752 results on '"Dipeptidases"'
Search Results
2. Dysregulation of macrophage PEPD in obesity determines adipose tissue fibro-inflammation and insulin resistance
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V. Pellegrinelli, S. Rodriguez-Cuenca, C. Rouault, E. Figueroa-Juarez, H. Schilbert, S. Virtue, J. M. Moreno-Navarrete, G. Bidault, M. C. Vázquez-Borrego, A. R. Dias, B. Pucker, M. Dale, M. Campbell, S. Carobbio, Y. H. Lin, M. Vacca, J. Aron-Wisnewsky, S. Mora, M. M. Masiero, A. Emmanouilidou, S. Mukhopadhyay, G. Dougan, M. den Hoed, R. J. F. Loos, J. M. Fernández-Real, D. Chiarugi, K. Clément, and A. Vidal-Puig
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medicine.medical_specialty ,Dipeptidases ,Endocrinology, Diabetes and Metabolism ,Adipose tissue ,Inflammation ,Mice ,Insulin resistance ,Internal medicine ,Physiology (medical) ,medicine ,Internal Medicine ,Macrophage ,Animals ,Obesity ,PEPD ,business.industry ,Macrophages ,Cell Biology ,medicine.disease ,Fibrosis ,Endocrinology ,Adipose Tissue ,Diabetes Mellitus, Type 2 ,medicine.symptom ,Insulin Resistance ,business - Abstract
Fibrosis is a hallmark of adipose tissue (AT) dysfunction and obesity-associated insulin resistance that results from an impaired collagen turnover. Peptidase D (PEPD) plays a vital role in collagen turnover by degrading proline-containing dipeptides. Nevertheless, its specific function and importance in AT is unknown. GWAS identified the rs731839 variant in the locus near PEPD that uncouples obesity from insulin resistance and dyslipidaemia, thus indicating that defective PEPD might impair AT remodelling and exacerbate metabolic complications. Here we show that in human and murine obesity, PEPD expression and activity decrease in AT, coupled to the release of PEPD systemically. Both events, in turn, are associated with the accumulation of fibrosis in AT and insulin resistance. Using pharmacologic and genetic animal models of PEPD down-regulation, we show that whereas dysfunctional PEPD activity provokes AT fibrosis, it is the PEPD secreted by AT the main contributor to inflammation, insulin resistance and metabolic dysfunction. Also, PEPD originated in inflammatory macrophages (Mɸ), plays an essential role promoting fibro-inflammatory responses via activation of EGFR in Mɸ and preadipocytes. Using genetic ablation of pepd in Mɸ that prevents obesity-induced PEPD release, also averts AT fibro-inflammation and obesity-associated metabolic dysfunctions. Taking advantage of factor analysis, we have identified the coupling of prolidase decreased activity and increased systemic levels of PEPD as the essential pathogenic triggers of AT fibrosis and insulin resistance. Thus, PEPD produced by Mɸ qualifies as a biomarker of AT fibro-inflammation and a therapeutic target for AT fibrosis and obesity-associated insulin resistance and type 2 diabetes.
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- 2022
3. Enzymatic determination of carnosine in meat and fish using β-Ala-Xaa dipeptidase and histidine ammonia-lyase derived from Pseudomonas putida NBRC100650
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Shinji Nagata, Hisashi Muramatsu, Tomoko Shimamura, Taisuke Harada, Daiki Sugihara, Karen Hashimoto, and Shin-ichiro Kato
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chemistry.chemical_classification ,Dipeptidase ,Dipeptidases ,Meat ,biology ,Pseudomonas putida ,Carnosine ,Fishes ,biology.organism_classification ,Analytical Chemistry ,chemistry.chemical_compound ,Enzyme ,chemistry ,Biochemistry ,biology.protein ,Animals ,%22">Fish ,Cattle ,Histidine Ammonia-Lyase ,Histidine ammonia-lyase - Abstract
Carnosine is a naturally occurring dipeptide and a functional component in foods, while also showing health-promoting effects. Generally, food-derived carnosine is quantified via high-performance liquid chromatography (HPLC). We have developed a method for quantifying carnosine in foods using microbial enzymes, β-Ala-Xaa dipeptidase (BapA) and histidine ammonia-lyase (HAL). The carnosine concentrations in extracts of chicken, pork, beef, bonito, and tuna were determined via both HPLC and enzymatic determination. The carnosine contents measured via enzymatic determination were in agreement with those determined via conventional HPLC analysis. Relative standard-deviation values of the conventional HPLC method and the enzymatic determination of carnosine in foods were 0.728-5.76% and 0.504-4.58%, respectively. The recovery of carnosine in food extracts via enzymatic determination was 97-103%. Therefore, the developed enzymatic determination technique using BapA and HAL can be used for the determination of carnosine in meats and fishes with comparable accuracy to that of conventional HPLC analysis.
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- 2022
4. Carnosine dipeptidase II (CNDP2) protects cells under cysteine insufficiency by hydrolyzing glutathione-related peptides
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Sho Kobayashi, Jia Han, Sohsuke Yamada, Junichi Fujii, Keita Nagaoka, Hideyo Sato, Nobuaki Okumura, Toshifumi Takao, Hiroyuki Konno, and Takujiro Homma
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Dipeptidases ,Kidney ,Carnosine ,Glutathione ,Oxidative phosphorylation ,Fibroblasts ,Biochemistry ,Embryonic stem cell ,Molecular biology ,Mice ,chemistry.chemical_compound ,Cytosol ,medicine.anatomical_structure ,chemistry ,Physiology (medical) ,Extracellular ,medicine ,Animals ,Macrophage ,Cysteine - Abstract
The knockout (KO) of the cystine transporter xCT causes ferroptosis, a type of iron-dependent necrotic cell death, in mouse embryonic fibroblasts, but this does not occur in macrophages. In this study, we explored the gene that supports cell survival under a xCT deficiency using a proteomics approach. Analysis of macrophage-derived peptides that were tagged with iTRAQ by liquid chromatography-mass spectrometry revealed a robust elevation in the levels of carnosine dipeptidase II (CNDP2) in xCT KO macrophages. The elevation in the CNDP2 protein levels was confirmed by immunoblot analyses and this elevation was accompanied by an increase in hydrolytic activity towards cysteinylglycine, the intermediate degradation product of glutathione after the removal of the γ-glutamyl group, in xCT KO macrophages. Supplementation of the cystine-free media of Hepa1-6 cells with glutathione or cysteinylglycine extended their survival, whereas the inclusion of bestatin, an inhibitor of CNDP2, counteracted the effects of these compounds. We established CNDP2 KO mice by means of the CRISPR/Cas9 system and found a decrease in dipeptidase activity in the liver, kidney, and brain. An acetaminophen overdose (350 mg/kg) showed not only aggravated hepatic damage but also renal injury in the CNDP2 KO mice, which was not evident in the wild-type mice that were receiving the same dose. The aggravated renal damage in the CNDP2 KO mice was consistent with the presence of abundant levels of CNDP2 in the kidney, the organ prone to developing ferroptosis. These collective data imply that cytosolic CNDP2, in conjugation with the removal of the γ-glutamyl group, recruits Cys from extracellular GSH and supports redox homeostasis of cells, particularly in epithelial cells of proximal tubules that are continuously exposed to oxidative insult from metabolic wastes that are produced in the body.
