103 results on '"Zeldin, D. C."'
Search Results
2. Antigenic stimulation with cytochrome P450 2J expressed in mouse hepatocellular carcinoma cells regulates host anti-tumour immunity
- Author
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Homma, S., Koido, S., Sagawa, Y., Suzuki, H., Komita, H., Nagasaki, E., Takahara, A., Horiguchi-Yamada, J., Tajiri, H., Zeldin, D. C., and Obata, T.
- Published
- 2009
- Full Text
- View/download PDF
3. Cyclooxygenase Polymorphisms and Risk of Cardiovascular Events: The Atherosclerosis Risk in Communities (ARIC) Study
- Author
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Lee, C R, North, K E, Bray, M S, Couper, D J, Heiss, G, and Zeldin, D C
- Published
- 2008
- Full Text
- View/download PDF
4. ANTIHYPERTENSIVE EFFECT OF SOLUBLE EPOXIDE HYDROLASE (sEH) INHIBITION.
- Author
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Yu, Z., Huse, L. M., Engler, M. M., Hammock, B. D., Zeldin, D. C., and Kroetz, D. L.
- Published
- 2000
5. Inherited human group IVA cytosolic phospholipase A2 deficiency abolishes platelet, endothelial, and leucocyte eicosanoid generation
- Author
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Kirkby, N S, Reed, D M, Edin, M L, Rauzi, F, Mataragka, S, Vojnovic, I, Bishop-Bailey, D, Milne, G L, Longhurst, H, Zeldin, D C, Mitchell, J A, and Warner, T D
- Subjects
lipids (amino acids, peptides, and proteins) - Abstract
Eicosanoids are important vascular regulators, but the phospholipase A2 (PLA2) isoforms supporting their production within the cardiovascular system are not fully understood. To address this, we have studied platelets, endothelial cells, and leukocytes from 2 siblings with a homozygous loss-of-function mutation in group IVA cytosolic phospholipase A2 (cPLA2α). Chromatography/mass spectrometry was used to determine levels of a broad range of eicosanoids produced by isolated vascular cells, and in plasma and urine. Eicosanoid release data were paired with studies of cellular function. Absence of cPLA2α almost abolished eicosanoid synthesis in platelets (e.g., thromboxane A2, control 20.5 ± 1.4 ng/ml vs. patient 0.1 ng/ml) and leukocytes [e.g., prostaglandin E2 (PGE2), control 21.9 ± 7.4 ng/ml vs. patient 1.9 ng/ml], and this was associated with impaired platelet activation and enhanced inflammatory responses. cPLA2α-deficient endothelial cells showed reduced, but not absent, formation of prostaglandin I2 (prostacyclin; control 956 ± 422 pg/ml vs. patient 196 pg/ml) and were primed for inflammation. In the urine, prostaglandin metabolites were selectively influenced by cPLA2α deficiency. For example, prostacyclin metabolites were strongly reduced (18.4% of control) in patients lacking cPLA2α, whereas PGE2 metabolites (77.8% of control) were similar to healthy volunteer levels. These studies constitute a definitive account, demonstrating the fundamental role of cPLA2α to eicosanoid formation and cellular responses within the human circulation.
- Published
- 2015
6. CYP2J2 targeting to endothelial cells attenuates adiposity and vascular dysfunction in mice fed a high-fat diet by reprogramming adipocyte phenotype
- Author
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Abraham, N. G., Sodhi, K., Silvis, A. M., Vanella, L., Favero, Gaia, Rezzani, Rita, Lee, C., Zeldin, D. C., and Schwartzman, M. L.
- Published
- 2014
7. Cyclic AMP-induced changes in membrane conductance ofNecturus gallbladder epithelial cells
- Author
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Zeldin, D. C., Corcia, A., and Armstrong, W. M.
- Published
- 1985
- Full Text
- View/download PDF
8. CYP2J2 targeting to endothelial cells attenuates adiposity and vascular dysfunction in mice fed a high fat diet by reprogramming adipocyte phenotype
- Author
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Abraham, N. G., Zeldin, D. C., Sodhi, K., Silcis, A. M., Favero, Gaia, Rezzani, Rita, Rodella, Luigi Fabrizio, Lee, C., Shapiro, J., and Schwartzman, M. L.
- Published
- 2013
9. EET agonist improves cardiac energy metabolism and heart function by regulating fatty acid oxidation and oxidative stress in infarcted myocardium
- Author
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Cao, J., Peterson, S. J., Favero, Gaia, Rezzani, Rita, Rodella, Luigi Fabrizio, Schwartzman, M. L., Zeldin, D. C., Shapiro, J., and Abraham, N. G.
- Published
- 2013
10. Epoxyeicosanoids stimulate multiorgan metastasis and tumor dormancy escape in mice
- Author
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Panigrahy, D, Edin, M L, Lee, C R, Huang, S, Bielenberg, D R, Butterfield, C E, Barnés, C M, Mammoto, A, Mammoto, T, Luria, A, Benny, O, Chaponis, D M, Dudley, A C, Greene, E R, Vergilio, J A, Pietramaggiori, G, Scherer-Pietramaggiori, S S, Short, S M, Seth, M, Lih, F B, Tomer, K B, Yang, J, Schwendener, R A, Hammock, B D, Falck, J R, Manthati, V L, Ingber, D E, Kaipainen, A, D'Amore, P A, Kieran, M W, Zeldin, D C, University of Zurich, and Panigrahy, D
- Subjects
10061 Institute of Molecular Cancer Research ,cardiovascular system ,570 Life sciences ,biology ,lipids (amino acids, peptides, and proteins) ,2700 General Medicine - Abstract
Epoxyeicosatrienoic acids (EETs) are small molecules produced by cytochrome P450 epoxygenases. They are lipid mediators that act as autocrine or paracrine factors to regulate inflammation and vascular tone. As a result, drugs that raise EET levels are in clinical trials for the treatment of hypertension and many other diseases. However, despite their pleiotropic effects on cells, little is known about the role of these epoxyeicosanoids in cancer. Here, using genetic and pharmacological manipulation of endogenous EET levels, we demonstrate that EETs are critical for primary tumor growth and metastasis in a variety of mouse models of cancer. Remarkably, we found that EETs stimulated extensive multiorgan metastasis and escape from tumor dormancy in several tumor models. This systemic metastasis was not caused by excessive primary tumor growth but depended on endothelium-derived EETs at the site of metastasis. Administration of synthetic EETs recapitulated these results, while EET antagonists suppressed tumor growth and metastasis, demonstrating in vivo that pharmacological modulation of EETs can affect cancer growth. Furthermore, inhibitors of soluble epoxide hydrolase (sEH), the enzyme that metabolizes EETs, elevated endogenous EET levels and promoted primary tumor growth and metastasis. Thus, our data indicate a central role for EETs in tumorigenesis, offering a mechanistic link between lipid signaling and cancer and emphasizing the critical importance of considering possible effects of EET-modulating drugs on cancer.
- Published
- 2012
- Full Text
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11. Genetic Variation in Soluble Epoxide Hydrolase (EPHX2) Is Associated With Forearm Vasodilator Responses in Humans
- Author
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Pretorius, M., Zeldin, D. C., Bartlett, J., Brown, N. J., Schuck, R. N., Williams, S. M., Lee, C. R., and Burch, L. H.
- Abstract
Cytochrome P450-derived epoxyeicosatrienoic acids are potent vasodilators in preclinical models and are hydrolyzed by soluble epoxide hydrolase (EPHX2). Associations between the EPHX2 Lys55Arg and Arg287Gln polymorphisms and cardiovascular disease risk have been reported; however, their impact on vascular function in humans has not been investigated. In 265 volunteers (198 white, 67 black American), forearm blood flow was measured by strain-gauge venous occlusion plethysmography at baseline and in response to bradykinin, methacholine and sodium nitroprusside. Forearm vascular resistance was calculated as mean arterial pressure/forearm blood flow. In white Americans, Lys55Arg genotype was associated with vasodilator response to bradykinin, such that forearm blood flow was significantly lower (P=0.043) and forearm vascular resistance was significantly higher (P=0.013) in Arg55 variant allele carriers compared to wild-type individuals. Significant associations were also observed with methacholine and sodium nitroprusside. In contrast, no relationship was observed in black Americans. In black Americans, Arg287Gln genotype was associated with vasodilator response to bradykinin. Although the difference in forearm blood flow did not reach statistical significance (P=0.058), forearm vascular resistance was significantly lower (P=0.037) in Gln287 variant allele carriers compared to wild-type individuals. Significant associations were also observed with methacholine and sodium nitroprusside. In white Americans, Gln287 variant allele carriers did not exhibit significantly higher forearm blood flow (P=0.128) or lower forearm vascular resistance (P=0.080). Genetic variation in EPHX2 is associated with forearm vasodilator responses in a bradykinin receptor- and endothelium-independent manner, suggesting an important role for soluble epoxide hydrolase in the regulation of vascular function in humans.
