustained hypoxic pulmonary vasoconstriction (HPV), as well as chronic pulmonary hypertension (PH), is modulated by nitric oxide (NO). NO synthesis can be decreased by asymmetric dimethylarginine (ADMA), which is degraded by dimethylarginine dimethylaminohydrolase-1 (DDAH1). We investigated the effects of DDAH1 overexpression (DDAH1(tg)) on HPV and chronic hypoxia-induced PH. HPV was measured during acute (10 min) and sustained (3 h) hypoxia in isolated mouse lungs. Chronic PH was induced by the exposure of mice to 4 weeks of hypoxia. ADMA and cyclic 3',5'-guanosine monophosphate (cGMP) were determined by ELISA, and NO generation was determined by chemiluminescence. DDAH1 overexpression exerted no effects on acute HPV. However, DDAH1(tg) mice showed decreased sustained HPV compared with wild-type (WT) mice. Concomitantly, ADMA was decreased, and concentrations of NO and cGMP were significantly increased in DDAH1(tg). The administration of either Nω-nitro-l-arginine or 1H-[1,2,4]oxadiazolo [4,3-a]quinoxalin-1-one potentiated sustained HPV and partly abolished the differences in sustained HPV between WT and DDAH1(tg) mice. The overexpression of DDAH1 exerted no effect on the development of chronic hypoxia-induced PH. DDAH1 overexpression selectively decreased the sustained phase of HPV, partly via activation of the NO-cGMP pathway. Thus, increased ADMA concentrations modulate sustained HPV, but not acute HPV or chronic hypoxia-induced PH.
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- 2013
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8. Hypoxia induces Kv channel current inhibition by increased NADPH oxidase-derived reactive oxygen species.
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Mittal M, Gu XQ, Pak O, Pamenter ME, Haag D, Fuchs DB, Schermuly RT, Ghofrani HA, Brandes RP, Seeger W, Grimminger F, Haddad GG, and Weissmann N
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- Acetophenones pharmacology, Animals, Cells, Cultured, Hypertension, Pulmonary etiology, Hypoxia complications, Male, Mice, Myocytes, Smooth Muscle drug effects, Myocytes, Smooth Muscle pathology, NADPH Oxidase 4, NADPH Oxidases genetics, Oxidation-Reduction drug effects, Protein Transport, Pulmonary Artery pathology, RNA, Small Interfering genetics, Rats, Rats, Sprague-Dawley, Reactive Oxygen Species metabolism, Hypertension, Pulmonary metabolism, Hypoxia metabolism, Kv1.5 Potassium Channel metabolism, Myocytes, Smooth Muscle metabolism, NADPH Oxidases metabolism
- Abstract
There is current discussion whether reactive oxygen species are up- or downregulated in the pulmonary circulation during hypoxia, from which sources (i.e., mitochondria or NADPH oxidases) they are derived, and what the downstream targets of ROS are. We recently showed that the NADPH oxidase homolog NOX4 is upregulated in hypoxia-induced pulmonary hypertension in mice and contributes to the vascular remodeling in pulmonary hypertension. We here tested the hypothesis that NOX4 regulates K(v) channels via an increased ROS formation after prolonged hypoxia. We showed that (1) NOX4 is upregulated in hypoxia-induced pulmonary hypertension in rats and isolated rat pulmonary arterial smooth muscle cells (PASMC) after 3days of hypoxia, and (2) that NOX4 is a major contributor to increased reactive oxygen species (ROS) after hypoxia. Our data indicate colocalization of K(v)1.5 and NOX4 in isolated PASMC. The NADPH oxidase inhibitor and ROS scavenger apocynin as well as NOX4 siRNA reversed the hypoxia-induced decrease in K(v) current density whereas the protein levels of the channels remain unaffected by siNOX4 treatment. Determination of cysteine oxidation revealed increased NOX4-mediated K(v)1.5 channel oxidation. We conclude that sustained hypoxia decreases K(v) channel currents by a direct effect of a NOX4-derived increase in ROS., (Copyright © 2011 Elsevier Inc. All rights reserved.)
- Published
- 2012
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9. Effects of hypercapnia and NO synthase inhibition in sustained hypoxic pulmonary vasoconstriction.
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Ketabchi F, Ghofrani HA, Schermuly RT, Seeger W, Grimminger F, Egemnazarov B, Shid-Moosavi SM, Dehghani GA, Weissmann N, and Sommer N
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- Acidosis drug therapy, Acidosis physiopathology, Animals, Enzyme Inhibitors pharmacology, Hypercapnia drug therapy, Hypoxia drug therapy, Imines pharmacology, Lung blood supply, Lung drug effects, Male, Nitric Oxide Synthase Type II antagonists & inhibitors, Nitric Oxide Synthase Type III antagonists & inhibitors, Nitroarginine pharmacology, Pulmonary Circulation drug effects, Pulmonary Circulation physiology, Rabbits, Sodium Bicarbonate pharmacology, Vasoconstriction drug effects, Hypercapnia physiopathology, Hypoxia physiopathology, Lung physiopathology, Nitric Oxide Synthase Type II physiology, Nitric Oxide Synthase Type III physiology, Vasoconstriction physiology
- Abstract
Background: Acute respiratory disorders may lead to sustained alveolar hypoxia with hypercapnia resulting in impaired pulmonary gas exchange. Hypoxic pulmonary vasoconstriction (HPV) optimizes gas exchange during local acute (0-30 min), as well as sustained (> 30 min) hypoxia by matching blood perfusion to alveolar ventilation. Hypercapnia with acidosis improves pulmonary gas exchange in repetitive conditions of acute hypoxia by potentiating HPV and preventing pulmonary endothelial dysfunction. This study investigated, if the beneficial effects of hypercapnia with acidosis are preserved during sustained hypoxia as it occurs, e.g in permissive hypercapnic ventilation in intensive care units. Furthermore, the effects of NO synthase inhibitors under such conditions were examined., Method: We employed isolated perfused and ventilated rabbit lungs to determine the influence of hypercapnia with or without acidosis (pH corrected with sodium bicarbonate), and inhibitors of endothelial as well as inducible NO synthase on acute or sustained HPV (180 min) and endothelial permeability., Results: In hypercapnic acidosis, HPV was intensified in sustained hypoxia, in contrast to hypercapnia without acidosis when HPV was amplified during both phases. L-NG-Nitroarginine (L-NNA), a non-selective NO synthase inhibitor, enhanced acute as well as sustained HPV under all conditions, however, the amplification of sustained HPV induced by hypercapnia with or without acidosis compared to normocapnia disappeared. In contrast 1400 W, a selective inhibitor of inducible NO synthase (iNOS), decreased HPV in normocapnia and hypercapnia without acidosis at late time points of sustained HPV and selectively reversed the amplification of sustained HPV during hypercapnia without acidosis. Hypoxic hypercapnia without acidosis increased capillary filtration coefficient (Kfc). This increase disappeared after administration of 1400 W., Conclusion: Hypercapnia with and without acidosis increased HPV during conditions of sustained hypoxia. The increase of sustained HPV and endothelial permeability in hypoxic hypercapnia without acidosis was iNOS dependent.
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- 2012
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10. Diacylglycerol regulates acute hypoxic pulmonary vasoconstriction via TRPC6.
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Fuchs B, Rupp M, Ghofrani HA, Schermuly RT, Seeger W, Grimminger F, Gudermann T, Dietrich A, and Weissmann N
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- 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid pharmacology, Animals, Diacylglycerol Kinase antagonists & inhibitors, Diacylglycerol Kinase metabolism, Dose-Response Relationship, Drug, Enzyme Inhibitors pharmacology, Estrenes pharmacology, Hypoxia genetics, Hypoxia physiopathology, In Vitro Techniques, Mice, Mice, Inbred C57BL, Mice, Knockout, Perfusion, Piperidines pharmacology, Pyrrolidinones pharmacology, Quinazolinones pharmacology, Signal Transduction, TRPC Cation Channels deficiency, TRPC Cation Channels genetics, TRPC6 Cation Channel, Time Factors, Type C Phospholipases antagonists & inhibitors, Type C Phospholipases metabolism, Vasoconstrictor Agents pharmacology, Diglycerides metabolism, Hypoxia metabolism, Lung blood supply, Pulmonary Circulation drug effects, TRPC Cation Channels metabolism, Vasoconstriction drug effects
- Abstract
Background: Hypoxic pulmonary vasoconstriction (HPV) is an essential mechanism of the lung that matches blood perfusion to alveolar ventilation to optimize gas exchange. Recently we have demonstrated that acute but not sustained HPV is critically dependent on the classical transient receptor potential 6 (TRPC6) channel. However, the mechanism of TRPC6 activation during acute HPV remains elusive. We hypothesize that a diacylglycerol (DAG)-dependent activation of TRPC6 regulates acute HPV., Methods: We investigated the effect of the DAG analog 1-oleoyl-2-acetyl-sn-glycerol (OAG) on normoxic vascular tone in isolated perfused and ventilated mouse lungs from TRPC6-deficient and wild-type mice. Moreover, the effects of OAG, the DAG kinase inhibitor R59949 and the phospholipase C inhibitor U73122 on the strength of HPV were investigated compared to those on non-hypoxia-induced vasoconstriction elicited by the thromboxane mimeticum U46619., Results: OAG increased normoxic vascular tone in lungs from wild-type mice, but not in lungs from TRPC6-deficient mice. Under conditions of repetitive hypoxic ventilation, OAG as well as R59949 dose-dependently attenuated the strength of acute HPV whereas U46619-induced vasoconstrictions were not reduced. Like OAG, R59949 mimicked HPV, since it induced a dose-dependent vasoconstriction during normoxic ventilation. In contrast, U73122, a blocker of DAG synthesis, inhibited acute HPV whereas U73343, the inactive form of U73122, had no effect on HPV., Conclusion: These findings support the conclusion that the TRPC6-dependency of acute HPV is induced via DAG.
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- 2011
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11. Nebulization of the acidified sodium nitrite formulation attenuates acute hypoxic pulmonary vasoconstriction.
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Egemnazarov B, Schermuly RT, Dahal BK, Elliott GT, Hoglen NC, Surber MW, Weissmann N, Grimminger F, Seeger W, and Ghofrani HA
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- Acute Disease, Administration, Inhalation, Animals, Blood Pressure drug effects, Cardiac Output drug effects, Chemistry, Pharmaceutical, Disease Models, Animal, Dose-Response Relationship, Drug, Exhalation, Hydrogen-Ion Concentration, Hypoxia physiopathology, Male, Nebulizers and Vaporizers, Nitrates blood, Nitric Oxide metabolism, Perfusion, Pulmonary Artery physiopathology, Rabbits, Time Factors, Hypoxia drug therapy, Pulmonary Artery drug effects, Sodium Nitrite administration & dosage, Vasoconstriction drug effects, Vasodilator Agents administration & dosage
- Abstract
Background: Generalized hypoxic pulmonary vasoconstriction (HPV) occurring during exposure to hypoxia is a detrimental process resulting in an increase in lung vascular resistance. Nebulization of sodium nitrite has been shown to inhibit HPV. The aim of this project was to investigate and compare the effects of nebulization of nitrite and different formulations of acidified sodium nitrite on acute HPV., Methods: Ex vivo isolated rabbit lungs perfused with erythrocytes in Krebs-Henseleit buffer (adjusted to 10% hematocrit) and in vivo anesthetized catheterized rabbits were challenged with periods of hypoxic ventilation alternating with periods of normoxic ventilation. After baseline hypoxic challenges, vehicle, sodium nitrite or acidified sodium nitrite was delivered via nebulization. In the ex vivo model, pulmonary arterial pressure and nitric oxide concentrations in exhaled gas were monitored. Nitrite and nitrite/nitrate were measured in samples of perfusion buffer. Pulmonary arterial pressure, systemic arterial pressure, cardiac output and blood gases were monitored in the in vivo model., Results: In the ex vivo model, nitrite nebulization attenuated HPV and increased nitric oxide concentrations in exhaled gas and nitrite concentrations in the perfusate. The acidified forms of sodium nitrite induced higher levels of nitric oxide in exhaled gas and had longer vasodilating effects compared to nitrite alone. All nitrite formulations increased concentrations of circulating nitrite to the same degree. In the in vivo model, inhaled nitrite inhibited HPV, while pulmonary arterial pressure, cardiac output and blood gases were not affected. All nitrite formulations had similar potency to inhibit HPV. The tested concentration of appeared tolerable., Conclusion: Nitrite alone and in acidified forms effectively and similarly attenuates HPV. However, acidified nitrite formulations induce a more pronounced increase in nitric oxide exhalation.