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- 2021
5. Crystal structure of aspartyl dipeptidase from <scp> Xenopus laevis </scp> revealed ligand binding induced loop ordering and catalytic triad assembly
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Ashwani Kumar, Biplab Ghosh, Ravindra D. Makde, and R. Singh
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Models, Molecular ,Dipeptidase ,Dipeptidases ,Protein Conformation ,Stereochemistry ,Xenopus ,Crystallography, X-Ray ,Ligands ,Biochemistry ,Xenopus laevis ,Enzyme activator ,chemistry.chemical_compound ,Structural Biology ,Catalytic Domain ,Catalytic triad ,Animals ,Amino Acid Sequence ,Molecular Biology ,chemistry.chemical_classification ,Aspartic Acid ,Binding Sites ,Dipeptide ,biology ,Active site ,Substrate (chemistry) ,biology.organism_classification ,Enzyme ,chemistry ,biology.protein - Abstract
Gene encoding aspartyl dipeptidase from Xenopus levies (PepExl) is upregulated by thyroid hormone and is proposed to play a significant role in resorption of tadpole tail during metamorphosis. However, the importance of peptidase activity for the resorption of the tail remain elusive. Here we report the crystal structures of first eukaryotic S51 peptidase, PepExl, in its ligand-free and Asp-bound states at 1.4 and 1.8 Å resolutions, respectively. The active site is located at dimeric interface and the catalytic triad is found to be dissembled in ligand-free and assembled in Asp-bound state. Structural comparison and molecular dynamic simulations of ligand-free and Asp-bound states shows that distinct loop (loop-A) plays an important role in active site shielding, substrate binding and enzyme activation. This study illuminates the Asp-X dipeptide binding in PepExl is associated with ordering of the loop-A and assembly of residues of catalytic triad in active conformation for enzymatic activity.
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- 2021
6. Influence of carnosine and carnosinase-1 on diabetes-induced afferent arteriole vasodilation: implications for glomerular hemodynamics
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Angelica Rodriguez-Niño, Diego O. Pastene, Steffen A. Hettler, Jiedong Qiu, Thomas Albrecht, Srishti Vajpayee, Rossana Perciaccante, Norbert Gretz, Stephan J. L. Bakker, Bernhard K. Krämer, Benito A. Yard, Jacob van den Born, Groningen Institute for Organ Transplantation (GIOT), and Groningen Kidney Center (GKC)
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RENAL-FUNCTION ,Dipeptidases ,Physiology ,DB/DB MOUSE MODEL ,carnosinase-1 ,Mice, Inbred Strains ,Mice, Transgenic ,HYPERFILTRATION ,MECHANISMS ,Diabetes Mellitus, Experimental ,Mice ,Animals ,Humans ,SINGLE-NEPHRON GFR ,Diabetic Nephropathies ,GLUCOSE-INFUSION ,CLINICAL-SIGNIFICANCE ,Carnosine ,KIDNEY-DISEASE ,Hypertrophy ,diabetic kidney disease ,Vasodilation ,Arterioles ,glomerular hemodynamics ,CNDP1 ,glomerular hyper fi ltration ,LEUCINE REPEAT - Abstract
Dysregulation in glomerular hemodynamics favors hyperfiltration in diabetic kidney disease (DKD). Although carnosine supplementation ameliorates features of DKD, its effect on glomerular vasoregulation is not known. We assessed the influence of carnosine and carnosinase-1 (CN1) on afferent glomerular arteriole vasodilation and its association with glomerular size, hypertrophy, and nephrin expression in diabetic BTBRob/ob mice. Two cohorts of mice including appropriate controls were studied: i.e., diabetic mice that received oral carnosine supplementation (cohort 1) and human (h)CN1 transgenic (TG) diabetic mice (cohort 2). The lumen area ratio (LAR) of the afferent arterioles and glomerular parameters were measured by conventional histology. Three-dimensional analysis using a tissue clearing strategy was also used. In both cohorts, LAR was significantly larger in diabetic BTBRob/ob versus nondiabetic BTBRwt/ob mice (0.41?? 0.05 vs. 0.26 ?? 0.07, P < 0.0001 and 0.42 ?? 0.06 vs. 0.29 ?? 0.04, P < 0.0001) and associated with glomerular size (cohort 1: r = 0.55, P = 0.001 and cohort 2: r = 0.89, P < 0.0001). LAR was partially normalized by oral carnosine supplementation (0.34 ?? 0.05 vs. 0.41?? 0.05, P = 0.004) but did not differ between hCN1 TG and wild-type BTBRob/ob mice. In hCN1 TG mice, serum CN1 concentrations correlated with LAR (r = 0.90, P = 0.006). Diabetic mice displayed decreased nephrin expression and increased glomerular hypertrophy. This was not significantly different in hCN1 TG BTBRob/ob mice (P = 0.06 and P = 0.08, respectively). In conclusion, carnosine and CN1 may affect intraglomerular pressure in an opposing manner through the regulation of afferent arteriolar tone. This study corroborates previous findings on the role of carnosine in the progression of DKD. NEW & NOTEWORTHY Dysregulation in glomerular hemodynamics favors hyperfiltration in diabetic kidney disease (DKD). Although carnosine supplementation ameliorates features of DKD, its effect on glomerular vasoregulation is not known. We assessed the influence of carnosine and carnosinase-1 (CN1) on afferent glomerular arteriole vasodilation and its association with glomerular size, hypertrophy, and nephrin expression in diabetic BTBRob/ob mice. Our results provide evidence that carnosine feeding and CN1 overexpression likely affect intraglomerular pressure through vasoregulation of the afferent arteriole.
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- 2022
7. Dipeptidase-1 governs renal inflammation during ischemia reperfusion injury
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Arthur Lau, Jennifer J. Rahn, Mona Chappellaz, Hyunjae Chung, Hallgrimur Benediktsson, Dominique Bihan, Anne von Mässenhausen, Andreas Linkermann, Craig N. Jenne, Stephen M. Robbins, Donna L. Senger, Ian A. Lewis, Justin Chun, and Daniel A. Muruve
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Inflammation ,Male ,Dipeptidases ,Multidisciplinary ,urogenital system ,Acute Kidney Injury ,GPI-Linked Proteins ,urologic and male genital diseases ,female genital diseases and pregnancy complications ,Mice, Inbred C57BL ,Mice ,Reperfusion Injury ,Animals ,Humans ,Female - Abstract
The mechanisms that drive leukocyte recruitment to the kidney are incompletely understood. Dipeptidase-1 (DPEP1) is a major neutrophil adhesion receptor highly expressed on proximal tubular cells and peritubular capillaries of the kidney. Renal ischemia reperfusion injury (IRI) induces robust neutrophil and monocyte recruitment and causes acute kidney injury (AKI). Renal inflammation and the AKI phenotype were attenuated in Dpep1 −/− mice or mice pretreated with DPEP1 antagonists, including the LSALT peptide, a nonenzymatic DPEP1 inhibitor. DPEP1 deficiency or inhibition primarily blocked neutrophil adhesion to peritubular capillaries and reduced inflammatory monocyte recruitment to the kidney after IRI. CD44 but not ICAM-1 blockade also decreased neutrophil recruitment to the kidney during IRI and was additive to DPEP1 effects. DPEP1, CD44, and ICAM-1 all contributed to the recruitment of monocyte/macrophages to the kidney following IRI. These results identify DPEP1 as a major leukocyte adhesion receptor in the kidney and potential therapeutic target for AKI.