- Published
- 2011
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12. Modulation by salt intake of the vascular response mediated through adenosine A(2A) receptor: role of CYP epoxygenase and soluble epoxide hydrolase
- Author
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Nayeem, M A, Zeldin, D C, Boegehold, M A, Morisseau, C, Marowsky, A, Ponnoth, D S, Roush, K P, Falck, J R, and University of Zurich
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2737 Physiology (medical) ,570 Life sciences ,biology ,10050 Institute of Pharmacology and Toxicology ,610 Medicine & health ,1314 Physiology - Published
- 2010
13. Modulation of Allergic Airway Inflammation by the Oral Pathogen Porphyromonas gingivalis
- Author
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Barros, S. P., Voltz, J. W., Schwartz, S., Offenbacher, S., Carey, M. A., Morgan, D. L., Card, J. W., Ferguson, C. D., Bradbury, J. A., Zeldin, D. C., Cohen, E. A., Barrow, D. A., Arbes, S. J., and Flake, G. P.
- Subjects
respiratory system ,respiratory tract diseases - Abstract
Accumulating evidence suggests that bacteria associated with periodontal disease may exert systemic immunomodulatory effects. Although the improvement in oral hygiene practices in recent decades correlates with the increased incidence of asthma in developed nations, it is not known whether diseases of the respiratory system might be influenced by the presence of oral pathogens. The present study sought to determine whether subcutaneous infection with the anaerobic oral pathogen Porphyromonas gingivalis exerts a regulatory effect on allergic airway inflammation. BALB/c mice sensitized and subsequently challenged with ovalbumin exhibited airway hyperresponsiveness to methacholine aerosol and increased airway inflammatory cell influx and Th2 cytokine (interleukin-4 [IL-4], IL-5, and IL-13) content relative to those in nonallergic controls. Airway inflammatory cell and cytokine contents were significantly reduced by establishment of a subcutaneous infection with P. gingivalis prior to allergen sensitization, whereas serum levels of ovalbumin-specific IgE and airway responsiveness were not altered. Conversely, subcutaneous infection initiated after allergen sensitization did not alter inflammatory end points but did reduce airway responsiveness in spite of increased serum IgE levels. These data provide the first direct evidence of a regulatory effect of an oral pathogen on allergic airway inflammation and responsiveness. Furthermore, a temporal importance of the establishment of infection relative to allergen sensitization is demonstrated for allergic outcomes.
- Published
- 2010
- Full Text
- View/download PDF
14. EET-Agonist Regulates Human Mesenchymal Stem Cells-Derived Adipocytes Through Activation of HO-1-pAKT Signaling and a decrease in PPARgamma
- Author
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Kim, D. H., Vanella, Luca, Inoue, K, Burgess, A, Gotlinger, K, Manthati, V. L., Koduru, S. R., Zeldin, D. C., Falck, J. R., Schwartzman, M. L., and Abraham, N. G.
- Published
- 2010
15. Human Enteric Microsomal CYP4F Enzymes O-Demethylate the Antiparasitic Prodrug Pafuramidine
- Author
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Hall, J. E., Tidwell, R. R., Zeldin, D. C., Wu, J. Q., Paine, M. F., Wang, M. Z., Bridges, A. S., and Kornbluth, S.
- Abstract
CYP4F enzymes, including CYP4F2 and CYP4F3B, were recently shown to be the major enzymes catalyzing the initial oxidative O-demethylation of the antiparasitic prodrug pafuramidine (DB289) by human liver microsomes. As suggested by a low oral bioavailability, DB289 could undergo first-pass biotransformation in the intestine, as well as in the liver. Using human intestinal microsomes (HIM), we characterized the enteric enzymes that catalyze the initial O-demethylation of DB289 to the intermediate metabolite, M1. M1 formation in HIM was catalyzed by cytochrome P450 (P450) enzymes, as evidenced by potent inhibition by 1-aminoben-zotriazole and the requirement for NADPH. Apparent Km and Vmax values ranged from 0.6 to 2.4 μM and from 0.02 to 0.89 nmol/min/mg protein, respectively (n = 9). Of the P450 chemical inhibitors evaluated, ketoconazole was the most potent, inhibiting M1 formation by 66%. Two inhibitors of P450-mediated arachidonic acid metabolism, HET0016 (N-hydroxy-N′-(4-n-butyl-2-methylphenyl)formamidine) and 17-octadecynoic acid, inhibited M1 formation in a concentration-dependent manner (up to 95%). Immunoinhibition with an antibody raised against CYP4F2 showed concentration-dependent inhibition of M1 formation (up to 92%), whereas antibodies against CYP3A4/5 and CYP2J2 had negligible to modest effects. M1 formation rates correlated strongly with arachidonic acid ω-hydroxylation rates (r2 = 0.94, P < 0.0001, n = 12) in a panel of HIM that lacked detectable CYP4A11 protein expression. Quantitative Western blot analysis revealed appreciable CYP4F expression in these HIM, with a mean (range) of 7 (3–18) pmol/mg protein. We conclude that enteric CYP4F enzymes could play a role in the first-pass biotransformation of DB289 and other xenobiotics.
- Published
- 2007
- Full Text
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16. CYP4F Enzymes Are the Major Enzymes in Human Liver Microsomes That Catalyze the O-Demethylation of the Antiparasitic Prodrug DB289 [2,5-Bis(4-amidinophenyl)furan-bis-O-methylamidoxime]
- Author
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Saulter, J. Y., Bridges, A. S., Parkinson, O. T., Zeldin, D. C., Hall, J. E., Hall, M., Paine, M. F., Wang, M. Z., Cheung, Y.-L., Boykin, D. W., Tidwell, R. R., Usuki, E., Allen, J. L., Parkinson, A., Loewen, G., and Stephens, C. E.
- Abstract
DB289 [2,5-bis(4-amidinophenyl)furan-bis-O-methylamidoxime] is biotransformed to the potent antiparasitic diamidine DB75 [2,5-bis(4-amidinophenyl) furan] by sequential oxidative O-demethylation and reductive N-dehydroxylation reactions. Previous work demonstrated that the N-dehydroxylation reactions are catalyzed by cytochrome b5/NADH-cytochrome b5 reductase. Enzymes responsible for catalyzing the DB289 O-demethylation pathway have not been identified. We report an in vitro metabolism study to characterize enzymes in human liver microsomes (HLMs) that catalyze the initial O-demethylation of DB289 (M1 formation). Potent inhibition by 1-aminobenzotriazole confirmed that M1 formation is catalyzed by P450 enzymes. M1 formation by HLMs was NADPH-dependent, with a Km and Vmax of 0.5 μM and 3.8 nmol/min/mg protein, respectively. Initial screening showed that recombinant CYP1A1, CYP1A2, and CYP1B1 were efficient catalysts of M1 formation. However, none of these three enzymes was responsible for M1 formation by HLMs. Further screening showed that recombinant CYP2J2, CYP4F2, and CYP4F3B could also catalyze M1 formation. An antibody against CYP4F2, which inhibited both CYP4F2 and CYP4F3B, inhibited 91% of M1 formation by HLMs. Two inhibitors of P450-mediated arachidonic acid metabolism, HET0016 (N-hydroxy-N′-(4-n-butyl-2-methylphenyl)formamidine) and 17-octadecynoic acid, effectively inhibited M1 formation by HLMs. Inhibition studies with ebastine and antibodies against CYP2J2 suggested that CYP2J2 was not involved in M1 formation by HLMs. Additionally, ketoconazole preferentially inhibited CYP4F2, but not CYP4F3B, and partially inhibited M1 formation by HLMs. We conclude that CYP4F enzymes (e.g., CYP4F2, CYP4F3B) are the major enzymes responsible for M1 formation by HLMs. These findings indicate that, in human liver, members of the CYP4F subfamily biotransform not only endogenous compounds but also xenobiotics.
- Published
- 2006
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17. NAG-1/GDF15 Transgenic Mouse Has Less White Adipose Tissue and a Reduced Inflammatory Response
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Kim, J. M., primary, Kosak, J. P., additional, Kim, J. K., additional, Kissling, G., additional, Germolec, D. R., additional, Zeldin, D. C., additional, Bradbury, J. A., additional, Baek, S. J., additional, and Eling, T. E., additional
- Published
- 2013
- Full Text
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18. Questionnaire Predictors of Atopy in a US Population Sample: Findings From the National Health and Nutrition Examination Survey, 2005-2006
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Hoppin, J. A., primary, Jaramillo, R., additional, Salo, P., additional, Sandler, D. P., additional, London, S. J., additional, and Zeldin, D. C., additional
- Published
- 2011
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19. Hormonal Influences on Lung Function and Response to Environmental Agents: Lessons from Animal Models of Respiratory Disease
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Card, J. W., primary and Zeldin, D. C., additional
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- 2009
- Full Text
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20. Cyclooxygenase Polymorphisms and Risk of Cardiovascular Events: The Atherosclerosis Risk in Communities (ARIC) Study
- Author
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Lee, C R, primary, North, K E, additional, Bray, M S, additional, Couper, D J, additional, Heiss, G, additional, and Zeldin, D C, additional
- Published
- 2007
- Full Text
- View/download PDF
21. Differential renal gene expression in prehypertensive and hypertensive spontaneously hypertensive rats
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Seubert, J. M., primary, Xu, F., additional, Graves, J. P., additional, Collins, J. B., additional, Sieber, S. O., additional, Paules, R. S., additional, Kroetz, D. L., additional, and Zeldin, D. C., additional
- Published
- 2005
- Full Text
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22. Effects of physical interventions on house dust mite allergen levels in carpet, bed, and upholstery dust in low-income, urban homes.