- Published
- 2010
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12. Effects of hypercapnia with and without acidosis on hypoxic pulmonary vasoconstriction.
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Ketabchi F, Egemnazarov B, Schermuly RT, Ghofrani HA, Seeger W, Grimminger F, Shid-Moosavi M, Dehghani GA, Weissmann N, and Sommer N
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- Acidosis, Respiratory physiopathology, Animals, Capillary Permeability drug effects, Endothelium, Vascular drug effects, Endothelium, Vascular physiopathology, Hydrogen-Ion Concentration drug effects, In Vitro Techniques, Lung drug effects, Lung physiopathology, Male, Metalloporphyrins pharmacology, Nitric Oxide metabolism, Nitroarginine pharmacology, Partial Pressure, Pulmonary Ventilation drug effects, Rabbits, Vasoconstriction drug effects, Acidosis, Respiratory complications, Hypercapnia complications, Hypercapnia physiopathology, Hypoxia complications, Hypoxia physiopathology, Lung blood supply, Vasoconstriction physiology
- Abstract
Acute respiratory disorders and permissive hypercapnic strategy may lead to alveolar hypoxia and hypercapnic acidosis. However, the effects of hypercapnia with or without acidosis on hypoxic pulmonary vasoconstriction (HPV) and oxygen diffusion capacity of the lung are controversial. We investigated the effects of hypercapnic acidosis and hypercapnia with normal pH (pH corrected with sodium bicarbonate) on HPV, capillary permeability, gas exchange, and ventilation-perfusion matching in the isolated ventilated-perfused rabbit lung. No alteration in vascular tone was noted during normoxic hypercapnia with or without acidosis compared with normoxic normocapnia. Hypercapnia with normal pH resulted in a transient increase in HPV during the course of consecutive ventilation maneuvers, whereas hypercapnic acidosis increased HPV over time. Hypercapnic acidosis decreased exhaled NO during hypoxia more than hypercapnia with normal pH and normocapnia, whereas intravascular NO release was unchanged. However, inhibition of NO synthesis by nitro-L-arginine (L-NNA) resulted in a loss of the increased HPV caused by hypercapnic acidosis but not that caused by hypercapnia with normal pH. Furthermore, capillary permeability increased during hypoxic hypercapnia with normal pH but not hypoxic hypercapnic acidosis. This effect was NO-dependent because it disappeared during L-NNA administration. Ventilation-perfusion matching and arterial PO2 were improved according to the strength of HPV in hypercapnia compared with normocapnia during Tween nebulization-induced lung injury. In conclusion, the increased HPV during hypercapnic acidosis is beneficial to lung gas exchange by improving ventilation-perfusion matching and preserving the capillary barrier function. These effects seem to be linked to NO-mediated pathways.
- Published
- 2009
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13. The soluble guanylate cyclase activator HMR1766 reverses hypoxia-induced experimental pulmonary hypertension in mice.
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Weissmann N, Hackemack S, Dahal BK, Pullamsetti SS, Savai R, Mittal M, Fuchs B, Medebach T, Dumitrascu R, Eickels Mv, Ghofrani HA, Seeger W, Grimminger F, and Schermuly RT
- Subjects
- Animals, Cardiomegaly, Cyclic GMP metabolism, Hemodynamics drug effects, Hypertension, Pulmonary enzymology, Mice, Myocytes, Smooth Muscle drug effects, Myocytes, Smooth Muscle metabolism, Pulmonary Artery cytology, Pulmonary Artery drug effects, Pulmonary Artery metabolism, Soluble Guanylyl Cyclase, Superoxides metabolism, Vasoconstriction drug effects, Guanylate Cyclase metabolism, Hypertension, Pulmonary prevention & control, Hypoxia prevention & control, Receptors, Cytoplasmic and Nuclear metabolism, Sulfonamides pharmacology, ortho-Aminobenzoates pharmacology
- Abstract
Severe pulmonary hypertension (PH) is a disabling disease with high mortality, characterized by pulmonary vascular remodeling and right heart hypertrophy. In mice with PH induced by chronic hypoxia, we examined the acute and chronic effects of the soluble guanylate cyclase (sGC) activator HMR1766 on hemodynamics and pulmonary vascular remodeling. In isolated perfused mouse lungs from control animals, HMR1766 dose-dependently inhibited the pressor response of acute hypoxia. This dose-response curve was shifted leftward when the effects of HMR1766 were investigated in isolated lungs from chronic hypoxic animals for 21 days at 10% oxygen. Mice exposed for 21 or 35 days to chronic hypoxia developed PH, right heart hypertrophy, and pulmonary vascular remodeling. Treatment with HMR1766 (10 mg x kg(-1) x day(-1)), after full establishment of PH from day 21 to day 35, significantly reduced PH, as measured continuously by telemetry. In addition, right ventricular (RV) hypertrophy and structural remodeling of the lung vasculature were reduced. Pharmacological activation of oxidized sGC partially reverses hemodynamic and structural changes in chronic hypoxia-induced experimental PH.
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- 2009
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14. Heme oxygenase-2 and large-conductance Ca2+-activated K+ channels: lung vascular effects of hypoxia.
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Roth M, Rupp M, Hofmann S, Mittal M, Fuchs B, Sommer N, Parajuli N, Quanz K, Schubert D, Dony E, Schermuly RT, Ghofrani HA, Sausbier U, Rutschmann K, Wilhelm S, Seeger W, Ruth P, Grimminger F, Sausbier M, and Weissmann N
- Subjects
- Animals, Hypertension, Pulmonary pathology, Hypertension, Pulmonary physiopathology, Hypoxia pathology, In Vitro Techniques, Lung pathology, Mice, Microscopy, Fluorescence, Muscle, Smooth, Vascular pathology, Muscle, Smooth, Vascular physiopathology, Polymerase Chain Reaction, Pulmonary Alveoli pathology, Pulmonary Wedge Pressure physiology, RNA, Messenger genetics, Heme Oxygenase (Decyclizing) genetics, Heme Oxygenase (Decyclizing) physiology, Hypoxia physiopathology, Large-Conductance Calcium-Activated Potassium Channel alpha Subunits genetics, Large-Conductance Calcium-Activated Potassium Channel alpha Subunits physiology, Lung blood supply, Pulmonary Alveoli blood supply, Vasoconstriction physiology
- Abstract
Rationale: Hypoxic pulmonary vasoconstriction (HPV) is an important mechanism by which pulmonary gas exchange is optimized by the adaptation of blood flow to alveolar ventilation. In chronic hypoxia, in addition to HPV a vascular remodeling process leads to pulmonary hypertension. A complex of heme oxygenase-2 (HO-2) and the BK channel has been suggested as a universal oxygen sensor system., Objectives: We investigated whether this complex serves as an oxygen sensor for the vascular effects of alveolar hypoxia in the lung., Methods: The investigations were performed in chronically hypoxic mice, in isolated perfused and ventilated lungs, and on the cellular level, including HO-2- and BK-channel deficient mice., Measurements and Main Results: Immunohistochemical analysis of mouse lungs identified HO-2 mainly in pulmonary arteries, the bronchial epithelium, and alveolar epithelial cells. BK channel alpha-subunit (BKalpha) immunoreactivity was found primarily in the bronchial and vascular smooth muscle layer. Immunofluorescence staining and coimmunoprecipitation suggested only a weak complexation of HO-2 and BKalpha in pulmonary arterial smooth muscle cells. The strength of acute and sustained HPV, determined in isolated perfused and ventilated lungs, was not different among wild-type, HO-2-deficient, and BKalpha-deficient mice. Exposure of mice to 3 weeks of chronic hypoxia resulted in a slight down-regulation of HO-2 and no alteration in BKalpha expression. The degree of pulmonary hypertension that developed, quantified on the basis of right ventricular pressure, right-heart hypertrophy, and the degree of muscularization of precapillary pulmonary arteries, was not different among wild-type, HO-2-deficient, and BKalpha-deficient mice., Conclusions: It is demonstrated that neither deletion of HO-2 nor BK channels affect acute, sustained, and chronic vascular responses to alveolar hypoxia in the lung.
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- 2009
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15. Direct eicosanoid profiling of the hypoxic lung by comprehensive analysis via capillary liquid chromatography with dual online photodiode-array and tandem mass-spectrometric detection.
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Kiss L, Röder Y, Bier J, Weissmann N, Seeger W, and Grimminger F
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- Animals, Calibration, Eicosanoids chemistry, Molecular Structure, Rabbits, Solvents, Tandem Mass Spectrometry instrumentation, Chromatography, Liquid instrumentation, Chromatography, Liquid methods, Eicosanoids analysis, Hypoxia metabolism, Lung metabolism, Online Systems instrumentation, Tandem Mass Spectrometry methods
- Abstract
Eicosanoids are arachidonic acid-derived mediators, with partly contradictory, incompletely elucidated actions. Thus, epoxyeicosatrienoic acids (EETs) are controversially discussed as putative vasodilatative endothelium-derived hyperpolarizing factors in the cardiovascular compartment but reported as vasoconstrictors in the lung. Inconsistent findings concerning eicosanoid physiology may be because previous methods were lacking sensitivity, identification reliability, and/or have focused on special eicosanoid groups only, ignoring the overall mediator context, and thus limiting the correlation accuracy between autacoid formation and bioactivity profile. Therefore, we developed an approach which enables the simultaneous assessment of 44 eicosanoids, including all representatives of the arachidonic acid cascade, i.e., cytochrome P450, lipoxygenase, cyclooxygenase products, and free isoprostanes as in vivo markers of oxidative stress, in one 50-minute chromatographic run. The approach combines (i) source-specific sample extraction, (ii) rugged isocratic and high-sensitivity capillary liquid-chromatographic separation, and (iii) reliable dual online photodiode-array and electrospray ionization tandem mass-spectrometric identification and quantitation. High sensitivity with limits of quantification in the femtogram range was achieved by use of capillary columns with typical high peak efficiency, due to small inner diameters, and virtually complete substance transfer to the mass spectrometer, due to flow rates in the low microliter range, instead of large inner diameter columns with low chromatographic signal and only partial analyte transfer employed by previous methods. This expeditious, global and sensitive technique provides the prerequisite for new, accurate insights regarding the physiology of specific mediators, for example EETs, in the context of all relevant vasoactive autacoids under varying conditions of oxidative stress by direct comparison of all eicosanoid generation profiles. Indeed, application of comprehensive "eicoprofiling" to hypoxically ventilated rabbit lungs revealed at a glance the enhanced biosynthesis of free EETs in the overall mediator generation context, thus suggesting their hypothetical contribution to hypoxic pulmonary vasoconstriction.
- Published
- 2008
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16. Effect of sildenafil on hypoxia-induced changes in pulmonary circulation and right ventricular function.
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Reichenberger F, Kohstall MG, Seeger T, Olschewski H, Grimminger F, Seeger W, and Ghofrani HA
- Subjects
- Adult, Altitude, Double-Blind Method, Echocardiography, Doppler methods, Female, Follow-Up Studies, Humans, Male, Middle Aged, Mountaineering physiology, Purines therapeutic use, Sildenafil Citrate, Hypoxia drug therapy, Hypoxia physiopathology, Piperazines therapeutic use, Pulmonary Circulation drug effects, Sulfones therapeutic use, Vasodilator Agents therapeutic use, Ventricular Function, Right drug effects
- Abstract
Hypoxia leads to pulmonary vasoconstriction in healthy men. However, the consequences on right ventricular function are not known. The effects of hypoxia on systolic pulmonary artery pressure (sPAP) and right ventricular function index (TEI) were assessed by Doppler echocardiography. Fourteen members of a Mount Everest expedition were monitored during acute hypoxic challenge at sea level, environmental hypoxia exposure at altitudes of 3440 m and 5245 m and 2 weeks after return to sea level. Subjects received either placebo or 50mg sildenafil in a double-blind randomised cross-over design. Under normoxia at baseline, mean sPAP was 17.1(S.E.M. 1.3) mm Hg, and TEI was 0.13(0.004). Both increased during acute hypoxia: sPAP 29.6(2.6) mm Hg, and TEI 0.35(0.06) (each p<0.01). At 5245 m sPAP was 29.1(1.7) and TEI was 0.43(0.05) in the placebo group, while in the sildenafil group, both sPAP and TEI were reduced to 22(1.5) mm Hg and 0.23(0.03) (each p<0.005), respectively. We conclude that in healthy individuals, exposure to acute hypoxia and sojourns at high altitude result in a small but significant increase in sPAP accompanied by an impairment of right ventricular function. Sildenafil significantly decreases sPAP and improves right ventricular function.