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- 2022
8. Human carnosinase 1 overexpression aggravates diabetes and renal impairment in BTBROb/Ob mice
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Benito A. Yard, Diego O. Pastene, Jiedong Qiu, Stefan Porubsky, Shiqi Zhang, Carolina Delatorre, Bernhard K. Krämer, Harry van Goor, Darya Nosan, Carsten Sticht, Xinmiao Zhang, Thomas Albrecht, Angelica Rodriguez-Niño, Sibylle J. Hauske, Groningen Institute for Organ Transplantation (GIOT), and Groningen Kidney Center (GKC)
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Dipeptidases ,Gene Expression ,Mice, Obese ,Carnosine ,Diabetic nephropathy ,Kidney ,DISEASE ,Antioxidants ,PROTECTS ,Mice ,chemistry.chemical_compound ,0302 clinical medicine ,Drug Discovery ,Transgenic mice ,Diabetic Nephropathies ,Genetics (clinical) ,Proteinuria ,NOCTURNIN ,Immunohistochemistry ,medicine.anatomical_structure ,SECRETION ,Molecular Medicine ,Original Article ,medicine.symptom ,Glycosuria ,medicine.medical_specialty ,CNDP1 GENOTYPE ,NEPHROPATHY ,Mice, Transgenic ,METABOLISM ,Diabetes Mellitus, Experimental ,Nephropathy ,03 medical and health sciences ,Internal medicine ,medicine ,Animals ,Humans ,business.industry ,GLYCOSYLATION ,Gene Expression Profiling ,Computational Biology ,medicine.disease ,GENE ,Renal corpuscle ,Disease Models, Animal ,Endocrinology ,chemistry ,Glycated hemoglobin ,business ,Biomarkers ,030215 immunology - Abstract
Objective To assess the influence of serum carnosinase (CN1) on the course of diabetic kidney disease (DKD). Methods hCN1 transgenic (TG) mice were generated in a BTBROb/Ob genetic background to allow the spontaneous development of DKD in the presence of serum carnosinase. The influence of serum CN1 expression on obesity, hyperglycemia, and renal impairment was assessed. We also studied if aggravation of renal impairment in hCN1 TG BTBROb/Ob mice leads to changes in the renal transcriptome as compared with wild-type BTBROb/Ob mice. Results hCN1 was detected in the serum and urine of mice from two different hCN1 TG lines. The transgene was expressed in the liver but not in the kidney. High CN1 expression was associated with low plasma and renal carnosine concentrations, even after oral carnosine supplementation. Obese hCN1 transgenic BTBROb/Ob mice displayed significantly higher levels of glycated hemoglobin, glycosuria, proteinuria, and increased albumin-creatinine ratios (1104 ± 696 vs 492.1 ± 282.2 μg/mg) accompanied by an increased glomerular tuft area and renal corpuscle size. Gene-expression profiling of renal tissue disclosed hierarchical clustering between BTBROb/Wt, BTBROb/Ob, and hCN1 BTBROb/Ob mice. Along with aggravation of the DKD phenotype, 26 altered genes have been found in obese hCN1 transgenic mice; among them claudin-1, thrombospondin-1, nephronectin, and peroxisome proliferator–activated receptor-alpha have been reported to play essential roles in DKD. Conclusions Our data support a role for serum carnosinase 1 in the progression of DKD. Whether this is mainly attributed to the changes in renal carnosine concentrations warrants further studies. Key messages Increased carnosinase 1 (CN1) is associated with diabetic kidney disease (DKD). BTBROb/Ob mice with human CN1 develop a more aggravated DKD phenotype. Microarray revealed alterations by CN1 which are not altered by hyperglycemia. These genes have been described to play essential roles in DKD. Inhibiting CN1 could be beneficial in DKD.
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- 2020
9. Carnosinase-1 overexpression, but not aerobic exercise training, affects the development of diabetic nephropathy in BTBR ob/ob mice
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Kenneth Vanhove, Jan Stautemas, Maxime Hanssens, Thibaux Van der Stede, Junling He, Hans J. Baelde, Shiqi Zhang, Wim Derave, Shahid P Baba, David Hoetker, Frédéric J. Tessier, Mike Howsam, Inge Everaert, Thomas Albrecht, Kim Bakker, and Benito A. Yard
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0301 basic medicine ,Dipeptidases ,Time Factors ,camosine ,Physiology ,Kidney Glomerulus ,carnosinase-1 ,PROGRESSION ,medicine.disease_cause ,SUPPLEMENTATION ,GLUCOSE ,Diabetic nephropathy ,0302 clinical medicine ,Exercise Physiology ,GLYCEMIC CONTROL ,Hyperlipidemia ,Medicine and Health Sciences ,Diabetic Nephropathies ,OXIDATIVE STRESS ,Kidney ,diabetes ,exercise ,Dipeptides ,ASSOCIATION ,Exercise Therapy ,medicine.anatomical_structure ,Enzyme Induction ,030220 oncology & carcinogenesis ,medicine.symptom ,LEUCINE REPEAT ,TRIGLYCERIDES ,medicine.medical_specialty ,Urology ,Mice, Transgenic ,TYPE-2 ,03 medical and health sciences ,Internal medicine ,Diabetes mellitus ,medicine ,Animals ,Humans ,Aerobic exercise ,Histidine ,Obesity ,Exercise physiology ,Muscle, Skeletal ,business.industry ,diabetic nephropathy ,medicine.disease ,carnosine ,Disease Models, Animal ,RENAL-DISEASE ,030104 developmental biology ,Endocrinology ,Albuminuria ,business ,camosinase-1 ,Oxidative stress - Abstract
doi:10.1152/ajprenal.00329.2019. Manipulation of circulating histidine-containing dipeptides (HCD) has been shown to affect the development of diabetes and early-stage diabetic nephropathy (DN). The aim of the present study was to investigate whether such interventions, which potentially alter levels of circulating HCD, also affect the development of advanced -stage DN. 'Iwo interventions, aerobic exercise training and overexpression of the human carnosinase-1 (hCNI) enzyme, were tested. BTBR ob/ob mice were either subjected to aerobic exercise training (20 wk) or genetically manipulated to overexpress hCNi, and different diabetes- and DNrelated markers were compared with control ob/ob and healthy (wild type) mice. An acute exercise study was performed to elucidate the effect of obesity, acute running, and hCNI overexpression on plasma HCD levels. Chronic aerobic exercise training did not affect the development of diabetes or DN, but hCNI overexpression accelerated hyperlipidemia and aggravated the development of alburninuria, mesangial matrix expansion, and glomerular hypertrophy- of ob/ob mice. In line, plasma, kidney, and muscle 11CD were markedly lower in oh/oh versus wild -type mice, and plasma and kidney HCD in particular were lower in ob/ob hCNI versus ob/ob mice but were unaffected by aerobic exercise. In conclusion, advanced glomerular damage is accelerated in mice overexpressing the hCNI enzyme but not protected by chronic exercise training. Interestingly, we showed, for the first time, that the development of DN is closely linked to renal HCD availability. Further research will have to elucidate whether the stimulation of renal HCD levels can he a therapeutic strategy to reduce the risk for developing DN.
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- 2020
10. DPEP1 expression promotes proliferation and survival of leukaemia cells and correlates with relapse in adults with common B cell acute lymphoblastic leukaemia
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Li-Xin Wu, Xiao-Hui Zhang, Hao Jiang, Jia-Min Zhang, Kai-Yan Liu, Guo-Rui Ruan, Lan-Ping Xu, Ya-Zhen Qin, Qian Jiang, Jing Zhang, Yonghuai Feng, Yu-Hong Chen, Xiao-Jun Huang, Yu Wang, Yan Xu, Yan-Rong Liu, Robert Peter Gale, and Bin Jiang
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Male ,Dipeptidases ,Mice, Nude ,GPI-Linked Proteins ,Clinical correlation ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Animals ,Humans ,Medicine ,Cumulative incidence ,B cell ,Cell Proliferation ,Metalloproteinase ,business.industry ,Hematology ,Precursor Cell Lymphoblastic Leukemia-Lymphoma ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,B-cell acute lymphoblastic leukaemia ,Cancer research ,Female ,Neoplasm Recurrence, Local ,business ,030215 immunology - Abstract
Dehydropeptidase-1 (DPEP1) is a zinc-dependent metalloproteinase abnormally expressed in many cancers. However, its potential role in adults with B cell acute lymphoblastic leukaemia (ALL) is unknown. We found that in adults with common B cell ALL high DPEP1, transcript levels at diagnosis were independently associated with an increased cumulative incidence of relapse (CIR) and worse relapse-free survival (RFS) compared with subjects with low transcript levels. We show an increased proliferation and prosurvival role of DPEP1 in B cell ALL cells via regulation of phosphCREB and p53, which may be the biological basis of the clinical correlation we report. Our data implicate DPEP1 expression in the biology of common B cell ALL in adults. We report clinical correlates and provide a potential biological basis for these correlations. If confirmed, analysing DPEP1 transcript levels at diagnosis could help predict therapy outcomes. Moreover, regulation of DPEP1 expression could be a therapy target in B cell ALL.