- Author
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Vojta, P J, primary, Randels, S P, additional, Stout, J, additional, Muilenberg, M, additional, Burge, H A, additional, Lynn, H, additional, Mitchell, H, additional, O'Connor, G T, additional, and Zeldin, D C, additional
- Published
- 2001
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23. NATIONAL SURVEY OF DUST LEAD HAZARDS AND ALLERGENS IN U.S. HOUSING
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Clickner, R P, primary, Friedman, W, additional, Viet, S M, additional, Marker, D, additional, Broene, P, additional, and Zeldin, D C, additional
- Published
- 1998
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24. The rabbit pulmonary cytochrome P450 arachidonic acid metabolic pathway: characterization and significance.
- Author
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Zeldin, D C, primary, Plitman, J D, additional, Kobayashi, J, additional, Miller, R F, additional, Snapper, J R, additional, Falck, J R, additional, Szarek, J L, additional, Philpot, R M, additional, and Capdevila, J H, additional
- Published
- 1995
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25. Overexpression of CYP2J2 provides protection against doxorubicin-induced cardiotoxicity.
- Author
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Zhang, Y., El-Sikhry, H., Chaudhary, K. R., Batchu, S. N., Shayeganpour, A., Jukar, T. O., Bradbury, J. A., Graves, J. P., DeGraff, L. M., Myers, P., D. C. Rouse, J. Foley, A. Nyska, Zeldin, D. C., and Seubert, J. M.
- Subjects
MEDICAL research ,DOXORUBICIN ,MONOOXYGENASES ,CARDIAC imaging ,METALLOENZYMES ,BLOOD circulation disorders ,ANTHROPOMETRY - Abstract
Human cytochrome P-450 (CYP)2J2 is abundant in heart and active in biosynthesis of epoxyeicosatrienoic acids (EEls). Recently, we demonstrated that these eicosanoid products protect myocardium from ischemia-reperfusion injury. The present study utilized trans- genic (Tr) mice with cardiomyocyte-specific overexpression of human CYP2J2 to investigate protection toward toxicity resulting from acute (0, 5, or 15 mg/kg daily for 3 days, followed by 24-h recovery) or chronic (0, 1.5, or 3.0 mg/kg biweekly for 5 wk, followed by 2-wk recovery) doxorubicin (Dox) administration. Acute treatment resulted in marked elevations of serum lactate dehydrogenase and creatine kinase levels that were significantly greater in wild-type (WT) than CYP2J2 Tr mice. Acute treatment also resulted in less activation of stress response enzymes in CYP2J2 Tr mice (catalase 750% vs. 300% of baseline, caspase-3 235% vs. 165% of baseline in WT vs. CYP2J2 Tr mice). Moreover, CYP2J2 Tr hearts exhibited less Dox-induced cardiomyocytes apoptosis (measured by TUNEL) compared with WT hearts. After chronic treatment, comparable decreases in body weight were observed in WT and CYP2J2 Tr mice. However, cardiac function, assessed by measurement of fractional shortening with M-mode transthoracic echocardiography, was significantly higher in CYP2J2 Tr than WT hearts after chronic Dox treatment (WT 37 ± 2%, CYP2J2 Tr 47 ± 1%). WT mice also had larger increases in β-myosin heavy chain and cardiac ankryin repeat protein compared with CYP2J2 Tr mice. CYP2J2 Tr hearts had a significantly higher rate of Dox metabolism than WT hearts (2.2 ± 0.25 vs. 1.6 ± 0.50 ng·min
-1 ·100 μg protein-1 ). In vitro data from H9c2 cells demonstrated that EEls attenuated Dox-induced mitochondrial damage. Together, these data suggest that cardiac-specific overexpression of CYP2J2 limited Dox-induced toxicity. [ABSTRACT FROM AUTHOR]- Published
- 2009
- Full Text
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26. Differential renal gene expression in prehypertensive and hypertensive spontaneously hypertensive rats.
- Author
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Seubert, M., Xu, F., Graves, J. P., Collins, J. B., Sieber, O., Paules, R. S., Kroetz, D. L., and Zeldin, D. C.
- Subjects
HYPERTENSION ,GENE expression ,CARDIOVASCULAR diseases ,MEDICAL genetics ,RATS - Abstract
Development of hypertension stems from both environmental and genetic factors wherein the kidney plays a central role. Spontaneously hypertensive rats (SHR) and the nonhypertensive Wistar-Kyoto (WKY) controls are widely used as a model for studying hypertension. The present study examined the renal gene expression profiles between SHR and WKY at a prehypertensive stage (3 wk of age) and hypertensive stage (9 wk of age). Additionally, age-related changes in gene expression patterns were examined from 3 to 9 wk in both WKY and SHR. Five to six individual kidney samples of the same experimental group were pooled together, and quadruplicate hybridizations were performed using the National Institute of Environmental Health Sciences Rat version 2.0 Chip, which contains ∼6,700 genes. Twenty two genes were found to be differentially expressed between SHR and WKY at 3 wk of age, and 104 genes were differentially expressed at 9 wk of age. Soluble epoxide hydrolase (Ephx2) was found to be significantly upregulated in SHR at both time points and was the predominant outlier. Conversely, elastase 1 (Ela1) was found to be the predominant gene downregulated in SHR at both time points. Analysis of profiles at 3 vs. 9 wk of age identified 508 differentially expressed genes in WKY rats. In contrast, only 211 genes were found to be differentially expressed during this time period in SHR. The altered gene expression patterns observed in the age-related analysis suggested significant differences in the vascular extracellular matrix system between SHR and WKY kidney. Together, our data highlight the complexity of hypertension and the numerous genes involved in and affected by this condition. [ABSTRACT FROM AUTHOR]
- Published
- 2005
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27. Cyclooxygenase-1 and -2 knockout mice demonstrate increased cardiac ischemia/reperfusion injury but are protected by acute preconditioning.
- Author
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Camitta, M G, Gabel, S A, Chulada, P, Bradbury, J A, Langenbach, R, Zeldin, D C, and Murphy, E
- Published
- 2001
28. Cytochrome P450 2C8 ω3-long-chain polyunsaturated fatty acid metabolites increase mouse retinal pathologic neovascularization--brief report.
- Author
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Shao, Zhuo, Fu, Zhong Jie, Stahl, A., Joyal, Julie Anne, Hatton, Colin Paul, Juan, A., Hurst, C., Evans, L., Cui, Z., Pei, D., Gong, Yan, Xu, D., Tian, K., Bogardus, H., Edin, M. L., Lih, F., Sapieha, P., Chen, Jing, Panigrahy, Dipak, Hellstrom, A., Zeldin, D. C., and Smith, Lois Elaine Hodgson
- Subjects
angiogenesis factor ,pathologic neovascularization ,cytochrome P450 CYP2C8 (human) - Abstract
OBJECTIVE: Regulation of angiogenesis is critical for many diseases. Specifically, pathological retinal neovascularization, a major cause of blindness, is suppressed with dietary ω3-long-chain polyunsaturated fatty acids (ω3LCPUFAs) through antiangiogenic metabolites of cyclooxygenase and lipoxygenase. Cytochrome P450 epoxygenases (CYP2C8) also metabolize LCPUFAs, producing bioactive epoxides, which are inactivated by soluble epoxide hydrolase (sEH) to transdihydrodiols. The effect of these enzymes and their metabolites on neovascularization is unknown. APPROACH AND RESULTS: The mouse model of oxygen-induced retinopathy was used to investigate retinal neovascularization. We found that CYP2C (localized in wild-type monocytes/macrophages) is upregulated in oxygen-induced retinopathy, whereas sEH is suppressed, resulting in an increased retinal epoxide:diol ratio. With a ω3LCPUFA-enriched diet, retinal neovascularization increases in Tie2-driven human-CYP2C8-overexpressing mice (Tie2-CYP2C8-Tg), associated with increased plasma 19,20-epoxydocosapentaenoic acid and retinal epoxide:diol ratio. 19,20-Epoxydocosapentaenoic acids and the epoxide:diol ratio are decreased with overexpression of sEH (Tie2-sEH-Tg). Overexpression of CYP2C8 or sEH in mice does not change normal retinal vascular development compared with their wild-type littermate controls. The proangiogenic role in retina of CYP2C8 with both ω3LCPUFA and ω6LCPUFA and antiangiogenic role of sEH in ω3LCPUFA metabolism were corroborated in aortic ring assays. CONCLUSIONS: Our results suggest that CYP2C ω3LCPUFA metabolites promote retinal pathological angiogenesis. CYP2C8 is part of a novel lipid metabolic pathway influencing retinal neovascularization.
- Published
- 2014
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29. Cyclic AMP-induced changes in membrane conductance of Necturus gallbladder epithelial cells.