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- 2007
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17. Hypoxia-induced pulmonary hypertension: different impact of iloprost, sildenafil, and nitric oxide.
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Weissmann N, Gerigk B, Kocer O, Nollen M, Hackemack S, Ghofrani HA, Schermuly RT, Butrous G, Schulz A, Roth M, Seeger W, and Grimminger F
- Subjects
- Analysis of Variance, Animals, Dose-Response Relationship, Drug, Drug Therapy, Combination, Hypertension, Pulmonary etiology, Hypertension, Pulmonary physiopathology, Iloprost administration & dosage, Iloprost pharmacology, Nitric Oxide administration & dosage, Nitric Oxide pharmacology, Piperazines administration & dosage, Piperazines pharmacology, Purines administration & dosage, Purines pharmacology, Rabbits, Sildenafil Citrate, Sulfones administration & dosage, Sulfones pharmacology, Treatment Outcome, Vasodilator Agents administration & dosage, Hypertension, Pulmonary drug therapy, Hypoxia complications, Vascular Resistance drug effects, Vasodilator Agents pharmacology
- Abstract
Objectives: Chronic alveolar hypoxia induces pulmonary hypertension, evident from elevated pulmonary artery pressure (PAP), pulmonary vascular resistance, right ventricular hypertrophy (RVH), and increased muscularization of the pulmonary vasculature. Additionally, the vasoconstrictor response to acute hypoxia (HPV) may be reduced in the remodeled vasculature. However, no direct comparison of different treatments on the various parameters characterizing pulmonary hypertension has been performed yet. Against this background, we compared the effects of inhaled NO, infused iloprost, a stable prostacyclin analogue, and oral sildenafil, a phosphodiesterase 5 inhibitor, on hypoxia-induced pulmonary hypertension., Methods: Exposure of rabbits to chronic hypoxia (FiO(2)=0.10) for 42 days. Treatment with infused iloprost, oral sildenafil, and inhaled nitric oxide., Results: We quantified PAP, pulmonary vascular resistance, RVH, vascular remodeling, vasoreactivity, and the strength of HPV. Chronic hypoxia resulted in an increase in (a) the right ventricle/(left ventricle+septum) ratio from 0.26+/-0.01 to 0.44+/-0.01, (b) PAP, and (c) the degree of muscularization from 14.0+/-4.0% to 43.5+/-5.3%. Treatment with iloprost and sildenafil, but not with NO, prevented the increase in muscularization. In contrast, RVH was strongly inhibited by sildenafil, whereas NO had some minor, and iloprost had no effect. Only iloprost reduced PAP compared to NO and sildenafil. The downregulation of HPV was abrogated only by NO., Conclusion: We demonstrated (a) that the parameters characterizing hypoxia-induced pulmonary hypertension are not functionally linked, (b) that the downregulation of HPV under chronic hypoxia can be prevented by inhaled NO but not by sildenafil and iloprost, and (c) that iloprost is particularly effective in preventing vascular remodeling and sildenafil in preventing RVH.
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- 2007
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18. Hypoxia-dependent regulation of nonphagocytic NADPH oxidase subunit NOX4 in the pulmonary vasculature.
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Mittal M, Roth M, König P, Hofmann S, Dony E, Goyal P, Selbitz AC, Schermuly RT, Ghofrani HA, Kwapiszewska G, Kummer W, Klepetko W, Hoda MA, Fink L, Hänze J, Seeger W, Grimminger F, Schmidt HH, and Weissmann N
- Subjects
- Animals, Cell Division, Cells, Cultured drug effects, Cells, Cultured enzymology, Chronic Disease, Drug Design, Endoplasmic Reticulum enzymology, Enzyme Induction, Female, Humans, Hypertension, Pulmonary drug therapy, Hypertension, Pulmonary etiology, Hypertension, Pulmonary physiopathology, Hypertrophy, Hypoxia complications, Hypoxia physiopathology, Hypoxia-Inducible Factor 1, alpha Subunit metabolism, Lung blood supply, Male, Membrane Glycoproteins analysis, Myocytes, Smooth Muscle enzymology, Myocytes, Smooth Muscle pathology, NADPH Oxidase 2, NADPH Oxidase 4, NADPH Oxidases analysis, NADPH Oxidases biosynthesis, NADPH Oxidases genetics, Nitric Oxide physiology, Organ Specificity, Oxygen metabolism, Oxygen pharmacology, Protein Subunits, Pulmonary Artery cytology, Pulmonary Artery enzymology, RNA Interference, RNA, Messenger biosynthesis, RNA, Small Interfering pharmacology, Superoxides metabolism, Transforming Growth Factor beta1 physiology, Tunica Media enzymology, Tunica Media pathology, Hypertension, Pulmonary enzymology, Hypoxia enzymology, NADPH Oxidases physiology
- Abstract
Nonphagocytic NADPH oxidases have recently been suggested to play a major role in the regulation of physiological and pathophysiological processes, in particular, hypertrophy, remodeling, and angiogenesis in the systemic circulation. Moreover, NADPH oxidases have been suggested to serve as oxygen sensors in the lung. Chronic hypoxia induces vascular remodeling with medial hypertrophy leading to the development of pulmonary hypertension. We screened lung tissue for the expression of NADPH oxidase subunits. NOX1, NOXA1, NOXO1, p22phox, p47phox, p40phox, p67phox, NOX2, and NOX4 were present in mouse lung tissue. Comparing mice maintained for 21 days under hypoxic (10% O(2)) or normoxic (21% O(2)) conditions, an upregulation exclusively of NOX4 mRNA was observed under hypoxia in homogenized lung tissue, concomitant with increased levels in microdissected pulmonary arterial vessels. In situ hybridization and immunohistological staining for NOX4 in mouse lungs revealed a localization of NOX4 mRNA and protein predominantly in the media of small pulmonary arteries, with increased labeling intensities after chronic exposure to hypoxia. In isolated pulmonary arterial smooth muscle cells (PASMCs), NOX4 was localized primarily to the perinuclear space and its expression levels were increased after exposure to hypoxia. Treatment of PASMCs with siRNA directed against NOX4 decreased NOX4 mRNA levels and reduced PASMC proliferation as well as generation of reactive oxygen species. In lungs from patients with idiopathic pulmonary arterial hypertension (IPAH), expression levels of NOX4, which was localized in the vessel media, were 2.5-fold upregulated. These results support an important role for NOX4 in the vascular remodeling associated with development of pulmonary hypertension.
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- 2007
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19. Classical transient receptor potential channel 6 (TRPC6) is essential for hypoxic pulmonary vasoconstriction and alveolar gas exchange.
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Weissmann N, Dietrich A, Fuchs B, Kalwa H, Ay M, Dumitrascu R, Olschewski A, Storch U, Mederos y Schnitzler M, Ghofrani HA, Schermuly RT, Pinkenburg O, Seeger W, Grimminger F, and Gudermann T
- Subjects
- Acute Disease, Animals, Cations chemistry, Cations metabolism, Diglycerides metabolism, Gene Expression, Hypoxia genetics, Lung Diseases genetics, Membrane Potentials, Mice, Mice, Knockout, TRPC Cation Channels deficiency, TRPC Cation Channels genetics, TRPC6 Cation Channel, Aorta metabolism, Hypoxia metabolism, Lung Diseases metabolism, Pulmonary Gas Exchange, TRPC Cation Channels metabolism, Vasoconstriction
- Abstract
Regional alveolar hypoxia causes local vasoconstriction in the lung, shifting blood flow from hypoxic to normoxic areas, thereby maintaining gas exchange. This mechanism is known as hypoxic pulmonary vasoconstriction (HPV). Disturbances in HPV can cause life-threatening hypoxemia whereas chronic hypoxia triggers lung vascular remodeling and pulmonary hypertension. The signaling cascade of this vitally important mechanism is still unresolved. Using transient receptor potential channel 6 (TRPC6)-deficient mice, we show that this channel is a key regulator of acute HPV as this regulatory mechanism was absent in TRPC6(-/-) mice whereas the pulmonary vasoconstrictor response to the thromboxane mimetic U46619 was unchanged. Accordingly, induction of regional hypoventilation resulted in severe arterial hypoxemia in TRPC6(-/-) but not in WT mice. This effect was mirrored by a lack of hypoxia-induced cation influx and currents in smooth-muscle cells from precapillary pulmonary arteries (PASMC) of TRPC6(-/-) mice. In both WT and TRPC6(-/-) PASMC hypoxia caused diacylglycerol (DAG) accumulation. DAG seems to exert its action via TRPC6, as DAG kinase inhibition provoked a cation influx only in WT but not in TRPC6(-/-) PASMC. Notably, chronic hypoxia-induced pulmonary hypertension was independent of TRPC6 activity. We conclude that TRPC6 plays a unique and indispensable role in acute hypoxic pulmonary vasoconstriction. Manipulation of TRPC6 function may thus offer a therapeutic strategy for the control of pulmonary hemodynamics and gas exchange.
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- 2006
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20. Hypoxia- and non-hypoxia-related pulmonary hypertension - established and new therapies.
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Ghofrani HA, Voswinckel R, Reichenberger F, Weissmann N, Schermuly RT, Seeger W, and Grimminger F
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- Animals, Antihypertensive Agents therapeutic use, Humans, Hypertension, Pulmonary drug therapy, Hypoxia drug therapy, Lung Diseases, Interstitial complications, Lung Diseases, Interstitial drug therapy, Pulmonary Disease, Chronic Obstructive complications, Pulmonary Disease, Chronic Obstructive drug therapy, Thrombosis complications, Thrombosis drug therapy, Vasodilator Agents therapeutic use, Hypertension, Pulmonary etiology, Hypoxia complications
- Abstract
Pulmonary hypertension can occur as an isolated disease affecting the lung vessels only, in association with underlying hypoxic lung disorders, or due to chronic thromboembolic disease. Pulmonary hypertension caused by pulmonary venous congestion will not be focused on in this review. Regardless of the underlying disease, chronic cor pulmonale is associated with progressive clinical deterioration and a poor prognosis in most cases. The aim of specific therapies for pulmonary hypertension is to reduce pulmonary vascular resistance and thereby improve right ventricular function. Currently, three classes of drugs (prostanoids, endothelin receptor antagonists, and phosphodiesterase-5 inhibitors) are approved for the treatment of pulmonary arterial hypertension (PAH) in a defined patient population (group I according to the recent WHO classification). However, these medications may also lower pulmonary vascular resistance in patients with associated lung diseases (e.g. chronic obstructive pulmonary disease or lung fibrosis) and significant pulmonary hypertension, for whom these drugs are not yet approved. As non-selective vasodilators may induce gas-exchange disturbances, which preclude their long term use in these patients, such substances should be avoided in the hypoxemic patient. In this article we provide an update of the current understanding of hypoxia- and non-hypoxia-related pulmonary hypertension, addressing both the pathophysiological understanding of different disease aetiologies as well as the therapeutic options currently available.
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- 2006
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21. Hypoxia in lung vascular biology and disease.
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Weissmann N, Grimminger F, and Seeger W
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- Adaptation, Physiological, Animals, Humans, Hypertension, Pulmonary physiopathology, Lung physiopathology, Pulmonary Gas Exchange, Vasoconstriction physiology, Blood-Air Barrier, Hypoxia metabolism, Lung physiology, Pulmonary Circulation
- Published
- 2006
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22. Oxygen sensors in hypoxic pulmonary vasoconstriction.
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Weissmann N, Sommer N, Schermuly RT, Ghofrani HA, Seeger W, and Grimminger F
- Subjects
- Adenosine Triphosphate metabolism, Animals, Calcium metabolism, Cytochrome P-450 Enzyme System metabolism, Heme Oxygenase (Decyclizing) metabolism, Humans, Hypertension, Pulmonary metabolism, Mitochondria metabolism, Reactive Oxygen Species metabolism, Vasoconstriction physiology, Hypoxia metabolism, Lung metabolism, Oxygen metabolism
- Abstract
Hypoxic pulmonary vasoconstriction (HPV) is an essential mechanism adapting lung perfusion to regional ventilation. Perturbations to HPV, such as those occurring in pneumonia, acute respiratory distress syndrome and liver failure, can result in arterial hypoxemia. Under conditions of general hypoxia, HPV increases pulmonary vascular resistance and thus causes acute pulmonary hypertension. Despite intensive research, the underlying mechanisms of HPV have not been fully elucidated. Deciphering signalling pathways that result in HPV could suggest novel approaches to address a failure of HPV, as well as for the treatment of pulmonary hypertension associated with HPV. Within this context, this review focuses on current concepts in the oxygen sensing mechanisms that underlie HPV.