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- 2020
11. A single genetic locus controls both expression of DPEP1/CHMP1A and kidney disease development via ferroptosis
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Ziyuan Ma, Xiujie Liang, Katalin Susztak, Yuting Guan, Jacklyn N. Hellwege, Zhen Miao, Benjamin F. Voight, Hongbo Liu, Andreas Linkermann, and Hailong Hu
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Dipeptidases ,Science ,Iron ,Quantitative Trait Loci ,Vesicular Transport Proteins ,General Physics and Astronomy ,Genome-wide association study ,Locus (genetics) ,Haploinsufficiency ,Quantitative trait locus ,Biology ,Kidney ,Article ,General Biochemistry, Genetics and Molecular Biology ,Blood Urea Nitrogen ,Mice ,Folic Acid ,Genetic model ,Pyroptosis ,Genetics ,medicine ,Animals ,Ferroptosis ,Humans ,Genetic Predisposition to Disease ,RNA, Messenger ,Gene ,Cell Proliferation ,Gene Editing ,Regulation of gene expression ,Kidney diseases ,Multidisciplinary ,Functional genomics ,General Chemistry ,DNA Methylation ,Physical Chromosome Mapping ,medicine.disease ,Chromatin ,Gene Expression Regulation ,Genetic Loci ,Organ Specificity ,Necroptosis ,Cisplatin ,Genome-Wide Association Study ,Kidney disease - Abstract
Genome-wide association studies (GWAS) have identified loci for kidney disease, but the causal variants, genes, and pathways remain unknown. Here we identify two kidney disease genes Dipeptidase 1 (DPEP1) and Charged Multivesicular Body Protein 1 A (CHMP1A) via the triangulation of kidney function GWAS, human kidney expression, and methylation quantitative trait loci. Using single-cell chromatin accessibility and genome editing, we fine map the region that controls the expression of both genes. Mouse genetic models demonstrate the causal roles of both genes in kidney disease. Cellular studies indicate that both Dpep1 and Chmp1a are important regulators of a single pathway, ferroptosis and lead to kidney disease development via altering cellular iron trafficking., Identifying causal variants and genes is an essential step in interpreting GWAS loci. Here, the authors investigate a kidney disease GWAS locus with functional genomics data, CRISPR editing and mouse experiments to identify DPEP1 and CHMP1A as putative kidney disease genes via ferroptosis.
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- 2021
12. Quantitation of Methionine Sulfoxide in Milk and Milk-Based Beverages—Minimizing Artificial Oxidation by Anaerobic Enzymatic Hydrolysis
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Michael Hellwig and Martin Kölpin
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0106 biological sciences ,Dipeptidases ,food.ingredient ,Protein oxidation ,01 natural sciences ,Beverages ,Leucyl Aminopeptidase ,chemistry.chemical_compound ,Methionine ,food ,Tandem Mass Spectrometry ,Enzymatic hydrolysis ,Caffeic acid ,Animals ,Anaerobiosis ,Gallic acid ,food.beverage ,Chromatography ,Chemistry ,Evaporated milk ,Methionine sulfoxide ,Hydrolysis ,010401 analytical chemistry ,food and beverages ,General Chemistry ,0104 chemical sciences ,Coffee milk ,Milk ,Biocatalysis ,General Agricultural and Biological Sciences ,Oxidation-Reduction ,010606 plant biology & botany - Abstract
Protein oxidation in milk products may entail flavor changes through reactions at methionine residues. However, little is known about the extent of methionine oxidation in milk and milk products. In the present study, a method for quantitation of methionine, methionine sulfoxide, and methionine sulfone by a stable isotope dilution assay using HILIC-ESI-MS/MS was established. For the quantitation of protein-bound analytes, anaerobic enzymatic hydrolysis was optimized to suppress artificial methionine oxidation. Moreover, the method allowed for monitoring of artificial oxidation by coincubation of the labeled probe [2H8]methionine. The percentage of oxidized methionine was low in UHT milk (up to 1.6%) and evaporated milk (up to 8.8%), but higher in beverages such as cocoa milk drinks (up to 19.0%) and coffee milk drinks (up to 32.8%), resulting in methionine sulfoxide concentrations of up to 6.7 g/kg protein in the latter. These products are important dietary sources of methionine sulfoxide. Model studies revealed that methionine residues can be oxidized strongly in the presence of phenolic compounds such as catechin, caffeic acid, and gallic acid, which are present in cocoa and coffee and may account for the high extent of oxidation in commercial samples.
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- 2019
13. Detection of carnosinase-1 in urine of healthy individuals and patients with type 2 diabetes
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Jana D. Braun, Jiedong Qiu, Sibylle J. Hauske, Xia Li, Christina M. Gant, Bernhard K. Krämer, Stephan J. L. Bakker, Anna Herold, Benito A. Yard, Gozewijn D. Laverman, Shiqi Zhang, Angelica Rodriguez-Niño, Thomas Albrecht, Lifestyle Medicine (LM), Groningen Institute for Organ Transplantation (GIOT), and Groningen Kidney Center (GKC)
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0301 basic medicine ,Glycosuria ,Male ,medicine.medical_specialty ,Dipeptidases ,NEPHROPATHY ,Clinical Biochemistry ,Renal function ,Mice, Transgenic ,Carnosinase-1 ,Type 2 diabetes ,Diabetic nephropathy ,Urine ,Kidney ,Biochemistry ,Gastroenterology ,DISEASE ,Nephropathy ,Excretion ,Cohort Studies ,03 medical and health sciences ,Mice ,Internal medicine ,Diabetes mellitus ,medicine ,Animals ,Humans ,Albuminuria ,Aged ,Aged, 80 and over ,030102 biochemistry & molecular biology ,business.industry ,Organic Chemistry ,Middle Aged ,medicine.disease ,Healthy Volunteers ,030104 developmental biology ,Diabetes Mellitus, Type 2 ,CNDP1 ,Linear Models ,SECRETION ,Female ,medicine.symptom ,business ,ENZYMES ,LEUCINE REPEAT - Abstract
Low serum carnosinase (CN-1) concentrations are associated with low risk for development of diabetic nephropathy (DN) in patients with type 2 diabetes (T2D). Although CN-1 is expressed in the kidney, urinary CN-1 (CNU) excretion and its pathological relevance in patients with T2D have not been investigated to date. The present study therefore assessed the extent of CNU excretion in healthy subjects (n = 243) and in patients with T2D (n = 361) enrolled in the DIAbetes and LifEstyle Cohort Twente-1 (DIALECT-1) in relation to functional renal parameters. CNU was detected in a high proportion of healthy individuals, 180 (74%); median CNU excretion was 0.25 mg/24 h [(IQR 0–0.65 mg/24 h]. In patients with T2D the prevalence and extent of CNU increased in parallel with albuminuria (r = 0.59, p p p n = 241), micro- (n = 80) and macroalbuminuria (n = 40), respectively). Patients with estimated glomerular filtration rate (eGFR) 90 ml/min/1.73 m2) (1.36 vs 0.13 mg/24 h, p R2 = 0.37, p
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- 2019
14. Extracellular Prolidase (PEPD) Induces Anabolic Processes through EGFR, β1-integrin, and IGF-1R Signaling Pathways in an Experimental Model of Wounded Fibroblasts
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Wojciech Miltyk, Jerzy Pałka, Weronika Baszanowska, Magdalena Misiura, and Ilona Oscilowska
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Dipeptidases ,medicine.medical_treatment ,EGFR ,PEPD ,wound healing ,Catalysis ,Article ,Receptor, IGF Type 1 ,lcsh:Chemistry ,Inorganic Chemistry ,Mice ,fibroblasts ,medicine ,Animals ,Humans ,Epidermal growth factor receptor ,Physical and Theoretical Chemistry ,Receptor ,β1-integrin ,lcsh:QH301-705.5 ,Molecular Biology ,Protein kinase B ,Spectroscopy ,PI3K/AKT/mTOR pathway ,Skin ,biology ,integumentary system ,Chemistry ,Growth factor ,Integrin beta1 ,Organic Chemistry ,General Medicine ,Computer Science Applications ,Cell biology ,prolidase ,ErbB Receptors ,lcsh:Biology (General) ,lcsh:QD1-999 ,Gene Expression Regulation ,biology.protein ,IGF-1 ,Signal transduction ,Wound healing ,Signal Transduction - Abstract
The role of prolidase (PEPD) as a ligand of the epidermal growth factor receptor (EGFR) was studied in an experimental model of wound healing in cultured fibroblasts. The cells were treated with PEPD (1&ndash, 100 nM) and analysis of cell viability, proliferation, migration, collagen biosynthesis, PEPD activity, and the expressions of EGFR, insulin-like growth factor 1 (IGF-1), and &beta, 1-integrin receptor including downstream signaling proteins were performed. It has been found that PEPD stimulated proliferation and migration of fibroblasts via activation of the EGFR-downstream PI3K/Akt/mTOR signaling pathway. Simultaneously, PEPD stimulated the expression of &beta, 1-integrin and IGF-1 receptors and proteins downstream to these receptors such as FAK, Grb2, and ERK1/2. Collagen biosynthesis was increased in control and &ldquo, wounded&rdquo, fibroblasts under PEPD treatment. The data suggest that PEPD-induced EGFR signaling may serve as a new attempt to therapy wound healing.