- Author
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Zeldin, D., Corcia, A., Armstrong, W., Zeldin, D C, and Armstrong, W M
- Abstract
Enhanced cellular cAMP levels have been shown to increase apical membrane Cl- and HCO3- conductances in epithelia. We found that the phosphodiesterase inhibitor 3-isobutyl-1-methyl-xanthine (IBMX) increases cAMP levels in Necturus gallbladder. We used conventional open-tip and double-barreled Cl- -selective microelectrodes to study the effects of IBMX on membrane conductances and intracellular Cl- activities in gallbladders mounted in a divided chamber and bathed with Ringer's solutions at 23 degrees C and pH 7.4. In HCO3- -free media, 0.1 mM IBMX added to the mucosal medium depolarized the apical membrane potential Va, decreased the fractional resistance FR, and significantly reduced intracellular Cl- activity (aCli). Under control conditions, aCli was above the value corresponding to passive distribution across the apical cell membrane. In media containing 25 mM HCO3-, IBMX caused a small transient hyperpolarization of Va followed by a depolarization not significantly different from that observed in HCO3- -free Ringer's. Removal of mucosal Cl-, Na+ or Ca2+ did not affect the IBMX-induced depolarization in Va. The basolateral membrane of Necturus gallbladder is highly K+ permeable. Increasing serosal K+ from 2.5 to 80 mM, depolarized Va. Mucosal IBMX significantly reduced this depolarization. Addition of 10 mM Ba2+, a K+ channel blocker, to the serosal medium depolarized Va and, essentially, blocked the depolarization induced by IBMX. These results indicate that mucosal IBMX increases apical HCO3- conductance and decreases basolateral K+ conductance in gallbladder epithelial cells via a cAMP-dependent mechanism. The latter effect, not previously reported in epithelial tissues, appears to be the major determinant of the IBMX-induced depolarization of Va. [ABSTRACT FROM AUTHOR]
- Published
- 1985
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30. Cytochrome P450 CYP2J9, a new mouse arachidonic acid omega-1 hydroxylase predominantly expressed in brain.
- Author
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Qu, W, Bradbury, J A, Tsao, C C, Maronpot, R, Harry, G J, Parker, C E, Davis, L S, Breyer, M D, Waalkes, M P, Falck, J R, Chen, J, Rosenberg, R L, and Zeldin, D C
- Abstract
A cDNA encoding a new cytochrome P450 was isolated from a mouse brain library. Sequence analysis reveals that this 1,958-base pair cDNA encodes a 57-58-kDa 502-amino acid polypeptide that is 70-91% identical to CYP2J subfamily P450s and is designated CYP2J9. Recombinant CYP2J9 was co-expressed with NADPH-cytochrome P450 oxidoreductase (CYPOR) in Sf9 cells using a baculovirus system. Microsomes of CYP2J9/CYPOR-transfected cells metabolize arachidonic acid to 19-hydroxyeicosatetraenoic acid (HETE) thus CYP2J9 is enzymologically distinct from other P450s. Northern analysis reveals that CYP2J9 transcripts are present at high levels in mouse brain. Mouse brain microsomes biosynthesize 19-HETE. RNA polymerase chain reaction analysis demonstrates that CYP2J9 mRNAs are widely distributed in brain and most abundant in the cerebellum. Immunoblotting using an antibody raised against human CYP2J2 that cross-reacts with CYP2J9 detects a 56-kDa protein band that is expressed in cerebellum and other brain segments and is regulated during postnatal development. In situ hybridization of mouse brain sections with a CYP2J9-specific riboprobe and immunohistochemical staining with the anti-human CYP2J2 IgG reveals abundant CYP2J9 mRNA and protein in cerebellar Purkinje cells. Importantly, 19-HETE inhibits the activity of recombinant P/Q-type Ca(2+) channels that are known to be expressed preferentially in cerebellar Purkinje cells and are involved in triggering neurotransmitter release. Based on these data, we conclude that CYP2J9 is a developmentally regulated P450 that is abundant in brain, localized to cerebellar Purkinje cells, and active in the biosynthesis of 19-HETE, an eicosanoid that inhibits activity of P/Q-type Ca(2+) channels. We postulate that CYP2J9 arachidonic acid products play important functional roles in the brain.
- Published
- 2001
- Full Text
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31. Identification, functional characterization, and regulation of a new cytochrome P450 subfamily, the CYP2Ns.
- Author
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Oleksiak, M F, Wu, S, Parker, C, Karchner, S I, Stegeman, J J, and Zeldin, D C
- Abstract
The screening of liver and heart cDNA libraries from the teleost Fundulus heteroclitus with degenerate oligonucleotide probes to conserved alpha-helical regions in mammalian P450s resulted in the identification of two cDNAs that together represent a novel P450 subfamily, the CYP2Ns. Northern analysis demonstrated that CYP2N1 transcripts are most abundant in liver and intestine, whereas CYP2N2 mRNAs are most abundant in heart and brain. CYP2N1 and CYP2N2 proteins were co-expressed with NADPH-cytochrome P450 oxidoreductase in Sf9 insect cells, and their ability to metabolize arachidonic acid and xenobiotic substrates was examined. Both CYP2N1 and CYP2N2 metabolize arachidonic acid to epoxyeicosatrienoic acids. Epoxidation is highly regio- and enantioselective with preferential formation of (8R,9S)-epoxyeicosatrienoic acid (optical purities are 91 and 90% for CYP2N1 and CYP2N2, respectively) and (11R, 12S)-epoxyeicosatrienoic acid (optical purities are 92 and 70% for CYP2N1 and CYP2N2, respectively). CYP2N1 and CYP2N2 also catalyze the formation of a variety of hydroxyeicosatetraenoic acids. Both P450s have benzphetamine N-demethylase activities but show minimal alkoxyresorufin O-dealkylase activities. To investigate factors affecting CYP2N expression in vivo, CYP2N transcripts were examined following starvation and/or treatment with 12-O-tetradecanoyl phorbol-13-acetate. Intestinal CYP2N1 mRNAs decrease in starved and/or phorbol ester-treated fish, whereas intestinal CYP2N2 transcripts decrease only following phorbol ester treatment. Interestingly, cardiac CYP2N2 expression decreases following phorbol ester treatment but increases following starvation. These results demonstrate that members of this novel P450 subfamily encode early vertebrate forms of arachidonic acid catalysts that are widely expressed and are regulated by environmental factors. Given the wealth of information on the functional role of P450-derived arachidonate metabolites in mammals, we postulate that CYP2N1 and CYP2N2 products have similar biological functions in early vertebrates. The identity of the mammalian orthologue(s) of the CYP2Ns remains unknown.
- Published
- 2000
32. Molecular cloning, enzymatic characterization, developmental expression, and cellular localization of a mouse cytochrome P450 highly expressed in kidney.
- Author
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Ma, J, Qu, W, Scarborough, P E, Tomer, K B, Moomaw, C R, Maronpot, R, Davis, L S, Breyer, M D, and Zeldin, D C
- Abstract
A cDNA encoding a new cytochrome P450 was isolated from a mouse liver library. Sequence analysis reveals that this 1,886-base pair cDNA encodes a 501-amino acid polypeptide that is 69-74% identical to CYP2J subfamily P450s and is designated CYP2J5. Recombinant CYP2J5 was co-expressed with NADPH-cytochrome P450 oxidoreductase in Sf9 cells using a baculovirus system. Microsomal fractions of CYP2J5/NADPH-cytochrome P450 oxidoreductase-transfected cells metabolize arachidonic acid to 14,15-, 11,12-, and 8, 9-epoxyeicosatrienoic acids and 11- and 15-hydroxyeicosatetraenoic acids (catalytic turnover, 4.5 nmol of product/nmol of cytochrome P450/min at 37 degrees C); thus CYP2J5 is enzymologically distinct. Northern analysis reveals that CYP2J5 transcripts are most abundant in mouse kidney and present at lower levels in liver. Immunoblotting using a polyclonal antibody against a CYP2J5-specific peptide detects a protein with the same electrophoretic mobility as recombinant CYP2J5 most abundantly in mouse kidney microsomes. CYP2J5 is regulated during development in a tissue-specific fashion. In the kidney, CYP2J5 is present before birth and reaches maximal levels at 2-4 weeks of age. In the liver, CYP2J5 is absent prenatally and during the early postnatal period, first appears at 1 week, and then remains relatively constant. Immunohistochemical staining of kidney sections with anti-human CYP2J2 IgG reveals that CYP2J protein(s) are present primarily in the proximal tubules and collecting ducts, sites where the epoxyeicosatrienoic acids are known to modulate fluid/electrolyte transport and mediate hormonal action. In situ hybridization confirms abundant CYP2J5 mRNA within tubules of the renal cortex and outer medulla. Epoxyeicosatrienoic acids are endogenous constituents of mouse kidney thus providing direct evidence for the in vivo metabolism of arachidonic acid by the mouse renal epoxygenase(s). Based on these data, we conclude that CYP2J5 is an enzymologically distinct, developmentally regulated, protein that is localized to specific nephron segments and contributes to the oxidation of endogenous renal arachidonic acid pools. In light of the well documented effects of epoxyeicosatrienoic acids in modulating renal tubular transport processes, we postulate that CYP2J5 products play important functional roles in the kidney.