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- 2006
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23. Impact of mitochondria and NADPH oxidases on acute and sustained hypoxic pulmonary vasoconstriction.
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Weissmann N, Zeller S, Schäfer RU, Turowski C, Ay M, Quanz K, Ghofrani HA, Schermuly RT, Fink L, Seeger W, and Grimminger F
- Subjects
- 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid pharmacology, Animals, Electron Transport Complex I physiology, Electron Transport Complex II physiology, Electron Transport Complex III physiology, Electron Transport Complex IV physiology, Female, Hypoxia physiopathology, In Vitro Techniques, Male, Membrane Glycoproteins genetics, Mice, Mice, Inbred C57BL, NADPH Oxidase 2, NADPH Oxidases antagonists & inhibitors, NADPH Oxidases genetics, Protein Isoforms antagonists & inhibitors, Protein Isoforms physiology, Pulmonary Gas Exchange, Rabbits, Superoxides metabolism, Vasoconstrictor Agents pharmacology, Hypoxia metabolism, Lung blood supply, Mitochondria physiology, NADPH Oxidases physiology, Oxygen physiology, Vasoconstriction drug effects
- Abstract
Hypoxic pulmonary vasoconstriction (HPV) matches lung perfusion with ventilation to optimize pulmonary gas exchange. However, it remains unclear whether acute HPV (occurring within seconds) and the vasoconstrictor response to sustained alveolar hypoxia (developing over several hours) are triggered by identical mechanisms. We investigated the effect of mitochondrial and NADPH oxidase inhibitors on both phases of HPV in intact rabbit lungs. These studies revealed that the sustained HPV is largely dependent on mitochondrial complex I and totally dependent on complex IV, whereas NADPH oxidase dependence was only observed for acute HPV. These findings were reinforced by an alternative approach employing lungs from mice deficient in the NADPH oxidase subunit p 47(phox). In these mice (which lack a subunit suggested to be important for the function of most NADPH oxidase isoforms), but not in gp 91(phox)-deficient mice (which represent only one isoform of NADPH oxidases), acute HPV was significantly reduced, while non-hypoxia-induced vasoconstrictions elicited by the thromboxane mimetic U46619 were not affected. We concluded that the acute phase and the sustained phase of HPV are differentially regulated, with NADPH oxidase activity predominating in the acute phase, while a strong dependence on mitochondrial participation was observed for the second phase.
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- 2006
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24. Hypoxic pulmonary vasoconstriction--triggered by an increase in reactive oxygen species?
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Weissmann N, Schermuly RT, Ghofrani HA, Hänze J, Goyal P, Grimminger F, and Seeger W
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- Animals, Pulmonary Artery physiopathology, Rabbits, Reactive Oxygen Species antagonists & inhibitors, Hypoxia metabolism, Hypoxia physiopathology, Pulmonary Artery metabolism, Reactive Oxygen Species metabolism, Vasoconstriction physiology
- Abstract
Hypoxic pulmonary vasoconstriction (HPV) is an essential mechanism of the lung that matches perfusion to ventilation in order to optimize pulmonary gas exchange. Despite intensive research, the underlying mechanism has not yet been fully elucidated. Reactive oxygen species (ROS) have been proposed as key mediators of HPV. However, there is ongoing discussion as to whether ROS really contribute to HPV regulation and if so, whether an increase or a decrease in ROS occurs during alveolar hypoxia. In this overview, we summarize our data that have led us to conclude that alveolar hypoxia induces an increase in superoxide and subsequently H2O2, and thus elicits HPV. This conclusion is drawn from investigations employing various inhibitors that interfere with ROS in isolated buffer-perfused rabbit lungs challenged with 10-minute periods of alveolar hypoxia. Targeting possible sources of a hypoxia-induced increase in ROS, our data are only partially in accordance with the hypothesis that mitochondria are the hypoxia-dependent ROS generators, and suggest NADPH oxidases as an alternative source. From measurements of intracellular and exhaled H2O2, we hypothesize that total lung ROS release is reduced in alveolar hypoxia, but that in specialized cells or sub-cellular structures an increased ROS release may occur, triggering HPV.
- Published
- 2006
25. Impact of HIF-1alpha and HIF-2alpha on proliferation and migration of human pulmonary artery fibroblasts in hypoxia.
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Eul B, Rose F, Krick S, Savai R, Goyal P, Klepetko W, Grimminger F, Weissmann N, Seeger W, and Hänze J
- Subjects
- Actins metabolism, Apoptosis, Basic Helix-Loop-Helix Transcription Factors, Cell Cycle, Cell Proliferation, Cells, Cultured, Humans, Hypoxia-Inducible Factor 1, alpha Subunit genetics, Oxygen metabolism, RNA Interference, Transcription Factors genetics, Cell Movement, Fibroblasts cytology, Fibroblasts metabolism, Hypoxia metabolism, Hypoxia-Inducible Factor 1, alpha Subunit metabolism, Pulmonary Artery cytology, Transcription Factors metabolism
- Abstract
Proliferation of adventitial fibroblasts of small intrapulmonary arteries (FBPA) has been disclosed as an early event in the development of pulmonary hypertension and cor pulmonale in response to hypoxia. We investigated the role of hypoxia-inducible transcription factors (HIF) in human FBPA exposed to hypoxia. Primary cultures of FBPA displayed a strong mitogenic response to 24 h hypoxia, whereas the rate of apoptosis was significantly suppressed. In addition, the migration of FBPA was strongly increased under hypoxic conditions but not the expression of alpha-smooth muscle actin. Hypoxia induced a marked up-regulation (protein level) of both HIF-1alpha and HIF-2alpha, alongside with nuclear translocation of these transcription factors. Specific inhibition of either HIF-1alpha or HIF-2alpha was achieved by RNA interference technology, as proven by HIF-1alpha and HIF-2alpha mRNA and protein analysis and expression analysis of HIF downstream target genes. With the use of this approach, the hypoxia-induced proliferative response of the FBPA was found to be solely HIF-2alpha dependent, whereas the migratory response was significantly reduced by both HIF-1alpha and HIF-2alpha interference. In conclusion, HIF up-regulation is essential for hypoxic cellular responses in human pulmonary artery adventitial fibroblasts such as proliferation and migration, mimicking the pulmonary hypertensive phenotype in vivo. Differential HIF subtype dependency was noted, with HIF-2alpha playing a predominant role, which may offer future intervention strategies.
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- 2006
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26. Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertension.
- Author
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Kwapiszewska G, Wilhelm J, Wolff S, Laumanns I, Koenig IR, Ziegler A, Seeger W, Bohle RM, Weissmann N, and Fink L
- Subjects
- Animals, Gene Expression Profiling, Hypertension, Pulmonary etiology, Hypoxia complications, Laser Therapy, Male, Mice, Mice, Inbred BALB C, Microdissection, Cytokines metabolism, Hypertension, Pulmonary metabolism, Hypertension, Pulmonary pathology, Hypoxia metabolism, Hypoxia pathology, Pulmonary Artery metabolism, Pulmonary Artery pathology
- Abstract
Background: Chronic hypoxia influences gene expression in the lung resulting in pulmonary hypertension and vascular remodelling. For specific investigation of the vascular compartment, laser-microdissection of intrapulmonary arteries was combined with array profiling., Methods and Results: Analysis was performed on mice subjected to 1, 7 and 21 days of hypoxia (FiO2 = 0.1) using nylon filters (1176 spots). Changes in the expression of 29, 38, and 42 genes were observed at day 1, 7, and 21, respectively. Genes were grouped into 5 different classes based on their time course of response. Gene regulation obtained by array analysis was confirmed by real-time PCR. Additionally, the expression of the growth mediators PDGF-B, TGF-beta, TSP-1, SRF, FGF-2, TIE-2 receptor, and VEGF-R1 were determined by real-time PCR. At day 1, transcription modulators and ion-related proteins were predominantly regulated. However, at day 7 and 21 differential expression of matrix producing and degrading genes was observed, indicating ongoing structural alterations. Among the 21 genes upregulated at day 1, 15 genes were identified carrying potential hypoxia response elements (HREs) for hypoxia-induced transcription factors. Three differentially expressed genes (S100A4, CD36 and FKBP1a) were examined by immunohistochemistry confirming the regulation on protein level. While FKBP1a was restricted to the vessel adventitia, S100A4 and CD36 were localised in the vascular tunica media., Conclusion: Laser-microdissection and array profiling has revealed several new genes involved in lung vascular remodelling in response to hypoxia. Immunohistochemistry confirmed regulation of three proteins and specified their localisation in vascular smooth muscle cells and fibroblasts indicating involvement of different cells types in the remodelling process. The approach allows deeper insight into hypoxic regulatory pathways specifically in the vascular compartment of this complex organ.
- Published
- 2005
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27. Congenital erythropoietin over-expression causes "anti-pulmonary hypertensive" structural and functional changes in mice, both in normoxia and hypoxia.
- Author
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Weissmann N, Manz D, Buchspies D, Keller S, Mehling T, Voswinckel R, Quanz K, Ghofrani HA, Schermuly RT, Fink L, Seeger W, Gassmann M, and Grimminger F
- Subjects
- 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid pharmacology, Animals, Aspirin pharmacology, Blood Pressure, Blood Vessels pathology, Blood Viscosity, Cyclooxygenase Inhibitors pharmacology, Erythropoietin blood, Erythropoietin genetics, Hematocrit, Hypertension, Pulmonary blood, Hypertension, Pulmonary etiology, Hypertension, Pulmonary physiopathology, Hypertrophy, Left Ventricular physiopathology, Hypertrophy, Right Ventricular physiopathology, Lung pathology, Lung physiopathology, Mice, Mice, Inbred C57BL, Mice, Transgenic, Muscle, Smooth, Vascular pathology, Nitric Oxide Synthase antagonists & inhibitors, Up-Regulation, Vascular Resistance drug effects, Vasoconstrictor Agents pharmacology, omega-N-Methylarginine pharmacology, Erythropoietin physiology, Hypertension, Pulmonary prevention & control, Hypoxia physiopathology, Lung blood supply, Vasoconstriction drug effects
- Abstract
Acute alveolar hypoxia causes pulmonary vasoconstriction that matches lung perfusion to ventilation to optimize gas exchange. Chronic alveolar hypoxia induces pulmonary hypertension, characterized by increased muscularization of the pulmonary vasculature and right ventricular hypertrophy. Elevated erythropoietin (EPO) plasma levels increase hematocrit and blood viscosity and may affect structure and function of the pulmonary circulation. To differentiate between the direct effects of hypoxia and those linked to a hypoxia-induced increase in EPO/hematocrit levels, we investigated the lung vasculature in transgenic mice constitutively over-expressing EPO (termed tg6) upon exposure to normoxia and chronic hypoxia. Despite increased hematocrit levels (approximately 0.86),tg6 mice kept in normoxia did not develop selective right ventricular hypertrophy. The portion of vessels with a diameter of 51-95 microm and >155 microm was increased whereas the portion of small vessels (30-50 microm) was decreased. Pulmonary vascular resistance and the strength of hypoxic vasoconstriction measured in isolated perfused lungs were decreased. Vasoconstrictions induced by the thromboxane mimetic U46619 tended to be reduced. After chronic hypoxia (FiO2 = 0.10, 21 days), vascular resistance and vasoconstrictor responses to acute hypoxia and U46619 were reduced in tg6 mice compared to wildtype controls. Chronic hypoxia increased the degree of pulmonary vascular muscularization in wildtype but not in tg6 mice that already exhibited less muscularization in normoxia. In conclusion, congenital over-expression of EPO exerts an "anti-pulmonary hypertensive" effect, both structurally and functionally, particularly obvious upon chronic hypoxia.
- Published
- 2005
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28. Role of hypoxia-inducible factor-1alpha in hypoxia-induced apoptosis of primary alveolar epithelial type II cells.