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- 2021
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15. Profibrotic mechanisms of DPP8 and DPP9 highly expressed in the proximal renal tubule epithelial cells
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Li Wang, Ke Li, Jinhua Wang, Yan Li, Yuzhan Zhang, Xiaotao Ma, Xiancheng Li, Wei Dong, Tian Yao, Weihao Zhao, Xuefei Tian, Zhao Chen, and Rongguo Fu
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0301 basic medicine ,Male ,Dipeptidases ,Epithelial-Mesenchymal Transition ,Blotting, Western ,Renal function ,Fluorescent Antibody Technique ,Adamantane ,SMAD ,urologic and male genital diseases ,Real-Time Polymerase Chain Reaction ,Cell Line ,Extracellular matrix ,Kidney Tubules, Proximal ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Gene expression ,Biopsy ,medicine ,Animals ,Humans ,Renal Insufficiency, Chronic ,Dipeptidyl-Peptidases and Tripeptidyl-Peptidases ,Pharmacology ,Kidney ,medicine.diagnostic_test ,business.industry ,Dipeptides ,medicine.disease ,Fibrosis ,Mice, Inbred C57BL ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Case-Control Studies ,Cancer research ,Tubulointerstitial fibrosis ,business ,Kidney disease - Abstract
Background DPP8 and DPP9 have been demonstrated to play important roles in multiple diseases. Evidence for increased gene expression of DPP8 and DPP9 in tubulointerstitium was found to be associated with the decline of kidney function in chronic kidney disease (CKD) patients, which was observed in the Nephroseq human database. To examine the role of DPP8 and DPP9 in the tubulointerstitial injury, we determined the efficacy of DPP8 and DPP9 on epithelial-to-mesenchymal transition (EMT) and tubulointerstitial fibrosis (TIF) as well as the underlying mechanisms. Methods We conducted the immunofluorescence of DPP8 and DPP9 in kidney biopsy specimens of CKD patients, established unilateral ureteral obstruction (UUO) animal model, treated with TC-E5007 (a specific inhibitor of both DPP8 and DPP9) or Saxagliptin (positive control) or saline, and HK-2 cells model. Results We observed the significantly increased expression of DPP8 and DPP9 in the renal proximal tubule epithelial cells of CKD patients compared to the healthy control subjects. DPP8/DPP9 inhibitor TC-E5007 could significantly attenuate the EMT and extracellular matrix (ECM) synthesis in UUO mice, all these effects were mediated via interfering with the TGF-β1/Smad signaling. TC-E5007 treatment also presented reduced renal inflammation and improved renal function in the UUO mice compared to the placebo-treated UUO group. Furthermore, the siRNA for DPP8 and DPP9, and TC-E5007 treatment decreased EMT- and ECM-related proteins in TGF-β1-treated HK-2 cells respectively, which could be reversed significantly by transduction with lentivirus-DPP8 and lentivirus-DPP9. Conclusion These data obtained provide evidence that the DPP8 and DPP9 could be potential therapeutic targets against TIF.
- Published
- 2020
16. Prolidase - A protein with many faces
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Piotr Wilk, Manfred S. Weiss, and Elżbieta Wątor
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0301 basic medicine ,Dipeptidases ,Prolidase deficiency ,030102 biochemistry & molecular biology ,Carcinogenesis ,A protein ,General Medicine ,Biology ,medicine.disease ,Biochemistry ,Neoplasm Proteins ,03 medical and health sciences ,Human health ,030104 developmental biology ,Neoplasms ,medicine ,Animals ,Humans ,Prolidase Deficiency ,Loss function - Abstract
Prolidase is a metal-dependent peptidase specialized in the cleavage of dipeptides containing proline or hydroxyproline on their C-termini. Prolidase homologues are found in all kingdoms of life. The importance of prolidase in human health is underlined by a rare hereditary syndrome referred to as Prolidase Deficiency. A growing number of studies highlight the importance of prolidase in various other human conditions, including cancer. Some recent studies link prolidase's activity-independent regulatory role to tumorigenesis. Furthermore, the enzyme or engineered variants have some applications in biotechnology. In this short review, we aim to highlight different aspects of the protein the importance of which is increasingly recognized over the last years.
- Published
- 2020
17. Current Understanding of the Emerging Role of Prolidase in Cellular Metabolism
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Wojciech Miltyk and Magdalena Misiura
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Dipeptidases ,medicine.medical_treatment ,PEPD ,EGFR ,Review ,Catalysis ,lcsh:Chemistry ,Inorganic Chemistry ,Hydroxyproline ,chemistry.chemical_compound ,medicine ,Animals ,Humans ,Epidermal growth factor receptor ,Physical and Theoretical Chemistry ,Receptor ,lcsh:QH301-705.5 ,Molecular Biology ,Spectroscopy ,Cellular compartment ,Receptors, Interferon ,Prolidase deficiency ,biology ,Chemistry ,Growth factor ,Organic Chemistry ,General Medicine ,medicine.disease ,Computer Science Applications ,Cell biology ,prolidase ,ErbB Receptors ,lcsh:Biology (General) ,lcsh:QD1-999 ,biology.protein ,Tumor Suppressor Protein p53 ,cellular metabolism ,Transforming growth factor ,Signal Transduction - Abstract
Prolidase [EC 3.4.13.9], known as PEPD, cleaves di- and tripeptides containing carboxyl-terminal proline or hydroxyproline. For decades, prolidase has been thoroughly investigated, and several mechanisms regulating its activity are known, including the activation of the β1-integrin receptor, insulin-like growth factor 1 receptor (IGF-1) receptor, and transforming growth factor (TGF)-β1 receptor. This process may result in increased availability of proline in the mitochondrial proline cycle, thus making proline serve as a substrate for the resynthesis of collagen, an intracellular signaling molecule. However, as a ligand, PEPD can bind directly to the epidermal growth factor receptor (EGFR, epidermal growth factor receptor 2 (HER2)) and regulate cellular metabolism. Recent reports have indicated that PEPD protects p53 from uncontrolled p53 subcellular activation and its translocation between cellular compartments. PEPD also participates in the maturation of the interferon α/β receptor by regulating its expression. In addition to the biological effects, prolidase demonstrates clinical significance reflected in the disease known as prolidase deficiency. It is also known that prolidase activity is affected in collagen metabolism disorders, metabolic, and oncological conditions. In this article, we review the latest knowledge about prolidase and highlight its biological function, and thus provide an in-depth understanding of prolidase as a dipeptidase and protein regulating the function of key biomolecules in cellular metabolism.