- Published
- 1999
33. CYP2J subfamily P450s in the lung: expression, localization, and potential functional significance.
- Author
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Zeldin, D C, Foley, J, Ma, J, Boyle, J E, Pascual, J M, Moomaw, C R, Tomer, K B, Steenbergen, C, and Wu, S
- Abstract
Cytochrome P450 (P450) monooxygenases catalyze the epoxidation of arachidonic acid to form epoxyeicosatrienoic acids, which modulate bronchial smooth muscle tone and airway transepithelial ion transport. We recently described a new human P450 arachidonic acid epoxygenase (CYP2J2) and the corresponding rat homologue (CYP2J3). Northern analysis of lung RNA using CYP2J cDNA probes demonstrated that CYP2J2 and CYP2J3 mRNAs were expressed in the lung. Immunoblotting of microsomal fractions prepared from human and rat lungs using a polyclonal antibody raised against recombinant human CYP2J2 revealed a single 56-kDa band confirming abundant pulmonary CYP2J2 and CYP2J3 protein expression. Immunohistochemical analysis of formalin-fixed paraffin-embedded human and rat lung sections using the anti-human CYP2J2 IgG and avidin/biotin/peroxidase detection showed that CYP2J proteins were primarily expressed in ciliated epithelial cells lining the airway. Prominent staining was also noted in nonciliated airway epithelial cells, bronchial and pulmonary vascular smooth muscle cells, pulmonary vascular endothelium, and alveolar macrophages, whereas less intense staining was noted in alveolar epithelial cells. Endogenous epoxyeicosatrienoic acids were detected in both human and rat lung using gas chromatography/mass spectrometry, thus providing direct evidence for the in vivo human and rat pulmonary P450 metabolism of arachidonic acid. Based on these data, we conclude that CYP2J2 and CYP2J3 are abundant pulmonary arachidonic acid epoxygenases and that CYP2J products, the epoxyeicosatrienoic acids, are endogenous constituents of human and rat lung. In addition to known effects on airway smooth muscle tone and transepithelial electrolyte transport, the localization of CYP2J proteins to vascular smooth muscle and endothelium suggests that epoxyeicosatrienoic acids may also be involved in the modulation of pulmonary vascular tone.
- Published
- 1996
34. Molecular cloning, expression, and functional significance of a cytochrome P450 highly expressed in rat heart myocytes.
- Author
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Wu, S, Chen, W, Murphy, E, Gabel, S, Tomer, K B, Foley, J, Steenbergen, C, Falck, J R, Moomaw, C R, and Zeldin, D C
- Abstract
A cDNA encoding a P450 monooxygenase was amplified from reverse transcribed rat heart and liver total RNA by polymerase chain reaction using primers based on the 5'- and 3'-end sequences of two rat pseudogenes, CYP2J3P1 and CYP2J3P2. Sequence analysis revealed that this 1,778-base pair cDNA contained an open reading frame and encoded a new 502 amino acid protein designated CYP2J3. Based on the deduced amino acid sequence, CYP2J3 was approximately 70% homologous to both human CYP2J2 and rabbit CYP2J1. Recombinant CYP2J3 protein was co-expressed with NADPH-cytochrome P450 oxidoreductase in Sf9 insect cells using a baculovirus expression system. Microsomal fractions of CYP2J3/NADPH-cytochrome P450 oxidoreductase-transfected cells metabolized arachidonic acid to 14,15-, 11,12-, and 8, 9-epoxyeicosatrienoic acids and 19-hydroxyeicosatetraenoic acid as the principal reaction products (catalytic turnover, 0.2 nmol of product/nmol of cytochrome P450/min at 37 degrees C). Immunoblotting of microsomal fractions prepared from rat tissues using a polyclonal antibody raised against recombinant CYP2J2 that cross-reacted with CYP2J3 but not with other known rat P450s demonstrated abundant expression of CYP2J3 protein in heart and liver. Immunohistochemical staining of formalin-fixed paraffin-embedded rat heart tissue sections using the anti-CYP2J2 IgG and avidin-biotin-peroxidase detection localized expression of CYP2J3 primarily to atrial and ventricular myocytes. In an isolated-perfused rat heart model, 20 min of global ischemia followed by 40 min of reflow resulted in recovery of only 44 +/- 6% of base-line contractile function. The addition of 5 microM 11, 12-epoxyeicosatrienoic acid to the perfusate prior to global ischemia resulted in a significant 1.6-fold improvement in recovery of cardiac contractility (69 +/- 5% of base line, p = 0.01 versus vehicle alone). Importantly, neither 14,15-epoxyeicosatrienoic acid nor 19-hydroxyeicosatetraenoic acid significantly improved functional recovery following global ischemia, demonstrating the specificity of the biological effect for the 11, 12-epoxyeicosatrienoic acid regioisomer. Based on these data, we conclude that (a) CYP2J3 is one of the predominant enzymes responsible for the oxidation of endogenous arachidonic acid pools in rat heart myocytes and (b) 11,12-epoxyeicosatrienoic acid may play an important functional role in the response of the heart to ischemia.
- Published
- 1997
35. Molecular cloning and expression of CYP2J2, a human cytochrome P450 arachidonic acid epoxygenase highly expressed in heart.
- Author
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Wu, S, Moomaw, C R, Tomer, K B, Falck, J R, and Zeldin, D C
- Abstract
A cDNA encoding a human cytochrome P450 arachidonic acid epoxygenase was isolated from a human liver cDNA library. Sequence analysis revealed that this 1,876-base pair cDNA contained an open reading frame and encoded a new 502-amino acid protein designated CYP2J2. Blot hybridization analysis of RNA prepared from human tissues revealed that CYP2J2 was highly expressed in the heart. Recombinant CYP2J2 protein was prepared using the baculovirus expression system and purified to near electrophoretic homogeneity. The enzyme metabolized arachidonic acid predominantly via olefin epoxidation to all four regioisomeric cis-epoxyeicosatrienoic acids (catalytic turnover 65 pmol of product formed/nmol of cytochrome P450/min at 30 degrees C). Epoxidation of arachidonic acid by CYP2J2 at the 14,15-olefin was highly enantioselective for (14R, 15S)-epoxyeicosatrienoic acid (76% optical purity). Immunoblotting of microsomal fractions prepared from human tissues using a polyclonal antibody raised against the recombinant hemoprotein confirmed primary expression of CYP2J2 protein in human heart. The in vivo significance of CYP2J2 was suggested by documenting the presence of epoxyeicosatrienoic acids in the human heart using gas chromatography/mass spectroscopy. Importantly, the chirality of CYP2J2 products matched that of the epoxyeicosatrienoic acid enantiomers present, in vivo, in human heart. We propose that CYP2J2 is one of the enzymes responsible for epoxidation of endogenous arachidonic acid pools in human heart and that epoxyeicosatrienoic acids may, therefore, play important functional roles in cardiac physiology.
- Published
- 1996
36. CYP450-derived oxylipins mediate inflammatory resolution
- Author
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Gilroy, D, Edin, M L, Bystrom, J, Newson, J, Lih, F B, Stables, M L, Zeldin, D C, and Bishop-Bailey, D
- Abstract
Resolution of inflammation has emerged as an active process in immunobiology, with cells of the mononuclear phagocyte system being critical in mediating efferocytosis and wound debridement and bridging the gap between innate and adaptive immunity. Here we investigated the roles of cytochrome P450 (CYP)-derived epoxy-oxylipins in a well-characterized model of sterile resolving peritonitis in the mouse. Epoxy-oxylipins were produced in a biphasic manner during the peaks of acute (4 h) and resolution phases (24–48 h) of the response. The epoxygenase inhibitor SKF525A (epoxI) given at 24 h selectively inhibited arachidonic acid- and linoleic acid-derived CYP450-epoxy-oxlipins and resulted in a dramatic influx in monocytes. The epoxI-recruited monocytes were strongly GR1+, Ly6chi, CCR2hi, CCL2hi, and CX3CR1lo. In addition, expression of F4/80 and the recruitment of T cells, B cells, and dendritic cells were suppressed. sEH (Ephx2)−/− mice, which have elevated epoxy-oxylipins, demonstrated opposing effects to epoxI-treated mice: reduced Ly6chi monocytes and elevated F4/80hi macrophages and B, T, and dendritic cells. Ly6chi and Ly6clo monocytes, resident macrophages, and recruited dendritic cells all showed a dramatic change in their resolution signature following in vivo epoxI treatment. Markers of macrophage differentiation CD11b, MerTK, and CD103 were reduced, and monocyte-derived macrophages and resident macrophages ex vivo showed greatly impaired phagocytosis of zymosan and efferocytosis of apoptotic thymocytes following epoxI treatment. These findings demonstrate that epoxy-oxylipins have a critical role in monocyte lineage recruitment and activity to promote inflammatory resolution and represent a previously unidentified internal regulatory system governing the establishment of adaptive immunity.
37. Basal and inducible anti-inflammatory epoxygenase activity in endothelial cells
- Author
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Askari, A A, Thomson, S, Edin, M L, Lih, F B, Zeldin, D C, and Bishop-Bailey, D
- Abstract
The roles of CYP lipid-metabolizing pathways in endothelial cells are poorly understood. Human endothelial cells expressed CYP2J2 and soluble epoxide hydrolase (sEH) mRNA and protein. The TLR-4 agonist LPS (1 μg/ml; 24 h) induced CYP2J2 but not sEH mRNA and protein. LC–MS/MS analysis of the stable commonly used human endothelial cell line EA.Hy926 showed active epoxygenase and epoxide hydrolase activity: with arachidonic acid (stable epoxide products 5,6-DHET, and 14,15-DHET), linoleic acid (9,10-EPOME and 12,13-EPOME and their stable epoxide hydrolase products 9,10-DHOME and 12,13-DHOME), docosahexaenoic acid (stable epoxide hydrolase product 19,20-DiHDPA) and eicosapentaenoic acid (stable epoxide hydrolase product 17,18-DHET) being formed. Inhibition of epoxygenases using either SKF525A or MS-PPOH induced TNFα release, but did not affect LPS, IL-1β, or phorbol-12-myristate-13-acetate (PMA)-induced TNFα release. In contrast, inhibition of soluble epoxide hydrolase by AUDA or TPPU inhibited basal, LPS, IL-1β and PMA induced TNFα release, and LPS-induced NFκB p65 nuclear translocation. In conclusion, human endothelial cells contain a TLR-4 regulated epoxygenase CYP2J2 and metabolize linoleic acid > eicosapentaenoic acid > arachidonic acid > docosahexaenoic acid to products with anti-inflammatory activity.