- Author
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Krick S, Eul BG, Hänze J, Savai R, Grimminger F, Seeger W, and Rose F
- Subjects
- Animals, Caspase Inhibitors, Caspases metabolism, Cell Cycle physiology, Cell Proliferation, Cells, Cultured, DNA-Binding Proteins genetics, Homeostasis, Hypoxia-Inducible Factor 1, Hypoxia-Inducible Factor 1, alpha Subunit, Male, Membrane Proteins metabolism, Nuclear Proteins genetics, Proto-Oncogene Proteins metabolism, RNA, Small Interfering metabolism, Rats, Rats, Sprague-Dawley, Transcription Factors genetics, Tumor Suppressor Proteins metabolism, Apoptosis physiology, DNA-Binding Proteins metabolism, Hypoxia, Nuclear Proteins metabolism, Pulmonary Alveoli cytology, Pulmonary Alveoli metabolism, Respiratory Mucosa cytology, Transcription Factors metabolism
- Abstract
Hypoxia affects alveolar homeostasis and may induce epithelial injury, which has been implicated in lung diseases such as fibrosis. The underlying cellular and molecular mechanisms are, however, largely unknown. Primary rat alveolar epithelial type II cells (ATII) exposed to graded hypoxia for 24 and 48 h caused a dose-dependent induction of cell cycle arrest and suppression of proliferation, which were comparable to the effects of angiotensin II, a potent inducer of ATII cell death. Hypoxia-induced changes in ATII homeostasis are thought to proceed primarily via activation of hypoxia inducible-factor (HIF)-1alpha, because hypoxia increased HIF-1alpha protein expression, nuclear translocation, and transactivation of its specific DNA binding domain, the hypoxia responsive element (HRE). Under hypoxic conditions, expression of the proapoptotic protein Bnip3L, which belongs to the Bcl 2 family and is known to be one of the HIF-1-dependent target genes, was upregulated. Suppression of HIF-1alpha or Bnip-3L with small interfering RNA (siRNA) fully blocked the hypoxia-induced apoptosis and Bnip3L expression. In line with these data, overexpression of HIF-1alpha by transient transfection enhanced the hypoxia-induced apoptosis. Thus, we conclude that hypoxia suppresses alveolar epithelial cell proliferation and enhances ATII apoptosis through activation of the HIF-1alpha/HRE axis and a mechanism that involves Bnip3L. Targeting HIF-1alpha may represent a new strategy that could impede the alveolar denudation that is observed in several lung diseases.
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- 2005
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29. Hypoxia-driven proliferation of human pulmonary artery fibroblasts: cross-talk between HIF-1alpha and an autocrine angiotensin system.
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Krick S, Hänze J, Eul B, Savai R, Seay U, Grimminger F, Lohmeyer J, Klepetko W, Seeger W, and Rose F
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- Angiotensin II pharmacology, Angiotensin II Type 1 Receptor Blockers pharmacology, Angiotensin-Converting Enzyme Inhibitors pharmacology, Apoptosis drug effects, Cell Cycle drug effects, Cell Division drug effects, Cells, Cultured, Connective Tissue pathology, Feedback, Physiological, Gene Expression Regulation drug effects, Humans, Hypoxia-Inducible Factor 1, alpha Subunit genetics, Peptidyl-Dipeptidase A genetics, Phosphoinositide-3 Kinase Inhibitors, RNA, Small Interfering pharmacology, Receptor, Angiotensin, Type 1 genetics, Response Elements genetics, Signal Transduction, Angiotensins physiology, Cell Division physiology, Fibroblasts pathology, Hypoxia pathology, Hypoxia-Inducible Factor 1, alpha Subunit physiology, Pulmonary Artery pathology
- Abstract
Pulmonary artery adventitial fibroblasts (FBPA) may play a central role in lung vascular remodeling under conditions of hypoxia and inflammation, the result being pulmonary hypertension and cor pulmonale. In cultured human FBPA, both angiotensin II (Ang II) and hypoxia promoted cell cycle progression and cell proliferation and suppressed apoptosis. These effects were further enhanced when both stimuli were applied simultaneously. Hypoxia elevated the expression of hypoxia-inducible factor 1alpha (HIF-1alpha) and increased the expression of genes regulated by the hypoxia-responsive element (HRE). Up-regulation of both angiotensin-converting enzyme (ACE) and Ang II receptor type 1 (AT1) was also observed. Exogenous Ang II further increased HIF/HRE-dependent signaling in FBPA, whereas suppression of the autocrine ACE-Ang II-AT1 loop with inhibitors of ACE, AT1, and phosphatidylinositol 3-kinase (PI3K) reduced the proliferative response to both hypoxia and exogenous Ang II. Overexpression of HIF-1alpha by transient transfection caused the same proliferative effect and up-regulation of AT1 expression that were observed under hypoxic conditions. In contrast, small interfering RNA targeting HIF-1alpha inhibited hypoxia-induced ACE and AT1 expression. Our studies indicate that the ACE-Ang II-AT1 system serves as a positive feedback loop and fosters FBPA proliferation under hypoxic conditions, with the PI3K-HIF-HRE axis as the central effector pathway. This pathway may thus facilitate vascular remodeling under hypoxic conditions.
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- 2005
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30. Sildenafil increased exercise capacity during hypoxia at low altitudes and at Mount Everest base camp: a randomized, double-blind, placebo-controlled crossover trial.
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Ghofrani HA, Reichenberger F, Kohstall MG, Mrosek EH, Seeger T, Olschewski H, Seeger W, and Grimminger F
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- Blood Pressure drug effects, Cardiac Output drug effects, Cross-Over Studies, Double-Blind Method, Female, Humans, Hypertension, Pulmonary etiology, Hypertension, Pulmonary physiopathology, Hypoxia complications, Hypoxia physiopathology, Male, Oxygen blood, Placebos, Pulmonary Artery physiology, Purines, Sildenafil Citrate, Sulfones, 3',5'-Cyclic-GMP Phosphodiesterases antagonists & inhibitors, Altitude, Exercise Tolerance drug effects, Hypertension, Pulmonary drug therapy, Hypoxia drug therapy, Phosphodiesterase Inhibitors therapeutic use, Piperazines therapeutic use, Vasodilator Agents therapeutic use
- Abstract
Background: Alveolar hypoxia causes pulmonary hypertension and enhanced right ventricular afterload, which may impair exercise tolerance. The phosphodiesterase-5 inhibitor sildenafil has been reported to cause pulmonary vasodilatation., Objective: To investigate the effects of sildenafil on exercise capacity under conditions of hypoxic pulmonary hypertension., Design: Randomized, double-blind, placebo-controlled crossover study., Setting: University Hospital Giessen, Giessen, Germany, and the base camp on Mount Everest., Participants: 14 healthy mountaineers and trekkers., Measurements: Systolic pulmonary artery pressure, cardiac output, and peripheral arterial oxygen saturation at rest and during assessment of maximum exercise capacity on cycle ergometry 1) while breathing a hypoxic gas mixture with 10% fraction of inspired oxygen at low altitude (Giessen) and 2) at high altitude (the Mount Everest base camp)., Intervention: Oral sildenafil, 50 mg, or placebo., Results: At low altitude, acute hypoxia reduced arterial oxygen saturation to 72.0% (95% CI, 66.5% to 77.5%) at rest and 60.8% (CI, 56.0% to 64.5%) at maximum exercise capacity. Systolic pulmonary artery pressure increased from 30.5 mm Hg (CI, 26.0 to 35.0 mm Hg) at rest to 42.9 mm Hg (CI, 35.6 to 53.5 mm Hg) during exercise in participants taking placebo. Sildenafil, 50 mg, significantly increased arterial oxygen saturation during exercise (P = 0.005) and reduced systolic pulmonary artery pressure at rest (P < 0.001) and during exercise (P = 0.031). Of note, sildenafil increased maximum workload (172.5 W [CI, 147.5 to 200.0 W]) vs. 130.6 W [CI, 108.8 to 150.0 W]); P < 0.001) and maximum cardiac output (P < 0.001) compared with placebo. At high altitude, sildenafil had no effect on arterial oxygen saturation at rest and during exercise compared with placebo. However, sildenafil reduced systolic pulmonary artery pressure at rest (P = 0.003) and during exercise (P = 0.021) and increased maximum workload (P = 0.002) and cardiac output (P = 0.015). At high altitude, sildenafil exacerbated existing headache in 2 participants., Limitations: The study did not examine the effects of sildenafil on normoxic exercise tolerance., Conclusions: Sildenafil reduces hypoxic pulmonary hypertension at rest and during exercise while maintaining gas exchange and systemic blood pressure. To the authors' knowledge, sildenafil is the first drug shown to increase exercise capacity during severe hypoxia both at sea level and at high altitude.
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- 2004
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31. Basic features of hypoxic pulmonary vasoconstriction in mice.
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Weissmann N, Akkayagil E, Quanz K, Schermuly RT, Ghofrani HA, Fink L, Hänze J, Rose F, Seeger W, and Grimminger F
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- 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid pharmacology, Amylopectin metabolism, Animals, Aspirin pharmacology, Blood Pressure drug effects, Drug Interactions, Enzyme Inhibitors pharmacology, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Pulmonary Artery drug effects, Pulmonary Circulation drug effects, Pulmonary Gas Exchange physiology, Species Specificity, Tidal Volume physiology, Vasoconstriction drug effects, Vasoconstrictor Agents pharmacology, omega-N-Methylarginine pharmacology, Amylopectin analogs & derivatives, Blood Pressure physiology, Hypoxia physiopathology, Oxygen metabolism, Pulmonary Artery physiopathology, Vasoconstriction physiology
- Abstract
Hypoxic pulmonary vasoconstriction (HPV) matches lung perfusion with ventilation which tends to optimize pulmonary gas exchange. Investigations using genetically engineered mice represent a promising approach to understand the underlying mechanisms. Our goal was to characterize basic features of HPV in the isolated buffer-perfused and ventilated mouse lung system. HPV was reproducible for several hours when ventilating the lungs with 1% O2 (10 min) alternated with normoxic ventilation periods (21% O2, 15 min). HPV was well elicitable and most constant using Krebs-Henseleit buffer with the addition of hydroxyethylamylopectin as an oncotic agent. Inhibition of both lung NO and prostanoid formation amplified HPV in an over-additive fashion. HPV was higher in BALB/c mive as compared to C57BL/6 mice, and was approximately threefold enhanced under positive pressure ventilation as compared to negative pressure ventilation. A three hour hypoxic ventilation period resulted in a biphasic vasoconstrictor response with loss of posthypoxic vasodilatation. In summary, we have characterised HPV and established an experimental set-up optimized for investigation of the basic mechanisms of HPV in mice.
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- 2004
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32. Increase in alveolar antioxidant levels in hyperoxic and anoxic ventilated rabbit lungs during ischemia.
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Schmidt R, Schäfer C, Luboeinski T, Löckinger A, Hermle G, Grimminger F, Seeger W, Ghofrani A, Schütte H, and Günther A
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- Animals, Bronchoalveolar Lavage Fluid chemistry, L-Lactate Dehydrogenase metabolism, Oxidation-Reduction, Plasmalogens metabolism, Pulmonary Surfactants metabolism, Rabbits, Uric Acid metabolism, Vitamins metabolism, Antioxidants metabolism, Hyperoxia metabolism, Hypoxia metabolism, Ischemia metabolism, Lung metabolism, Pulmonary Ventilation
- Abstract
Increases in free radicals are believed to play a central role in the development of pulmonary ischemia/reperfusion (I-R) injury, leading to microvascular leakage and deterioration of pulmonary surfactant. Continued ventilation during ischemia offers significant protection against I-R injury, but the impact of alveolar oxygen supply both on lung injury and on radical generation is still unclear. We investigated the influence of hyperoxic (95% O2) and anoxic (0% O2) ventilation during ischemia on alveolar antioxidant status and surfactant properties in isolated rabbit lungs. Normoxic and hyperoxic ventilated, buffer-perfused lungs (n = 5 or 6) and native lungs (n = 6) served as controls. As compared with controls, biophysical and biochemical surfactant properties were not altered in anoxic as well as hyperoxic ventilated ischemic (2, 3, and 4 h) lungs. Assessment of several antioxidants (reduced glutathione (GSH), alpha-tocopherol (vitamin E), retinol (vitamin A), ascorbic acid (vitamin C), uric acid, and plasmalogens (1-O-alkenyl-2-acyl-phospholipids)) in bronchoalveolar lavage fluid (BALF) revealed a significant increase in antioxidant compounds under anoxic and hyperoxic ventilation, with maximum levels occuring after 3 h of ischemia. For example, GSH increased to 5.1 +/- 0.8 microM (mean +/- SE, p <.001) after 3 h of anoxic ventilated ischemia and to 2.7 +/- 0.2 microM (p <.01) after hyperoxic ventilated ischemia compared with native controls (1.3 +/- 0.2 microM), but did not significantly change under anoxic and hyperoxic ventilation alone. In parallel, under ischemic conditions, oxidized glutathione (GSSG) increased during hyperoxic (3 h: 0.81 +/- 0.04 microM, p <.001), but remained unchanged during anoxic (3 h: 0.31 +/- 0.04 microM) ventilation compared with native controls (0.22 +/- 0.02 microM), whereas F2-isoprostanes were elevated under both hyperoxic (3 h: 63 +/- 15 pM, p <.01) and anoxic (3 h: 50 +/- 9 pM, p <.01) ventilation compared with native controls (16 +/- 4 pM). We conclude that oxidative stress is increased in the lung alveolar lining layer during ischemia, during both anoxic and hyperoxic ventilation. This is paralleled by an increase rather than a decrease in alveolar antioxidant levels, suggested to reflect an adaptive response to oxidative stress during ischemia.