- Published
- 2020
18. Loss of CNDP causes a shorter lifespan and higher sensitivity to oxidative stress in Drosophila melanogaster
- Author
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Kimiko, Yamakawa-Kobayashi, Yuya, Ohhara, Takumi, Kawashima, Yoshitatsu, Ohishi, and Yasunari, Kayashima
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Gene Editing ,Male ,Paraquat ,Dipeptidases ,Base Sequence ,Carnosine ,Longevity ,Gene Expression ,Hydrogen Peroxide ,Glutathione ,Antioxidants ,Animals, Genetically Modified ,Oxidative Stress ,Drosophila melanogaster ,Mutation ,Animals ,Drosophila Proteins ,CRISPR-Cas Systems ,Reactive Oxygen Species - Abstract
Increasing oxidative stress seems to be the result of an imbalance between free radical production and antioxidant defenses. During the course of aging, oxidative stress causes tissue/cellular damage, which is implicated in numerous age-related diseases. Carnosinase (CN or CNDP) is dipeptidase, which is associated with carnosine and/or glutathione (GSH) metabolism, those are the most abundant naturally occurring endogenous dipeptide and tripeptides with antioxidant and free radical scavenger properties. In the present study, we generated Drosophila cndp (dcndp) mutant flies using the CRISPR/Cas9 system to study the roles of dcndp in vivo. We demonstrate that dcndp mutant flies exhibit shorter lifespan and increased sensitivity to paraquat or hydrogen peroxide (H
- Published
- 2020
19. DPP8/9 inhibitors activate the CARD8 inflammasome in resting lymphocytes
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Daniel A. Bachovchin, Elizabeth L. Orth, Marian C. Okondo, Daniel P. Ball, Sahana D. Rao, Darren C. Johnson, and Hsin-Che Huang
- Subjects
Cancer Research ,Programmed cell death ,Cell type ,Proteases ,Dipeptidases ,Inflammasomes ,Immunology ,NLR Proteins ,Lymphocyte Activation ,Article ,Cellular and Molecular Neuroscience ,Mice ,Cell death and immune response ,medicine ,Pyroptosis ,Animals ,Humans ,Protease Inhibitors ,Lymphocytes ,lcsh:QH573-671 ,Dipeptidyl-Peptidases and Tripeptidyl-Peptidases ,Cells, Cultured ,Adaptor Proteins, Signal Transducing ,Innate immune system ,lcsh:Cytology ,Chemistry ,Immune cell death ,Cell Cycle ,Inflammasome ,Cell Biology ,Cell biology ,Neoplasm Proteins ,Rats ,CARD Signaling Adaptor Proteins ,Apoptosis Regulatory Proteins ,CD8 ,Intracellular ,medicine.drug - Abstract
Canonical inflammasomes are innate immune signaling platforms that are formed in response to intracellular pathogen-associated signals and trigger caspase-1-dependent pyroptosis. Inflammasome formation and signaling is thought to mainly occur in myeloid cells, and in particular monocytes and macrophages. Here we show that small molecule inhibitors of dipeptidyl peptidases 8 and 9 (DPP8/9), which activate the related CARD8 and NLRP1 inflammasomes, also activate pyroptosis in human and rodent resting lymphocytes. We found that both CD4+ and CD8+ T cells were particularly sensitive to these inhibitors, although the sensitivity of T cells, like macrophages, varied considerably between species. In human T cells, we show that CARD8 mediates DPP8/9 inhibitor-induced pyroptosis. Intriguingly, although activated human T cells express the key proteins known to be required for CARD8-mediated pyroptosis, these cells were completely resistant to DPP8/9 inhibitors. Overall, these data show that resting lymphoid cells can activate at least one inflammasome, revealing additional cell types and states poised to undergo rapid pyroptotic cell death in response to danger-associated signals.
- Published
- 2020
20. High-throughput mediation analysis of human proteome and metabolome identifies mediators of post-bariatric surgical diabetes control
- Author
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Barbara B. Kahn, Pratik Aryal, Hui Pan, Yixing Yuchi, Yinong Sebastian, Simon Kasif, Donald C. Simonson, Allison B. Goldfine, Cristina M. Rondinone, Mary-Elizabeth Patti, Vera Djordjilović, Anish Konkar, Jonathan M. Dreyfuss, Brandon W. Higgs, Kathleen Foster, Florencia Halperin, Randy J. Seeley, Ashley H. Vernon, Joseph Grimsby, Xuehong Dong, and Vissarion Efthymiou
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Blood Glucose ,Dipeptidases ,Proteome ,Statistical methods ,General Physics and Astronomy ,Type 2 diabetes ,Growth hormone receptor ,Body Mass Index ,Human proteome project ,IGFBP1 ,Gene knockdown ,Multidisciplinary ,Human Growth Hormone ,digestive, oral, and skin physiology ,Fasting ,Liver ,Metabolome ,Settore SECS-S/01 - Statistica ,hormones, hormone substitutes, and hormone antagonists ,Mediation (statistics) ,medicine.medical_specialty ,Science ,Primary Cell Culture ,Gastric Bypass ,Settore BIO/11 - Biologia Molecolare ,General Biochemistry, Genetics and Molecular Biology ,Article ,Diabetes mellitus ,Internal medicine ,medicine ,Animals ,Humans ,Obesity ,Retrospective Studies ,Glycated Hemoglobin ,business.industry ,nutritional and metabolic diseases ,General Chemistry ,medicine.disease ,Rats ,Insulin-Like Growth Factor Binding Protein 1 ,Insulin-Like Growth Factor Binding Protein 2 ,Endocrinology ,Diabetes Mellitus, Type 2 ,Gene Expression Regulation ,Hepatocytes ,business ,Carrier Proteins ,Biomarkers - Abstract
To improve the power of mediation in high-throughput studies, here we introduce High-throughput mediation analysis (Hitman), which accounts for direction of mediation and applies empirical Bayesian linear modeling. We apply Hitman in a retrospective, exploratory analysis of the SLIMM-T2D clinical trial in which participants with type 2 diabetes were randomized to Roux-en-Y gastric bypass (RYGB) or nonsurgical diabetes/weight management, and fasting plasma proteome and metabolome were assayed up to 3 years. RYGB caused greater improvement in HbA1c, which was mediated by growth hormone receptor (GHR). GHR’s mediation is more significant than clinical mediators, including BMI. GHR decreases at 3 months postoperatively alongside increased insulin-like growth factor binding proteins IGFBP1/BP2; plasma GH increased at 1 year. Experimental validation indicates (1) hepatic GHR expression decreases in post-bariatric rats; (2) GHR knockdown in primary hepatocytes decreases gluconeogenic gene expression and glucose production. Thus, RYGB may induce resistance to diabetogenic effects of GH signaling. Trial Registration: Clinicaltrials.gov NCT01073020., Factors underlying the effects of gastric bypass surgery on glucose homeostasis are incompletely understood. Here the authors developed and applied high-throughput mediation analysis to identify proteome/metabolome mediators of improved glucose homeostasis after to gastric bypass surgery, and report that improved glycemia was mediated by the growth hormone receptor.