38. Functional characterization of cyclooxygenase-2 polymorphisms
- Author
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Fritsche, E., Seung Baek, King, L. M., Zeldin, D. C., Eling, T. E., and Bell, D. A.
39. Selective inhibition of human CYP 2J2 by ebastine analogs
- Author
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Pierre Lafite, Macherey, A. C., Dijols, S., Dansette, P. M., Zeldin, D. C., and Mansuy, D.
40. Lack of formation of Reissner fiber leads to hydrocephalus.
- Author
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Rodríguez, S., Bátiz, L. F., Ortloff, A. R., Vío, K., Muñoz, R.I., DeGraff, L. M., Graves, J. P., Stumpo, D. J., Blackshear, P. J., Zeldin, D. C., Goto, J., Tezuka, T., Yamamoto, T., and Rodríguez, E. M.
- Subjects
HYDROCEPHALUS ,SUBCOMMISSURAL organ ,GLYCOPROTEINS ,CEREBROSPINAL fluid ,IMMUNOGLOBULINS - Abstract
Background The subcommissural organ (SCO) differentiates early in ontogeny and remains fully active during the entire life span. It secretes glycoproteins into the cerebrospinal fluid (CSF) flowing through the Sylvius aqueduct (SA); these proteins either assemble to form Reissner fiber (RF), or remain soluble in the CSF and circulate throughout the CSF compartments. Overholser et al. (1954) have demonstrated that offspring littered by rats maintained on a diet deficient in folic acid and/or Vitamin B12 lack a SCO and develop hydrocephalus. This led them to propose that a dysfunction of the SCO during development leads to stenosis of the SA and hydrocephalus. We have investigated several animal models in which the SCO would play a role in the pathogenesis of hydrocephalus. We have now performed a comparative analysis of the changes occurring in the SCO-RF complex of these animal models, with the aim to find a landmark common to all hydrocephalic animals that might help to unfold the mechanism of SCO-dependant hydrocephalus. Materials and methods A comparative study of the SCO-RF complex and SA of five animal models characterized by a dysfunction of the SCO and development of hydrocephalus was carried out. Conventional light and electron microscopy and immunocytochemistry were applied. Results Model 1. Immunological blockage of RF formation by maternal transfer of antibodies against RF-glycoproteins: undamaged and secretory active SCO, missing RF, stenosed SA and moderate hydrocephalus. Model 2. hyh mice with a point mutation of aSNAP gene: undamaged and secretory active SCO, missing RF, obliterated SA and severe hydrocephalus. Model 3. HTx rat: subcommissural portion (two distal thirds) of SCO missing, supracommissural portion (rostral third) of SCO secretory active, RF absent, obliterated SA and severe hydrocephalus. Model 4. Transgenic mice deficient in transcription factor RFX4- v3: subcommissural portion of SCO missing, supracommissural portion of SCO secretory active, RF absent, patent SA and moderate to severe hydrocephalus. Model 5. fyn knockout mice: subcommissural portion of SCO missing, supracommissural portion of SCO secretory active, RF absent, SA with an abnormal shape and severe hydrocephalus. Conclusion 1. The SCO is formed by two zones: the subcommissural and the supracommissural portions. 2. Differentiation of both zones would be controlled by different genes. 3. The subcommissural portion of the SCO is essential for RF formation. Mutant and transgenic animals lacking this portion, although still have a secretory active flow of CSF throughout SA. 6. Absence of RF could cause SA obliteration or a turbulent CSF flow through SA what, in turn, would lead to hydrocephalus. Supported by Fondecyt 1030265 (Chile), the Intramural Program of the NIH, NIEHS (USA), and MEXT (Japan). [ABSTRACT FROM AUTHOR]
- Published
- 2007
- Full Text
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41. CYP epoxygenase 2J2 prevents cardiac fibrosis by suppression of transmission of pro-inflammation from cardiomyocytes to macrophages.
- Author
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Yang L, Ni L, Duan Q, Wang X, Chen C, Chen S, Chaugai S, Zeldin DC, Tang JR, and Wang DW
- Subjects
- 8,11,14-Eicosatrienoic Acid analogs & derivatives, 8,11,14-Eicosatrienoic Acid genetics, 8,11,14-Eicosatrienoic Acid metabolism, Animals, Cytochrome P-450 CYP2J2, Cytochrome P-450 Enzyme System genetics, Endomyocardial Fibrosis genetics, Endomyocardial Fibrosis pathology, Humans, Inflammation genetics, Inflammation metabolism, Inflammation pathology, Macrophages pathology, Mice, Mice, Transgenic, Myocytes, Cardiac pathology, Transcription Factor RelA genetics, Transcription Factor RelA metabolism, Cytochrome P-450 Enzyme System metabolism, Endomyocardial Fibrosis enzymology, Macrophages enzymology, Myocytes, Cardiac enzymology
- Abstract
Cytochrome P450 epoxygenase (CYP450)-derived epoxyeicosatrienoic acids (EETs) are important regulators of cardiac remodeling; but the underlying mechanism remains unclear. The present study aimed to elucidate how EETs regulated cardiac fibrosis in response to isoprenaline (Iso) or angiotensin (Ang) II. Cardiac-specific human CYP2J2 transgenic mice (Tr) and wild-type (WT) C57BL/6 littermates were infused with Iso- or Ang II. Two weeks after infusion, Tr mice showed more alleviative cardiac fibrosis and inflammation compared with WT mice. In vitro, we found Iso or Ang II induced nuclear transfer of NF-κB p65 and inflammatory cytokines expression in cardiomyocytes. Furthermore, inflammation response emerged in macrophages cultured in cardiomyocytes-conditioned medium. When pretreatment with 14,15-EET in cardiomyocytes, the inflammatory response was markedly suppressed and the transmission of inflammation from cardiomyocytes to macrophages was reduced. In conclusion, CYP2J2 and EETs prevent cardiac fibrosis and cardiac dysfunction by suppressing transmission of pro-inflammation from cardiomyocytes to macrophages in heart, suggesting that elevation of EETs level could be a potential strategy to prevent cardiac fibrosis., (Copyright © 2015 Elsevier Inc. All rights reserved.)
- Published
- 2015
- Full Text
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42. Functional characterization of cyclooxygenase-2 polymorphisms.
- Author
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Fritsche E, Baek SJ, King LM, Zeldin DC, Eling TE, and Bell DA
- Subjects
- Anti-Inflammatory Agents, Non-Steroidal pharmacology, Asian People, Black People, Blotting, Western, Cell Line, Cells, Cultured, Chromatography, High Pressure Liquid, Cloning, Molecular, Cyclooxygenase 2, DNA analysis, DNA genetics, Humans, Isomerism, Membrane Proteins, Models, Molecular, Mutagenesis, Site-Directed, Polymorphism, Genetic, Prostaglandins biosynthesis, White People, Isoenzymes genetics, Prostaglandin-Endoperoxide Synthases genetics
- Abstract
Cyclooxygenases (COX)-1 and -2 are the key enzymes in the conversion of arachidonic acid to prostaglandins. COX-2 appears to play an emerging role in inflammation and carcinogenesis. Nonsteroidal anti-inflammatory drugs (NSAIDs) are used for the treatment of numerous diseases and reduce the risk of developing colorectal cancer. Polymorphisms in the COX-2 gene could alter enzyme expression, function, and/or the response to NSAIDs. Therefore, they could modify individual risks for developing cancer and other diseases or the occurrence of side effects or sensitivity toward selective or nonselective COX inhibitors. We sequenced the COX-2 gene of 72 individuals and identified rare polymorphisms in the promoter and the coding region. A COX-2 molecular model was used to locate the coding region polymorphisms relative to functional sites in the protein, and the COX-2 V511A polymorphism was very near to the active site. This variant protein was expressed, and function was evaluated, but no difference was detected in metabolism of the COX-2 substrates, arachidonic acid, linoleic acid, and 2-arachidonyl glycerol, compared with the wild type. The Km values for arachidonic acid showed no differences between the COX-2 wild type and V511A mutant. Inhibition with selective or nonselective COX inhibitors was essentially the same for the two enzymes. The absence of functionally important polymorphisms in the COX-2 gene may suggest that there has been selective pressure against those single nucleotide polymorphisms because of the critical role of this enzyme in maintenance of homeostasis.