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- 2004
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33. RNA interference for HIF-1alpha inhibits its downstream signalling and affects cellular proliferation.
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Hänze J, Eul BG, Savai R, Krick S, Goyal P, Grimminger F, Seeger W, and Rose F
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- Cell Division physiology, Cell Line, Humans, Hypoxia genetics, Hypoxia-Inducible Factor 1, alpha Subunit, Respiratory Mucosa cytology, Down-Regulation genetics, Hypoxia metabolism, RNA Interference physiology, Respiratory Mucosa growth & development, Respiratory Mucosa metabolism, Signal Transduction physiology, Transcription Factors genetics, Transcription Factors metabolism
- Abstract
Transcription factor HIF-1 is a key determinant of oxygen-dependent gene regulation. Suppression of HIF-1alpha is important for exploring HIF-1-dependent processes and for interfering with hypoxia-induced pathophysiological events. This study applied RNA-interference targeting HIF-1alpha to the human lung A549 cell line. Transfection of HIF-1alpha-siRNA reduced HIF-1alpha synthesis as measured on mRNA and protein level by realtime RT-PCR, Western blot, and immuncytochemistry. A time kinetic for hypoxic stabilization of HIF-1alpha protein and its inhibition by HIF-1alpha-siRNA is included. Hypoxic induction of HIF-1-controlled target genes as heme oxygenase I (HO-1), phosphoglycerate kinase (PGK), and vascular endothelial growth factor (VEGF) was markedly attenuated by HIF-1alpha-siRNA treatment. Correspondingly, gene activation via hypoxia-responsive-element, as shown by reporter gene assay, was inhibited by HIF-1alpha-siRNA. Moreover, this approach was found to suppress the shift from from S-phase to G1-phase observed in A549 cells in response to hypoxia, supporting a role of HIF-1alpha in oxygen-dependent cell cycle regulation.
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- 2003
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34. Effects of mitochondrial inhibitors and uncouplers on hypoxic vasoconstriction in rabbit lungs.
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Weissmann N, Ebert N, Ahrens M, Ghofrani HA, Schermuly RT, Hänze J, Fink L, Rose F, Conzen J, Seeger W, and Grimminger F
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- 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid pharmacology, Animals, Antifungal Agents pharmacology, Antimycin A pharmacology, Dose-Response Relationship, Drug, In Vitro Techniques, Lung cytology, Methacrylates, Nitric Oxide metabolism, Rabbits, Rotenone pharmacology, Thiazoles pharmacology, omega-N-Methylarginine pharmacology, Enzyme Inhibitors pharmacology, Hypoxia metabolism, Lung physiology, Mitochondria drug effects, Mitochondria metabolism, Uncoupling Agents pharmacology, Vasoconstriction physiology
- Abstract
Hypoxic pulmonary vasoconstriction (HPV) matches lung perfusion to ventilation for optimizing pulmonary gas exchange; however, the underlying mechanism has not yet been fully elucidated. Lung nitric oxide (NO) generation appears to be involved in this process. Recently, mitochondria have been proposed as oxygen sensors, with HPV signaling via a hypoxia-induced increase in the generation of reactive oxygen species derived from mitochondrial complex III and escaping through an anion channel into the cytoplasm. In addition, complex II has been suggested to be specifically involved in hypoxia-dependent generation of reactive oxygen species in the lung. We investigated the effects of several mitochondrial inhibitors and uncouplers on the strength of HPV, and asked for their capacity to mimic HPV during normoxia in isolated buffer-perfused rabbit lungs. Specificity of the agents for HPV was tested by comparison of their effects on non-hypoxia-induced vasoconstriction, elicited by the thromboxane mimetic U46619. Interference with NO metabolism was determined by performing parallel studies with blocked lung NO generation and by measurement of exhaled NO. Rotenone, 3-nitroproprionic acid, and myxothiazol dose-dependently inhibited HPV without being mimics of HPV during normoxia. The inhibitory effect of these agents was only partly specific for HPV by comparison with U46619-induced vasoconstriction. During pre-blocked lung NO synthesis, the selectivity for HPV inhibition was increased for rotenone, but largely lost for myxothiazol. 2-tenoyltrifluoroacetone resulted in an unspecific inhibition of HPV as compared with U46619-induced vasoconstriction. 1-methyl-4-phenylpyridinium iodide and 2-heptyl-4-hydroxyquinoline-N-oxide specifically suppressed HPV and increased normoxic vascular tone. Antimycin A suppressed HPV, an effect being specific in lungs with intact NO synthesis and only partly specific while blocking NO. However, this agent did not mimic HPV during normoxia, as may be expected for interference with the mitochondrial electron transport downstream in complex III. The uncouplers 2,4-dinitrophenol (DNP, 10-200 microM) and carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP, 1-3 microM) induced sustained vasoconstriction during normoxia, with enhancement of HPV by DNP at low and suppression of HPV for both agents at high concentrations. The anion channel blocker 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid inhibited HPV and U46619-induced vasoconstriction with identical dose-response curves. These findings suggest that mitochondria are in some manner involved in the regulation of HPV in intact rabbit lungs. The hypothesis that enhanced superoxide leak at complex III of mitochondria represents the underlying mechanism of acute HPV is supported by the rotenone and 2-heptyl-4-hydroxyquinoline-N-oxide data, but partly contradicted by the findings with 1-methyl-4-phenylpyridinium iodide, antimycin A, DNP, and FCCP. Further studies are mandatory to clarify the link between mitochondrial respiratory chain and hypoxic pulmonary vasoconstriction.
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- 2003
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35. Downregulation of hypoxic vasoconstriction by chronic hypoxia in rabbits: effects of nitric oxide.
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Weissmann N, Nollen M, Gerigk B, Ardeschir Ghofrani H, Schermuly RT, Gunther A, Quanz K, Fink L, Hänze J, Rose F, Seeger W, and Grimminger F
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- 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid pharmacology, Acute Disease, Animals, Breath Tests, Chronic Disease, Disease Models, Animal, Enzyme Inhibitors pharmacology, Female, Hypertension, Pulmonary etiology, Hypertension, Pulmonary pathology, Hypertrophy, Right Ventricular etiology, Hypertrophy, Right Ventricular pathology, Hypoxia complications, Hypoxia drug therapy, In Vitro Techniques, Lung blood supply, Lung pathology, Lung physiopathology, Male, Nitric Oxide administration & dosage, Nitric Oxide metabolism, Nitric Oxide Synthase antagonists & inhibitors, Rabbits, Time, Vasoconstrictor Agents pharmacology, omega-N-Methylarginine pharmacology, Hypertension, Pulmonary physiopathology, Hypoxia physiopathology, Nitric Oxide pharmacology, Vasoconstriction drug effects
- Abstract
Hypoxic pulmonary vasoconstriction (HPV) matches lung perfusion to ventilation for optimizing pulmonary gas exchange. Chronic alveolar hypoxia results in vascular remodeling and pulmonary hypertension. Previous studies have reported conflicting results of the effect of chronic alveolar hypoxia on pulmonary vasoreactivity and the contribution of nitric oxide (NO), which may be related to species and strain differences as well as to the duration of chronic hypoxia. Therefore, we investigated the impact of chronic hypoxia on HPV in rabbits, with a focus on lung NO synthesis. After exposure of the animals to normobaric hypoxia (10% O(2)) for 1 day to 10 wk, vascular reactivity was investigated in ex vivo perfused normoxic ventilated lungs. Chronic hypoxia induced right heart hypertrophy and increased normoxic vascular tone within weeks. The vasoconstrictor response to an acute hypoxic challenge was strongly downregulated within 5 days, whereas the vasoconstrictor response to the thromboxane mimetic U-46619 was maintained. The rapid downregulation of HPV was apparently not linked to changes in the lung vascular NO system, detectable in the exhaled gas and by pharmacological blockage of NO synthesis. Treatment of the animals with long-term inhaled NO reduced right heart hypertrophy and partially maintained the reactivity to acute hypoxia, without any impact on the endogenous NO system being noted. We conclude that chronic hypoxia causes rapid downregulation of acute HPV as a specific event, preceding the development of major pulmonary hypertension and being independent of the lung vascular NO system. Long-term NO inhalation partially maintains the strength of the hypoxic vasoconstrictor response.
- Published
- 2003
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36. Serum levels of vascular endothelial growth factor are elevated in patients with obstructive sleep apnea and severe nighttime hypoxia.
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Schulz R, Hummel C, Heinemann S, Seeger W, and Grimminger F
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- Adaptation, Physiological, Analysis of Variance, Case-Control Studies, DNA-Binding Proteins physiology, Enzyme-Linked Immunosorbent Assay, Erythropoietin physiology, Female, Forced Expiratory Volume, Humans, Hypoxia classification, Hypoxia etiology, Hypoxia-Inducible Factor 1, Hypoxia-Inducible Factor 1, alpha Subunit, Logistic Models, Male, Nuclear Proteins physiology, Obesity complications, Oximetry, Oxygen blood, Polysomnography, Recurrence, Sleep Apnea, Obstructive classification, Sleep Apnea, Obstructive etiology, Up-Regulation, Vascular Endothelial Growth Factor A, Vascular Endothelial Growth Factors, Vital Capacity, Endothelial Growth Factors blood, Hypoxia blood, Lymphokines blood, Severity of Illness Index, Sleep Apnea, Obstructive blood, Transcription Factors
- Abstract
Obstructive sleep apnea (OSA) is associated with increased cardiovascular morbidity and mortality; however, some patients with OSA do not develop cardiovascular disease even in the presence of severe nocturnal oxygen desaturations. Vascular endothelial growth factor (VEGF) is a hypoxia-sensitive glycoprotein stimulating neoangiogenesis. We hypothesized that VEGF production is increased in OSA because of repetitive nocturnal hypoxia. Three different groups were investigated: 10 OSA patients with severe nighttime hypoxia (Group A), 10 OSA patients with moderate hypoxia (Group B), and 10 healthy volunteers (Group C). Serum levels of VEGF were measured by ELISA from peripheral venous blood samples obtained at 7 AM. Group A had significantly (p < 0.01) increased VEGF serum levels when compared with Group B and Group C (mean +/- SEM: 410 +/- 77 pg/ml versus 224 +/- 38 pg/ml and 245 +/- 61 pg/ml). The degree of nocturnal oxygen desaturation in OSA significantly correlated with the VEGF concentrations (r = 0.67, p < 0.01). In conclusion, serum levels of VEGF are elevated in severely hypoxic patients with OSA and are related to the degree of nocturnal oxygen desaturation. This might constitute an adaptive mechanism to counterbalance the emergence of OSA-related cardiovascular disease.