- Published
- 2020
21. A carnosine analog mitigates metabolic disorders of obesity by reducing carbonyl stress
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Giulio Vistoli, Lalage A. Katunga, Mara Colzani, Marina Carini, Luca Regazzoni, Stefania Gagliardi, Giuseppe Rossoni, Ettore Gilardoni, Danilo De Maddis, Giancarlo Aldini, Katsuhiko Funai, Ethan J. Anderson, Luca Cannizzaro, Renato Canevotti, and T. Blake Monroe
- Subjects
Male ,0301 basic medicine ,Dipeptidases ,Anserine ,Carnosine ,030209 endocrinology & metabolism ,macromolecular substances ,Pharmacology ,medicine.disease_cause ,Diabetes Mellitus, Experimental ,Rats, Sprague-Dawley ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Insulin resistance ,Metabolic Diseases ,Diabetes mellitus ,medicine ,Animals ,Humans ,Obesity ,Muscle, Skeletal ,Aldehydes ,General Medicine ,medicine.disease ,Mice, Mutant Strains ,Rats ,Oxidative Stress ,030104 developmental biology ,chemistry ,Commentary ,Insulin Resistance ,Metabolic syndrome ,Steatohepatitis ,Oxidative stress ,Dyslipidemia ,Research Article - Abstract
Sugar- and lipid-derived aldehydes are reactive carbonyl species (RCS) frequently used as surrogate markers of oxidative stress in obesity. A pathogenic role for RCS in metabolic diseases of obesity remains controversial, however, partly because of their highly diffuse and broad reactivity and the lack of specific RCS-scavenging therapies. Naturally occurring histidine dipeptides (e.g., anserine and carnosine) show RCS reactivity, but their therapeutic potential in humans is limited by serum carnosinases. Here, we present the rational design, characterization, and pharmacological evaluation of carnosinol, i.e., (2S)-2-(3-amino propanoylamino)-3-(1H-imidazol-5-yl)propanol, a derivative of carnosine with high oral bioavailability that is resistant to carnosinases. Carnosinol displayed a suitable ADMET (absorption, distribution, metabolism, excretion, and toxicity) profile and was determined to have the greatest potency and selectivity toward α,β-unsaturated aldehydes (e.g., 4-hydroxynonenal, HNE, ACR) among all others reported thus far. In rodent models of diet-induced obesity and metabolic syndrome, carnosinol dose-dependently attenuated HNE adduct formation in liver and skeletal muscle, while simultaneously mitigating inflammation, dyslipidemia, insulin resistance, and steatohepatitis. These improvements in metabolic parameters with carnosinol were not due to changes in energy expenditure, physical activity, adiposity, or body weight. Collectively, our findings illustrate a pathogenic role for RCS in obesity-related metabolic disorders and provide validation for a promising new class of carbonyl-scavenging therapeutic compounds rationally derived from carnosine.
- Published
- 2018
22. Development of Low-Molecular Weight Collagen Peptide Complex with Glycosaminoglycan Components
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P. V. Shekhovtsov, T. I. Nikolaeva, V. V. Kaptsov, K. S. Laurinavichus, and M. V. Molchanov
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0106 biological sciences ,Dipeptidases ,Swine ,Type II collagen ,Chymopapain ,Matrix (biology) ,Aminopeptidases ,01 natural sciences ,General Biochemistry, Genetics and Molecular Biology ,Glycosaminoglycan ,03 medical and health sciences ,Hydrolysis ,Chondrocytes ,0302 clinical medicine ,Enzymatic hydrolysis ,Papain ,medicine ,Animals ,Dipeptidyl-Peptidases and Tripeptidyl-Peptidases ,Collagen Type II ,Glycosaminoglycans ,Molecular mass ,Hyaline cartilage ,Chemistry ,Cartilage ,Biological Transport ,General Medicine ,Molecular Weight ,Hyaline Cartilage ,medicine.anatomical_structure ,Biochemistry ,Cattle ,Muramidase ,Proteoglycans ,Peptides ,030217 neurology & neurosurgery ,010606 plant biology & botany - Abstract
Enzymatic hydrolysis of biopolymers of the cartilage tissue was studied for obtaining a complex of type II collagen peptides and glycosaminoglycan oligosaccharides. Hydrothermal hydrolysis in a high pressure homogenizer followed by enzymatic hydrolysis of the cartilage tissue biopolymers with proteolytic enzyme preparation Karipazim yielded a complex of collagen peptides and glycosaminoglycan oligosaccharides with molecular weights of 240-720 Da. Low molecular weight of the components increases their bioavailability. Entering into the cells (chondrocytes), low-molecular-weight peptides, disaccharides, and oligosaccharides as structural elements of the matrix can participate in the formation of fibrils of collagen and proteoglycans. Exogenous substances replenish deficient components of the matrix and/or their concentrations, affect the formation and strengthen the cartilage tissue. Thus, using cattle and porcine hyaline cartilages, we prepared a complex of biopolymers with lower molecular weights in comparison with previously developed nutraceuticals.
- Published
- 2018
23. DPP8 is a novel therapeutic target for multiple myeloma
- Author
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Ayumi Tatekoshi, Kotaro Arita, Paras Jawaid, Nam H. Dang, Noriaki Iwao, Takashi Kondo, Satoshi Iyama, Chikao Morimoto, Yusuke Kamihara, Miho Arai, Ryo Hatano, Mati Ur Rehman, Jun Murakami, Kohichi Takada, Akinori Wada, Kyo Noguchi, Tsutomu Sato, Kei Ohnuma, Ichiro Yasuda, and Sayaka Kajikawa
- Subjects
0301 basic medicine ,Programmed cell death ,Dipeptidases ,lcsh:Medicine ,Myeloma ,Apoptosis ,Antineoplastic Agents ,CD38 ,Article ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Mice, Inbred NOD ,Cell Line, Tumor ,Drug Discovery ,medicine ,Cytotoxic T cell ,Animals ,Humans ,Vildagliptin ,Protease Inhibitors ,lcsh:Science ,Multiple myeloma ,Multidisciplinary ,Chemistry ,lcsh:R ,Proteolytic enzymes ,medicine.disease ,030104 developmental biology ,Cell culture ,030220 oncology & carcinogenesis ,Cancer research ,lcsh:Q ,Female ,Multiple Myeloma ,medicine.drug - Abstract
Dipeptidyl peptidases (DPPs) are proteolytic enzymes that are ideal therapeutic targets in human diseases. Indeed, DPP4 inhibitors are widely used in clinical practice as anti-diabetic agents. In this paper, we show that DPP4 inhibitors also induced cell death in multiple human myeloma cells. Among five DPP4 inhibitors, only two of them, vildagliptin and saxagliptin, exhibited apparent cytotoxic effects on myeloma cell lines, without any difference in suppression of DPP4 activity. As these two DPP4 inhibitors are known to have off-target effects against DPP8/9, we employed the specific DPP8/9 inhibitor 1G244. 1G244 demonstrated anti-myeloma effects on several cell lines and CD138+ cells from patients as well as in murine xenograft model. Through siRNA silencing approach, we further confirmed that DPP8 but not DPP9 is a key molecule in inducing cell death induced by DPP8/9 inhibition. In fact, the expression of DPP8 in CD38+ cells from myeloma patients was higher than that of healthy volunteers. DPP8/9 inhibition induced apoptosis, as evidenced by activated form of PARP, caspases-3 and was suppressed by the pan-caspase inhibitor Z-VAD-FMK. Taken together, these results indicate that DPP8 is a novel therapeutic target for myeloma treatment.