- Published
- 2001
43. Polymorphisms in human CYP2C8 decrease metabolism of the anticancer drug paclitaxel and arachidonic acid.
- Author
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Dai D, Zeldin DC, Blaisdell JA, Chanas B, Coulter SJ, Ghanayem BI, and Goldstein JA
- Subjects
- Alleles, Cell Line, Cytochrome P-450 CYP2C8, Genotype, Humans, Metabolic Clearance Rate, Recombinant Proteins genetics, Recombinant Proteins pharmacokinetics, Sequence Analysis, DNA methods, Antineoplastic Agents, Phytogenic pharmacokinetics, Arachidonic Acid pharmacokinetics, Aryl Hydrocarbon Hydroxylases, Cytochrome P-450 Enzyme System genetics, Paclitaxel pharmacokinetics, Polymorphism, Genetic genetics, Steroid 16-alpha-Hydroxylase, Steroid Hydroxylases genetics
- Abstract
Cytochrome P450 (CYP) 2C8 is the principal enzyme responsible for the metabolism of the anti-cancer drug paclitaxel (Taxol). It is also the predominant P450 responsible for the metabolism of arachidonic acid to biologically active epoxyeicosatrienoic acids (EETs) in human liver and kidney. In this study, we describe two new CYP2C8 alleles containing coding changes: CYP2C8*2 has an Ile269Phe substitution in exon 5 and CYP2C8*3 includes both Arg139Lys and Lys399Arg amino acid substitutions in exons 3 and 8. CYP2C8*2 was found only in African-Americans, while CYP2C8*3 occurred primarily in Caucasians. Neither occurred in Asians. The frequency of the CYP2C8*2 allele was 0.18 in African-Americans, and that of CYP2C8*3 was 0.13 in Caucasians. CYP2C8*1 (wild-type), CYP2C8*2 and CYP2C8*3 cDNAs were expressed in Escherichia coli, and the ability of these enzymes to metabolize both paclitaxel and arachidonic acid was assessed. Recombinant CYP2C8*3 was defective in the metabolism of both substrates. The turnover number of CYP2C8*3 for paclitaxel was 15% of CYP2C8*1. CYP2C8*2 had a two-fold higher Km and two-fold lower intrinsic clearance for paclitaxel than CYP2C8*1. CYP2C8*3 was also markedly defective in the metabolism of arachidonic acid to 11,12- and 14,15-EET (turnover numbers 35-40% that of CYP2C8*1). Thus, CYP2C8*3 is defective in the metabolism of two important CYP2C8 substrates: the anticancer drug paclitaxel and the physiologically important compound arachidonic acid. This polymorphism has important clinical and physiological implications in individuals homozygous for this allele.
- Published
- 2001
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44. Airway inflammation and responsiveness in prostaglandin H synthase-deficient mice exposed to bacterial lipopolysaccharide.
- Author
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Zeldin DC, Wohlford-Lenane C, Chulada P, Bradbury JA, Scarborough PE, Roggli V, Langenbach R, and Schwartz DA
- Subjects
- Administration, Inhalation, Animals, Bronchoalveolar Lavage Fluid, Chemokines metabolism, Cyclooxygenase 1, Cyclooxygenase 2, Cytokines metabolism, Dinoprostone biosynthesis, Disease Models, Animal, Female, Isoenzymes drug effects, Isoenzymes metabolism, Leukotriene B4 biosynthesis, Lipopolysaccharides administration & dosage, Lung pathology, Male, Membrane Proteins, Mice, Mice, Inbred Strains, Mice, Mutant Strains, Pneumonia chemically induced, Prostaglandin-Endoperoxide Synthases drug effects, Prostaglandin-Endoperoxide Synthases metabolism, Proteins metabolism, Up-Regulation, Isoenzymes genetics, Lipopolysaccharides pharmacology, Pneumonia physiopathology, Prostaglandin-Endoperoxide Synthases genetics
- Abstract
Bacterial lipopolysaccharide (LPS) is a risk factor for exacerbation of asthma and causes airway inflammation. The aim of this study was to examine the effects of disruption of prostaglandin (PG) H synthase (PGHS)-1 and PGHS-2 genes on pulmonary responses to inhaled LPS. PGHS-1(-/-), PGHS-2(-/-), and wild-type (WT) mice were exposed to 4 to 6 microg/m(3) LPS via aerosol. Enhanced pause (PenH), a measure of bronchoconstriction, was assessed using a whole-body plethysmograph before and immediately after a 4-h LPS exposure. Bronchoalveolar lavage (BAL) was performed after LPS exposure to assess inflammatory cells, cytokines/chemokines (tumor necrosis factor-alpha, interleukin-6, and macrophage inflammatory protein-2), and PGE(2). The degree of lung inflammation was scored on hematoxylin-and-eosin-stained sections. PGHS-1 and PGHS-2 protein levels were determined by immunoblotting. All mice exhibited increased PenH and methacholine responsiveness after LPS exposure; however, these changes were much more pronounced in PGHS-1(-/-) and PGHS-2(-/-) mice relative to WT mice (P < 0.05). There were no significant differences in inflammation as assessed by BAL fluid (BALF) cells or lung histology between the genotypes despite reduced BALF cytokines/chemokines and PGE(2) in PGHS-1(-/-) and PGHS-2(-/-) mice relative to WT mice (P < 0.05). PGHS-2 was upregulated more in PGHS-1(-/-) mice compared with WT mice after LPS exposure. We conclude that: (1) airway inflammation and hyperresponsiveness are dissociated in PGHS-1(-/-) and PGHS-2(-/-) mice exposed to LPS; (2) the balance of PGHS-1 and PGHS-2 is important in regulating the functional respiratory responses to inhaled LPS; and (3) neither PGHS-1 nor PGHS-2 is important in regulating basal lung function or the inflammatory responses of the lung to inhaled LPS.
- Published
- 2001
- Full Text
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45. Identification and localization of five CYP2Cs in murine extrahepatic tissues and their metabolism of arachidonic acid to regio- and stereoselective products.
- Author
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Tsao CC, Coulter SJ, Chien A, Luo G, Clayton NP, Maronpot R, Goldstein JA, and Zeldin DC
- Subjects
- Animals, Blotting, Western, Female, Hydroxyeicosatetraenoic Acids metabolism, Immunohistochemistry, Isoenzymes metabolism, Male, Mice, Microsomes enzymology, RNA, Messenger biosynthesis, Recombinant Proteins metabolism, Reverse Transcriptase Polymerase Chain Reaction, Stereoisomerism, Arachidonic Acid metabolism, Cytochrome P-450 Enzyme System metabolism
- Abstract
The CYP2C subfamily has been extensively studied in humans with respect to the metabolism of clinically important drugs, and polymorphisms have been identified in these enzymes. In the present study, a murine model was used to determine the possible physiological functions and extrahepatic distribution of CYP2Cs. Using the reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and immununohistochemistry, this report demonstrates that the mouse CYP2Cs are extensively distributed in extrahepatic tissues and localized to heart muscle, lung Clara and ciliated cells, kidney collecting ducts, the X-zone of female adrenals, reproductive organs, white blood cells, and eyes (in the optic nerve, rods, and cones). RT-PCR, subcloning, and sequencing of the products indicate that each CYP2C has a unique tissue distribution. Four cDNA fragments representing potentially new CYP2Cs were identified, each with its own organ-specific pattern of expression. Using a bacterial cDNA expression system, we found that recombinant proteins for each of the five full-length murine CYP2Cs metabolize arachidonic acid to different regio- and stereospecific products, including epoxyeicosatrienoic acids and hydroxyeicosatetraenoic acids. Regio- and stereospecific metabolites of arachidonic acid have been reported to affect important physiological functions such as inflammation, neutrophil activation, ion transport, cellular proliferation, and vascular tone. Our results suggest that the presence of CYP2C enzymes in heart muscle, aorta, kidney, lung, adrenals, eyes, and reproductive organs could regulate important physiological and/or pathological processes in these tissues.
- Published
- 2001
46. Epoxygenase pathways of arachidonic acid metabolism.
- Author
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Zeldin DC
- Subjects
- Animals, Cytochrome P-450 CYP2J2, Humans, Models, Biological, Models, Chemical, Recombinant Proteins metabolism, Arachidonic Acid metabolism, Arachidonic Acids biosynthesis, Cytochrome P-450 Enzyme System metabolism, Oxygenases metabolism
- Published
- 2001
- Full Text
- View/download PDF
47. Overexpression of cytochrome P450 CYP2J2 protects against hypoxia-reoxygenation injury in cultured bovine aortic endothelial cells.