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- 2002
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37. Noncanonical HIPPO/MST Signaling via BUB3 and FOXO Drives Pulmonary Vascular Cell Growth and Survival
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Kudryashova, Tatiana V, Dabral, Swati, Nayakanti, Sreenath, Ray, Arnab, Goncharov, Dmitry A, Avolio, Theodore, Shen, Yuanjun, Rode, Analise, Pena, Andressa, Jiang, Lifeng, Lin, Derek, Baust, Jeffrey, Bachman, Timothy N, Graumann, Johannes, Ruppert, Clemens, Guenther, Andreas, Schmoranzer, Mario, Grobs, Yann, Lemay, Sarah Eve, Tremblay, Eve, Breuils-Bonnet, Sandra, Boucherat, Olivier, Mora, Ana L, DeLisser, Horace, Zhao, Jing, Zhao, Yutong, Bonnet, Sébastien, Seeger, Werner, Pullamsetti, Soni S, and Goncharova, Elena A
- Subjects
Medical Physiology ,Biomedical and Clinical Sciences ,Lung ,Rare Diseases ,2.1 Biological and endogenous factors ,Aetiology ,Cardiovascular ,Animals ,Cell Cycle Proteins ,Cell Proliferation ,Cells ,Cultured ,Chromosomal Proteins ,Non-Histone ,Forkhead Transcription Factors ,Hypertension ,Pulmonary ,Hypoxia ,Mammals ,Mechanistic Target of Rapamycin Complex 1 ,Mice ,Myocytes ,Smooth Muscle ,Poly-ADP-Ribose Binding Proteins ,Proteomics ,Proto-Oncogene Proteins c-akt ,Pulmonary Arterial Hypertension ,Pulmonary Artery ,Vascular Remodeling ,animals ,hypoxia ,mice ,proteomic ,pulmonary artery ,Cardiorespiratory Medicine and Haematology ,Clinical Sciences ,Cardiovascular System & Hematology ,Cardiovascular medicine and haematology ,Clinical sciences - Abstract
RationaleThe MSTs (mammalian Ste20-like kinases) 1/2 are members of the HIPPO pathway that act as growth suppressors in adult proliferative diseases. Pulmonary arterial hypertension (PAH) manifests by increased proliferation and survival of pulmonary vascular cells in small PAs, pulmonary vascular remodeling, and the rise of pulmonary arterial pressure. The role of MST1/2 in PAH is currently unknown.ObjectiveTo investigate the roles and mechanisms of the action of MST1 and MST2 in PAH.Methods and resultsUsing early-passage pulmonary vascular cells from PAH and nondiseased lungs and mice with smooth muscle-specific tamoxifen-inducible Mst1/2 knockdown, we found that, in contrast to canonical antiproliferative/proapoptotic roles, MST1/2 act as proproliferative/prosurvival molecules in human PAH pulmonary arterial vascular smooth muscle cells and pulmonary arterial adventitial fibroblasts and support established pulmonary vascular remodeling and pulmonary hypertension in mice with SU5416/hypoxia-induced pulmonary hypertension. By using unbiased proteomic analysis, gain- and loss-of function approaches, and pharmacological inhibition of MST1/2 kinase activity by XMU-MP-1, we next evaluated mechanisms of regulation and function of MST1/2 in PAH pulmonary vascular cells. We found that, in PAH pulmonary arterial adventitial fibroblasts, the proproliferative function of MST1/2 is caused by IL-6-dependent MST1/2 overexpression, which induces PSMC6-dependent downregulation of forkhead homeobox type O 3 and hyperproliferation. In PAH pulmonary arterial vascular smooth muscle cells, MST1/2 acted via forming a disease-specific interaction with BUB3 and supported ECM (extracellular matrix)- and USP10-dependent BUB3 accumulation, upregulation of Akt-mTORC1, cell proliferation, and survival. Supporting our in vitro observations, smooth muscle-specific Mst1/2 knockdown halted upregulation of Akt-mTORC1 in small muscular PAs of mice with SU5416/hypoxia-induced pulmonary hypertension.ConclusionsTogether, this study describes a novel proproliferative/prosurvival role of MST1/2 in PAH pulmonary vasculature, provides a novel mechanistic link from MST1/2 via BUB3 and forkhead homeobox type O to the abnormal proliferation and survival of pulmonary arterial vascular smooth muscle cells and pulmonary arterial adventitial fibroblasts, remodeling and pulmonary hypertension, and suggests new target pathways for therapeutic intervention.
- Published
- 2022
38. Air travel can be safe and well tolerated in patients with clinically stable pulmonary hypertension
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Thamm, Melanie, Voswinckel, Robert, Tiede, Henning, Lendeckel, Friederike, Grimminger, Friedrich, Seeger, Werner, and Ghofrani, Hossein A.
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- 2011
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39. Hypoxic pulmonary hypertension in mice with constitutively active platelet-derived growth factor receptor-β
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Dahal, Bhola K., Heuchel, Rainer, Pullamsetti, Soni Savai, Wilhelm, Jochen, Ghofrani, Hossein A., Weissmann, Norbert, Seeger, Werner, Grimminger, Friedrich, and Schermuly, Ralph T.
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- 2011
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40. Cytochrome P450 epoxygenase‐derived 5,6‐epoxyeicosatrienoic acid relaxes pulmonary arteries in normoxia but promotes sustained pulmonary vasoconstriction in hypoxia.
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Strielkov, Ievgen, Krause, Nicole C., Knoepp, Fenja, Alebrahimdehkordi, Nasim, Pak, Oleg, Garcia, Claudia, Ghofrani, Hossein A., Schermuly, Ralph T., Seeger, Werner, Grimminger, Friedrich, Sommer, Natascha, and Weissmann, Norbert
- Subjects
CYTOCHROME P-450 ,PULMONARY artery ,VASOCONSTRICTION ,EPOXIDE hydrolase ,HYPOXEMIA ,OXYGENASES - Abstract
Aims: The aim of the study was to investigate the role of cytochrome P450 (CYP) epoxygenase‐derived epoxyeicosatrienoic acids (EETs) in sustained hypoxic pulmonary vasoconstriction (HPV). Methods: Vasomotor responses of isolated mouse intrapulmonary arteries (IPAs) were assessed using wire myography. Key findings were verified by haemodynamic measurements in isolated perfused and ventilated mouse lungs. Results: Pharmacological inhibition of EET synthesis with MS‐PPOH, application of the EET antagonist 14,15‐EEZE or deficiency of CYP2J isoforms suppressed sustained HPV. In contrast, knockdown of EET‐degrading soluble epoxide hydrolase or its inhibition with TPPU augmented sustained HPV almost twofold. All EET regioisomers elicited relaxation in IPAs pre‐contracted with thromboxane mimetic U46619. However, in the presence of KCl‐induced depolarization, 5,6‐EET caused biphasic contraction in IPAs and elevation of pulmonary vascular tone in isolated lungs, whereas other regioisomers had no effect. In patch‐clamp experiments, hypoxia elicited depolarization in pulmonary artery smooth muscle cells (PASMCs), and 5,6‐EET evoked inward whole cell currents in PASMCs depolarized to the hypoxic level, but not at their resting membrane potential. Conclusions: The EET pathway substantially contributes to sustained HPV in mouse pulmonary arteries. 5,6‐EET specifically appears to be involved in HPV, as it is the only EET regioisomer able to elicit not only relaxation, but also sustained contraction in these vessels. 5,6‐EET‐induced pulmonary vasoconstriction is enabled by PASMC depolarization, which occurs in hypoxia. The discovery of the dual role of 5,6‐EET in the regulation of pulmonary vascular tone may provide a basis for the development of novel therapeutic strategies for treatment of HPV‐related diseases. [ABSTRACT FROM AUTHOR]
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- 2020
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41. miR-223-IGF-IR signalling in hypoxia- and load-induced right-ventricular failure: a novel therapeutic approach.
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Lei Shi, Kojonazarov, Baktybek, Elgheznawy, Amro, Popp, Rüdiger, Dahal, Bhola Kumar, Böhm, Mario, Pullamsetti, Soni Savai, Ghofrani, Hossein-Ardeschir, Gödecke, Axel, Jungmann, Andreas, Katus, Hugo A., Müller, Oliver J., Schermuly, Ralph T., Fisslthaler, Beate, Seeger, Werner, and Fleming, Ingrid
- Subjects
INSULIN-like growth factor-binding proteins ,HYPOXEMIA ,HEART failure ,PULMONARY hypertension ,DISEASE progression - Abstract
Aims: Pulmonary hypertension is a progressive disease with poor prognosis, characterized by pathological inward remodelling and loss of patency of the lung vasculature. The right ventricle is co-affected by pulmonary hypertension, which triggers events such as hypoxia and/or increased mechanical load. Initially the right ventricle responds with 'adaptive' hypertrophy, which is often rapidly followed by 'maladaptive' changes leading to right heart decompensation and failure, which is the ultimate cause of death. Methods and results: We report here that miR-223 is expressed in the murine lung and right ventricle at higher levels than in the left ventricle. Moreover, lung and right-ventricular miR-223 levels were markedly down-regulated by hypoxia. Correspondingly, increasing right-ventricular load by pulmonary artery banding, induced right-ventricular ischaemia, and the downregulation of miR-223. Lung and right ventricle miR-223 down-regulation were linked with increased expression of the miR-223 target; insulin-like growth factor-I receptor (IGF-IR) and IGF-I downstream signalling. Similarly, miR-223 was decreased and IGF-IR increased in human pulmonary hypertension. Notably in young mice, miR-223 overexpression, the genetic inactivation or pharmacological inhibition of IGF-IR, all attenuated right-ventricular hypertrophy and improved right heart function under conditions of hypoxia or increased afterload. Conclusion: These findings highlight the early role of pulmonary and right-ventricular miR-223 and the IGF-IR in the right heart failure programme initiated by pulmonary hypoxia and increased mechanical load and may lead to the development of novel therapeutic strategies that target the development of PH and right heart failure. [ABSTRACT FROM AUTHOR]
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- 2016
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42. TIAR and TIA-1 mRNA-Binding Proteins Co-aggregate under Conditions of Rapid Oxygen Decline and Extreme Hypoxia and Suppress the HIF-1α Pathway.
- Author
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Gottschald, Oana R., Malec, Viktor, Krasteva, Gabriela, Hasan, Diya, Kamlah, Florentine, Herold, Susanne, Rose, Frank, Seeger, Werner, and Hänze, Jörg
- Abstract
T-cell intracellular antigen (TIA)-1 and TIA-1-related protein (TIAR) are mRNA-binding proteins that can aggregate within granules under specific stress conditions. In this study, we analyzed TIAR/TIA-1 aggregation under different hypoxic conditions, and studied the effects on the hypoxia-inducible factor (HIF)-1α in different cancer cell lines. Under acute and pronounced hypoxic conditions TIAR/TIA-1 co-aggregated to granules and positive co-staining with eIF3η marker suggested these to represent stress granules. In parallel, HIF-1α expression was blocked in cells displaying TIAR/TIA-1 granules. Silencing of TIAR and TIA-1 caused upregulation of HIF-1α expression, as demonstrated by western blot, immunocytochemistry and HIF-1-dependent reporter gene expression. Additionally, a critical region of the 3′ end of the untranslated HIF-1α mRNA with possible adenosine-uridine-rich elements (AREs) was coupled to the luciferase reporter gene, causing downregulation of expression. Employing this reporter construct, inhibition of TIAR by siRNA attenuated the inhibitory cis-effect of this ARE-sequence. Furthermore, immunohistochemical analysis of A549 cell tumor xenografts revealed a nearly complementary expression of HIF-1α and TIAR reflecting the control of HIF-1α expression by TIAR as revealed in the cell culture studies. In sum, rapid and severe hypoxia caused co-aggregation of TIAR/TIA-1 and these proteins suppressed HIF-1α expression. [ABSTRACT FROM PUBLISHER]
- Published
- 2010
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43. Adenylate Kinase 4—A Key Regulator of Proliferation and Metabolic Shift in Human Pulmonary Arterial Smooth Muscle Cells via Akt and HIF-1α Signaling Pathways.