- Author
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Yang B, Graham L, Dikalov S, Mason RP, Falck JR, Liao JK, and Zeldin DC
- Subjects
- Animals, Arachidonic Acid metabolism, Cattle, Cell Hypoxia, Cells, Cultured, Cytochrome P-450 CYP2J2, Cytochrome P-450 Enzyme System genetics, Endothelium, Vascular metabolism, Lipid Peroxidation physiology, Nitric Oxide metabolism, Oxygenases genetics, Receptor, Angiotensin, Type 1, Receptor, Angiotensin, Type 2, Receptors, Angiotensin metabolism, Superoxides metabolism, Transfection, Cytochrome P-450 Enzyme System metabolism, Endothelium, Vascular enzymology, Oxygen metabolism, Oxygenases metabolism, Protective Agents metabolism
- Abstract
CYP2J2 is abundant in human heart and its arachidonic acid metabolites, the epoxyeicosatrienoic acids (EETs), have potent vasodilatory, antiinflammatory and cardioprotective properties. This study was designed to examine the role of CYP2J2 in hypoxia-reoxygenation-induced injury in cultured bovine aortic endothelial cells (BAECs). Early passage BAECs were exposed to 24-h hypoxia followed by 4-h reoxygenation (HR). HR resulted in cell injury, as indicated by significant increases in lactate dehydrogenase (LDH) release and trypan blue stained cells (p < 0.01) and was associated with a decrease in CYP2J2 protein expression. Transfection of BAECs with the CYP2J2 cDNA resulted in increased CYP2J2 expression and arachidonic acid epoxygenase activity, compared with cells transfected with an irrelevant green fluorescent protein (GFP) cDNA. HR induced significant injury in GFP-transfected BAECs, as indicated by increases in LDH release and trypan blue-stained cells (p < 0.01); however, the HR-induced injury was markedly attenuated in CYP2J2-transfected cells (p < 0.01). HR increased cellular 8-iso-prostaglandin F(2alpha) (p < 0.05), and decreased eNOS expression, L-arginine uptake and conversion, and nitrite production (p < 0.01) in GFP-transfected BAECs. CYP2J2 transfection attenuated the HR-induced increase in 8-iso-prostaglandin F(2alpha) (p < 0.05) and decreased the amount of extracellular superoxide detected by cytochrome c reduction under normoxic conditions (p < 0.05) but did not significantly affect HR-induced decreases in eNOS expression, L-arginine uptake and conversion, and nitrite production. Treatment of BAECs with synthetic EETs and/or epoxide hydrolase inhibitors also showed protective effects against HR injury (p < 0.05). These observations suggest: (1) HR results in endothelial injury and decreased CYP2J2 expression; (2) transfection with the CYP2J2 cDNA protects against HR injury; and (3) the cytoprotective effects of CYP2J2 may be mediated, at least in part, by antioxidant effects.
- Published
- 2001
- Full Text
- View/download PDF
48. Activation of Galpha s mediates induction of tissue-type plasminogen activator gene transcription by epoxyeicosatrienoic acids.
- Author
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Node K, Ruan XL, Dai J, Yang SX, Graham L, Zeldin DC, and Liao JK
- Subjects
- Animals, Aorta, Atropine Derivatives, Cattle, Cells, Cultured, Cyclic AMP metabolism, Cytochrome P-450 CYP2J2, GTP-Binding Protein alpha Subunits, Gi-Go metabolism, Gene Expression Regulation, Enzymologic drug effects, Humans, Polymerase Chain Reaction, Proadifen pharmacology, Promoter Regions, Genetic, Saphenous Vein, Transcription, Genetic drug effects, Transfection, 8,11,14-Eicosatrienoic Acid analogs & derivatives, 8,11,14-Eicosatrienoic Acid pharmacology, Cytochrome P-450 Enzyme System metabolism, Endothelium, Vascular enzymology, GTP-Binding Protein alpha Subunits, Gs metabolism, Gene Expression Regulation, Enzymologic physiology, Oxygenases metabolism, Tissue Plasminogen Activator genetics, Transcription, Genetic physiology
- Abstract
The epoxyeicosatrienoic acids (EETs) are products of cytochrome P450 (CYP) epoxygenases that have vasodilatory and anti-inflammatory properties. Here we report that EETs have additional fibrinolytic properties. In vascular endothelial cells, physiological concentrations of EETs, particularly 11,12-EET, or overexpression of the endothelial epoxygenase, CYP2J2, increased tissue plasminogen activator (t-PA) expression by 2.5-fold without affecting plasminogen activator inhibitor-1 expression. This increase in t-PA expression correlated with a 4-fold induction in t-PA gene transcription and a 3-fold increase in t-PA fibrinolytic activity and was blocked by the CYP inhibitor, SKF525A, but not by the calcium-activated potassium channel blocker, charybdotoxin, indicating a mechanism that does not involve endothelial cell hyperpolarization. The t-PA promoter is cAMP-responsive, and induction of t-PA gene transcription by EETs correlated with increases in intracellular cAMP levels and, functionally, with cAMP-driven promoter activity. To determine whether increases in intracellular cAMP levels were due to modulation of guanine nucleotide-binding proteins, we assessed the effects of EETs on Galpha(s) and Galpha(i2). Treatment with EETs increased Galpha(s), but not Galpha(i2), GTP-binding activity by 3.5-fold. These findings indicate that EETs possess fibrinolytic properties through the induction of t-PA and suggest that endothelial CYP2J2 may play an important role in regulating vascular hemostasis.
- Published
- 2001
- Full Text
- View/download PDF
49. The lung HETEs (and EETs) up.
- Author
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Jacobs ER and Zeldin DC
- Subjects
- 8,11,14-Eicosatrienoic Acid metabolism, Animals, Cytochrome P-450 CYP4A, Cytochrome P-450 Enzyme System genetics, Cytochrome P-450 Enzyme System metabolism, Humans, Lung enzymology, Lung ultrastructure, Microsomes enzymology, Microsomes metabolism, Mixed Function Oxygenases genetics, Mixed Function Oxygenases metabolism, Polymorphism, Genetic, Receptors, Cell Surface physiology, 8,11,14-Eicosatrienoic Acid analogs & derivatives, Cytochrome P-450 Enzyme System physiology, Hydroxyeicosatetraenoic Acids physiology, Lung physiology
- Abstract
Arachidonic acid metabolites of the cyclooxygenase and lipoxygenase pathways have a variety of important lung functions. Recent observations indicate that cytochrome P-450 (P-450) monooxygenases are also expressed in the lung, localized to specific pulmonary cell types (e.g., epithelium, endothelium, and smooth muscle), and may modulate critical lung functions. This review summarizes recent data on the presence and biological activity of P-450-derived eicosanoids in the pulmonary vasculature and airways, including effects on pulmonary vascular and bronchial smooth muscle tone and airway epithelial ion transport. We hypothesize a number of potential functions of P-450-derived arachidonate metabolites in the lungs such as contribution to hypoxic pulmonary vasoconstriction, regulation of bronchomotor tone, control of the composition of airway lining fluid, and limitation of pulmonary inflammation. Finally, we describe a number of emerging technologies, including congenic and transgenic strains of experimental animals, P-450 isoform-specific inhibitors and inhibitory antibodies, eicosanoid analogs, and vectors for delivery of P-450 cDNAs and antisense oligonucleotides. These tools will facilitate further studies on the contribution of endogenously formed P-450 eicosanoid metabolites to lung function, under both normal and pathological conditions.
- Published
- 2001
- Full Text
- View/download PDF
50. Soluble epoxide hydrolase regulates hydrolysis of vasoactive epoxyeicosatrienoic acids.
- Author
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Yu Z, Xu F, Huse LM, Morisseau C, Draper AJ, Newman JW, Parker C, Graham L, Engler MM, Hammock BD, Zeldin DC, and Kroetz DL
- Subjects
- 8,11,14-Eicosatrienoic Acid metabolism, 8,11,14-Eicosatrienoic Acid urine, Animals, Arachidonic Acids pharmacology, Blood Pressure drug effects, Cytosol metabolism, Eicosanoids metabolism, Eicosanoids pharmacology, Enzyme Inhibitors pharmacology, Epoxide Hydrolases antagonists & inhibitors, Epoxide Hydrolases pharmacology, Hydrolysis drug effects, Hypertension etiology, Kidney Cortex enzymology, Male, Microsomes enzymology, Microsomes, Liver enzymology, Rats, Rats, Inbred SHR, Rats, Inbred WKY, Rats, Sprague-Dawley, Species Specificity, Urea analogs & derivatives, Urea pharmacology, 8,11,14-Eicosatrienoic Acid analogs & derivatives, Arachidonic Acids metabolism, Epoxide Hydrolases metabolism, Epoxy Compounds metabolism, Hypertension metabolism
- Abstract
The cytochrome P450-derived epoxyeicosatrienoic acids (EETs) have potent effects on renal vascular reactivity and tubular sodium and water transport; however, the role of these eicosanoids in the pathogenesis of hypertension is controversial. The current study examined the hydrolysis of the EETs to the corresponding dihydroxyeicosatrienoic acids (DHETs) as a mechanism for regulation of EET activity and blood pressure. EET hydrolysis was increased 5- to 54-fold in renal cortical S9 fractions from the spontaneously hypertensive rat (SHR) relative to the normotensive Wistar-Kyoto (WKY) rat. This increase was most significant for the 14,15-EET regioisomer, and there was a clear preference for hydrolysis of 14, 15-EET over the 8,9- and 11,12-EETs. Increased EET hydrolysis was consistent with increased expression of soluble epoxide hydrolase (sEH) in the SHR renal microsomes and cytosol relative to the WKY samples. The urinary excretion of 14,15-DHET was 2.6-fold higher in the SHR than in the WKY rat, confirming increased EET hydrolysis in the SHR in vivo. Blood pressure was decreased 22+/-4 mm Hg (P:<0.01) 6 hours after treatment of SHRs with the selective sEH inhibitor N:, N:'-dicyclohexylurea; this treatment had no effect on blood pressure in the WKY rat. These studies identify sEH as a novel therapeutic target for control of blood pressure. The identification of a potent and selective inhibitor of EET hydrolysis will be invaluable in separating the vascular effects of the EET and DHET eicosanoids.
- Published
- 2000
- Full Text
- View/download PDF
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