- Author
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Wujak, Magdalena, Veith, Christine, Wu, Cheng-Yu, Wilke, Tessa, Kanbagli, Zeki Ilker, Novoyatleva, Tatyana, Guenther, Andreas, Seeger, Werner, Grimminger, Friedrich, Sommer, Natascha, Schermuly, Ralph Theo, and Weissmann, Norbert
- Subjects
CELLULAR signal transduction ,VASCULAR remodeling ,PULMONARY arterial hypertension ,SMOOTH muscle ,MUSCLE cells ,GLYCOLYSIS ,HYPOXIA-inducible factor 1 ,PULMONARY circulation - Abstract
Increased proliferation of pulmonary arterial smooth muscle cells (PASMCs) in response to chronic hypoxia contributes to pulmonary vascular remodeling in pulmonary hypertension (PH). PH shares numerous similarities with cancer, including a metabolic shift towards glycolysis. In lung cancer, adenylate kinase 4 (AK4) promotes metabolic reprogramming and metastasis. Against this background, we show that AK4 regulates cell proliferation and energy metabolism of primary human PASMCs. We demonstrate that chronic hypoxia upregulates AK4 in PASMCs in a hypoxia-inducible factor-1α (HIF-1α)-dependent manner. RNA interference of AK4 decreases the viability and proliferation of PASMCs under both normoxia and chronic hypoxia. AK4 silencing in PASMCs augments mitochondrial respiration and reduces glycolytic metabolism. The observed effects are associated with reduced levels of phosphorylated protein kinase B (Akt) as well as HIF-1α, indicating the existence of an AK4-HIF-1α feedforward loop in hypoxic PASMCs. Finally, we show that AK4 levels are elevated in pulmonary vessels from patients with idiopathic pulmonary arterial hypertension (IPAH), and AK4 silencing decreases glycolytic metabolism of IPAH-PASMCs. We conclude that AK4 is a new metabolic regulator in PASMCs interacting with HIF-1α and Akt signaling pathways to drive the pro-proliferative and glycolytic phenotype of PH. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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44. Human RELMβ is a mitogenic factor in lung cells and induced in hypoxia
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Renigunta, Aparna, Hild, Christiane, Rose, Frank, Klepetko, Walter, Grimminger, Friedrich, Seeger, Werner, and Hänze, Jörg
- Subjects
HYPOXEMIA ,CELL lines ,FIBROBLASTS ,LUNG diseases - Abstract
Abstract: RELMβ (resistin-like molecule) represents the most related human homologue of mouse RELMα, also known as hypoxic-induced mitogenic factor (HIMF). In this study, we isolated RELMβ cDNA from human lung tissue and performed regulatory and functional expression studies. RELMβ mRNA was upregulated in hypoxia in human lung A549 cell line as well as primary cultured adventitial fibroblasts and smooth muscle cells (SMC) of pulmonary arteries. Upon transfection of a RELMβ encoding expression plasmid into these cells, we observed significant induction of proliferation particularly in SMC and A549 cells, which could be blocked by phosphatidyl-inositol 3-kinase (PI3K) inhibitors LY294002 and wortmannin. The results suggest that human RELMβ may contribute to hypoxic-induced pulmonary vascular remodeling processes or hypoxia related fibrotic lung disease. [Copyright &y& Elsevier]
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- 2006
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45. Evaluating Systolic and Diastolic Cardiac Function in Rodents Using Microscopic Computed Tomography.
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Kojonazarov, Baktybek, Belenkov, Alexander, Shinomiya, Shohei, Wilchelm, Jochen, Kampschulte, Marian, Mizuno, Shiro, Ghofrani, Hossein Ardeschir, Grimminger, Friedrich, Weissmann, Norbert, Seeger, Werner, and Schermuly, Ralph Theo
- Abstract
Supplemental Digital Content is available in the text. Background: The use of microscopic computed tomography to assess the key functional parameters of systolic emptying or diastolic filling in small animals has not been previously reported. The aim of the study was to test whether microscopic computed tomography can assess the dynamics of both left ventricle and right ventricle (RV) diastolic filling and systolic emptying in an experimental model of pulmonary arterial hypertension Methods and Results: The Wistar-Kyoto rats were injected subcutaneously with the VEGF (vascular endothelial growth factor)-receptor inhibitor SU5416 (20 mg/kg body weight) and were then exposed to chronic hypoxia (10% oxygen) for 21 days (SU5416-hypoxia) followed by normoxia for an additional 2 weeks. Thereafter, multiphase cine cardiac images were acquired using a microscopic computed tomography scanner in conjunction with a blood-pool iodinated contrast agent. Examination of the 3-dimensional images of SU5416-hypoxia rats confirmed the presence of severe pulmonary arterial hypertension. Functional parameters that describe the dynamics of ventricular systolic ejection and diastolic filling were calculated. RV peak ejection rate was significantly decreased (P <0.03) in SU5416-hypoxia rats compared with controls. RV peak filling rate had a significant decrease compared with controls (P <0.03), particularly in the early phase of diastole (P <0.03). This was accompanied by increased time to peak filling rate (P <0.03) and total filling time (P <0.06). Spearman analysis between microscopic computed tomography RV diastolic indices and invasively derived RV end-diastolic pressure indicated excellent correlation. Conclusions: We developed a method that allows rapid and accurate assessment of cardiac functional indices and that paves the way for more extensive preclinical cardiovascular research. [ABSTRACT FROM AUTHOR]
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- 2018
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46. The diverging roles of insulin-like growth factor binding proteins in pulmonary arterial hypertension.
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Schlueter, Beate Christiane, Quanz, Karin, Baldauf, Julia, Petrovic, Aleksandar, Ruppert, Clemens, Guenther, Andreas, Gall, Henning, Tello, Khodr, Grimminger, Friedrich, Ghofrani, Hossein-Ardeschir, Weissmann, Norbert, Seeger, Werner, Schermuly, Ralph Theo, and Weiss, Astrid
- Subjects
- *
INSULIN-like growth factor-binding proteins , *PULMONARY arterial hypertension , *SOMATOMEDIN C , *EPIDERMAL growth factor receptors , *PULMONARY hypertension , *ENDOTHELIN receptors , *BONE morphogenetic protein receptors - Abstract
Pulmonary hypertension (PH) is a progressive, severe and to date not curable disease of the pulmonary vasculature. Alterations of the insulin-like growth factor 1 (IGF-1) system are known to play a role in vascular pathologies and IGF-binding proteins (IGFBPs) are important regulators of the bioavailability and function of IGFs. In this study, we show that circulating plasma levels of IGFBP-1, IGFBP-2 and IGFBP-3 are increased in idiopathic pulmonary arterial hypertension (IPAH) patients compared to healthy individuals. These binding proteins inhibit the IGF-1 induced IGF-1 receptor (IGF1R) phosphorylation and exhibit diverging effects on the IGF-1 induced signaling pathways in human pulmonary arterial cells (i.e. healthy as well as IPAH-hPASMCs, and healthy hPAECs). Furthermore, IGFBPs are differentially expressed in an experimental mouse model of PH. In hypoxic mouse lungs, IGFBP-1 mRNA expression is decreased whereas the mRNA for IGFBP-2 is increased. In contrast to IGFBP-1, IGFBP-2 shows vaso-constrictive properties in the murine pulmonary vasculature. Our analyses show that IGFBP-1 and IGFBP-2 exhibit diverging effects on IGF-1 signaling and display a unique IGF1R-independent kinase activation pattern in human pulmonary arterial smooth muscle cells (hPASMCs), which represent a major contributor of PAH pathobiology. Furthermore, we could show that IGFBP-2, in contrast to IGFBP-1, induces epidermal growth factor receptor (EGFR) signaling, Stat-3 activation and expression of Stat-3 target genes. Based on our results, we conclude that the IGFBP family, especially IGFBP-1, IGFBP-2 and IGFBP-3, are deregulated in PAH, that they affect IGF signaling and thereby regulate the cellular phenotype in PH. [Display omitted] [ABSTRACT FROM AUTHOR]
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- 2024
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47. Hypoxia induces Kv channel current inhibition by increased NADPH oxidase-derived reactive oxygen species
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Mittal, Manish, Gu, Xiang Q., Pak, Oleg, Pamenter, Matthew E., Haag, Daniela, Fuchs, D. Beate, Schermuly, Ralph T., Ghofrani, H.A., Brandes, Ralf P., Seeger, Werner, Grimminger, Friedrich, Haddad, Gabriel G., and Weissmann, Norbert
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- *
HYPOXEMIA , *ACTIVE oxygen in the body , *OXIDASES , *NICOTINAMIDE , *PULMONARY hypertension , *LABORATORY mice , *CYSTEINE - Abstract
Abstract: There is current discussion whether reactive oxygen species are up- or downregulated in the pulmonary circulation during hypoxia, from which sources (i.e., mitochondria or NADPH oxidases) they are derived, and what the downstream targets of ROS are. We recently showed that the NADPH oxidase homolog NOX4 is upregulated in hypoxia-induced pulmonary hypertension in mice and contributes to the vascular remodeling in pulmonary hypertension. We here tested the hypothesis that NOX4 regulates Kv channels via an increased ROS formation after prolonged hypoxia. We showed that (1) NOX4 is upregulated in hypoxia-induced pulmonary hypertension in rats and isolated rat pulmonary arterial smooth muscle cells (PASMC) after 3days of hypoxia, and (2) that NOX4 is a major contributor to increased reactive oxygen species (ROS) after hypoxia. Our data indicate colocalization of Kv1.5 and NOX4 in isolated PASMC. The NADPH oxidase inhibitor and ROS scavenger apocynin as well as NOX4 siRNA reversed the hypoxia-induced decrease in Kv current density whereas the protein levels of the channels remain unaffected by siNOX4 treatment. Determination of cysteine oxidation revealed increased NOX4-mediated Kv1.5 channel oxidation. We conclude that sustained hypoxia decreases Kv channel currents by a direct effect of a NOX4-derived increase in ROS. [Copyright &y& Elsevier]
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- 2012
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48. Redox signaling and reactive oxygen species in hypoxic pulmonary vasoconstriction
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Fuchs, Beate, Sommer, Natascha, Dietrich, Alexander, Schermuly, Ralph Theo, Ghofrani, Hossein Ardeschir, Grimminger, Friedrich, Seeger, Werner, Gudermann, Thomas, and Weissmann, Norbert
- Subjects
- *
OXIDATIVE stress , *PULMONARY blood vessels , *CELLULAR signal transduction , *REACTIVE oxygen species , *HYPOXEMIA , *VASOCONSTRICTION , *MOLECULAR biology , *SMOOTH muscle , *MITOCHONDRIA - Abstract
Abstract: Hypoxic pulmonary vasoconstriction (HPV) is an essential physiological mechanism of the lung that matches blood perfusion with alveolar ventilation to optimize gas exchange. Perturbations of HPV, as may occur in pneumonia or adult respiratory distress syndrome, can cause life-threatening hypoxemia. Despite intensive research for decades, the molecular mechanisms of HPV have not been fully elucidated. Reactive oxygen species (ROS) and changes in the cellular redox state are proposed to link O2 sensing and pulmonary arterial smooth muscle cell contraction underlying HPV. In this regard, mitochondria and NAD(P)H oxidases are discussed as sources of ROS. However, there is controversy whether ROS levels decrease or increase during hypoxia. With this background we summarize the current knowledge on the role of ROS and redox state in HPV. [ABSTRACT FROM AUTHOR]
- Published
- 2010
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49. Genomic Organization and Regulation of a Human 7-Helix Transmembrane Receptor Which Is Expressed in Pulmonary Epithelial Cells and Induced in Hypoxia
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Hänze, Jörg, Groneberg, David A., Rose, Frank, Hanisch, Anika, Dötsch, Jörg, Peiser, Christian, Seeger, Werner, Rascher, Wolfgang, Fischer, Axel, and Grimminger, Friedrich
- Subjects
- *
G proteins , *EPITHELIAL cells , *IN situ hybridization - Abstract
The genetic, regulatory, and tissue-specific characterization of G-protein-coupled receptors is substantial since it contributes to the identification of the natural ligand which may influence basic physiological processes and cell function. Here we explored the genomic structure of a human orphan seven-transmembrane receptor which presents the human homologue of a receptor which has been controversially identified as a rat adrenomedullin receptor subtype. Based on the cDNA sequence a 3.4 kb genomic DNA fragment was isolated. Sequencing of the fragment and comparison studies revealed an intron of 544 bp in the 5′ untranslated region, followed by a second exon encoding the receptor protein of 404 amino acids. The gene is localized on chromosome 12q. The 5′ regulatory region contains several SP1, AP2, and CAAT sites as well as hypoxia responsive elements (HRE) both in the 5′ and 3′ regulatory region. RT-PCR with intron spanning primers demonstrated mRNA signals in various tissues, especially in lung. Characterizing the histological expression pattern in lung sections by nonisotopic in situ hybridization, a strong signal of receptor mRNA was identified in pulmonary epithelial cells of bronchi and alveoli. Analysis of the two human pulmonary epithelial cell lines, H23 and A549, showed significant mRNA induction of this receptor subtype in hypoxia. [Copyright &y& Elsevier]
- Published
- 2002
- Full Text
- View/download PDF
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