- Published
- 2019
24. Anserine (beta-alanyl-3-methyl-L-histidine) improves neurovascular-unit dysfunction and spatial memory in aged AβPPswe/PSEN1dE9 Alzheimer’s-model mice
- Author
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Kota Enomoto, Jun Kaneko, Akiko Enya, Qiong Ding, and Tatsuhiro Hisatsune
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Aging ,Dipeptidases ,Anserine ,Carnosine ,lcsh:Medicine ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Alzheimer Disease ,Internal medicine ,medicine ,Presenilin-1 ,Elderly people ,Animals ,Humans ,Neutral ph ,lcsh:Science ,Episodic memory ,Spatial Memory ,Multidisciplinary ,Amyloid beta-Peptides ,Beta-Alanyl-3-Methyl-L-Histidine ,lcsh:R ,Brain ,Endothelial Cells ,Neurovascular bundle ,Disease Models, Animal ,030104 developmental biology ,Endocrinology ,Cerebral blood flow ,chemistry ,Immunology ,lcsh:Q ,Pericytes ,Neuroglia ,030217 neurology & neurosurgery - Abstract
Anserine/carnosine supplementation improves cerebral blood flow and verbal episodic memory in elderly people, as we previously reported. Anserine’s buffering activity is superior to that of carnosine at neutral pH. In human sera, carnosine but not anserine is rapidly cleaved by carnosinase, limiting its effectiveness. This study examined the effects of anserine on AβPPswe/PSEN1dE9 Alzheimer’s disease (AD) model mice over 18-months old, an age at which these mice exhibit detectable memory deficits. We found that 8 weeks of anserine treatment completely recovered the memory deficits, improved pericyte coverage on endothelial cells in the brain, and diminished chronic glial neuroinflammatory reactions in these mice. These results suggest that anserine (beta-alanyl-3-methyl-L-histidine) supplementation improved memory functions in AD-model mice by exerting a protective effect on the neurovascular units, which are composed of endothelial cells, pericytes, and supporting glial cells.
- Published
- 2017
25. The Effects of Hyperbaric Oxygen Treatment on Total Antioxidant Capacity and Prolidase Activity after Bile Duct Ligation in Rats
- Author
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Pinar Atukeren, Hayriye Erman, Burhan Aksu, Duygu Terzioglu, Lebriz Uslu, Suleyman Ayvaz, Gonul Simsek, Hafize Uzun, and Remise Gelisgen
- Subjects
Male ,Dipeptidases ,medicine.medical_specialty ,Antioxidant ,medicine.medical_treatment ,medicine.disease_cause ,digestive system ,Antioxidants ,Rats, Sprague-Dawley ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cholestasis ,Predictive Value of Tests ,Fibrosis ,Internal medicine ,Animals ,Humans ,Medicine ,Ligation ,Common Bile Duct ,chemistry.chemical_classification ,Hyperbaric Oxygenation ,business.industry ,Bile duct ligation ,medicine.disease ,N-Acetylneuraminic Acid ,Rats ,Sialic acid ,Oxygen ,Disease Models, Animal ,Oxidative Stress ,Antioxidant capacity ,Endocrinology ,Liver ,chemistry ,Biochemistry ,Spectrophotometry ,030220 oncology & carcinogenesis ,Thiol ,030211 gastroenterology & hepatology ,Surgery ,business ,Biomarkers ,Oxidative stress - Abstract
BACKGROUND Hyperbaric oxygen (HBO) therapy may improve cholestasis, increase hepatic regeneration, and decrease oxidative stress in liver. In this study, we aimed to investigate the effects of HBO therapy on hepatic oxidative stress parameters, such as total thiol groups (T-SH), protein carbonyl (PCO), and total antioxidant capacity (TAC) as well as the predictive value of the noninvasive biochemical marker, sialic acid (SA), and prolidase activity in bile duct ligation (BDL)-induced oxidative damage and fibrosis in rats. METHODS We divided 32 adult male Sprague Dawley rats into four groups: sham, sham + HBO, BDL, and BDL + HBO; each group contained eight animals. We placed the sham + HBO and BDL + HBO groups in an experimental hyperbaric chamber, in which we administered pure oxygen at 2.5 atmospheres for 90 min on 14 consecutive days. RESULTS The application of BDL significantly increased PCO levels and prolidase activity, and decreased T-SH and TAC levels. HBO significantly decreased PCO levels and prolidase activity and increased T-SH and TAC levels in the liver tissues. There was no significant difference in sialic acid levels between any groups. CONCLUSIONS These results indicate that HBO therapy has hepatoprotective effects on BDL-induced injury by decreasing PCO and prolidase activity and increasing antioxidant activities. We therefore suggest that HBO therapy may be useful after BDL-induced injury.
- Published
- 2016
26. DPP8 and DPP9 inhibition induces pro-caspase-1-dependent monocyte and macrophage pyroptosis
- Author
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Sarah E. Poplawski, David G. Sanford, Mitchell S. Wang, Eun Bin Go, Daniel A. Bachovchin, William W. Bachovchin, Yuxin Liu, Darren C. Johnson, Michael O Arciprete, Ramya Sridharan, Ashley J. Chui, Wengen Wu, Marian C. Okondo, Todd R. Golub, and Jack H. Lai
- Subjects
0301 basic medicine ,Dipeptidases ,Proteases ,Programmed cell death ,Serine Proteinase Inhibitors ,Molecular Conformation ,Caspase 1 ,Article ,Cell Line ,Serine ,Mice ,Structure-Activity Relationship ,03 medical and health sciences ,Pyroptosis ,medicine ,Animals ,Humans ,Macrophage ,Dipeptidyl-Peptidases and Tripeptidyl-Peptidases ,Molecular Biology ,Dose-Response Relationship, Drug ,Chemistry ,Macrophages ,Monocyte ,Dipeptides ,Cell Biology ,Boronic Acids ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Lytic cycle ,Leukocytes, Mononuclear - Abstract
Val-boroPro (Talabostat, PT-100), a nonselective inhibitor of post-proline cleaving serine proteases, stimulates mammalian immune systems through an unknown mechanism of action. Despite this lack of mechanistic understanding, Val-boroPro has attracted substantial interest as a potential anticancer agent, reaching phase 3 trials in humans. Here we show that Val-boroPro stimulates the immune system by triggering a proinflammatory form of cell death in monocytes and macrophages known as pyroptosis. We demonstrate that the inhibition of two serine proteases, DPP8 and DPP9, activates the pro-protein form of caspase-1 independent of the inflammasome adaptor ASC. Activated pro-caspase-1 does not efficiently process itself or IL-1β but does cleave and activate gasdermin D to induce pyroptosis. Mice lacking caspase-1 do not show immune stimulation after treatment with Val-boroPro. Our data identify what is to our knowledge the first small molecule that induces pyroptosis and reveals a new checkpoint that controls the activation of the innate immune system.
- Published
- 2016
27. Family-wide annotation of enzymatic pathways by parallel in vivo metabolomics
- Author
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Veronica L. Li, Jonathan Z. Long, Joon T. Kim, Stephanie M. Terrell, and Curt R. Fischer
- Subjects
Male ,Dipeptidases ,Spectrometry, Mass, Electrospray Ionization ,Clinical Biochemistry ,Computational biology ,Biology ,01 natural sciences ,Biochemistry ,Article ,Amidohydrolases ,chemistry.chemical_compound ,Mice ,Metabolomics ,In vivo ,Drug Discovery ,Animals ,Humans ,Molecular Biology ,Chromatography, High Pressure Liquid ,Pharmacology ,chemistry.chemical_classification ,Mice, Knockout ,Dipeptide ,010405 organic chemistry ,Catabolism ,Hydrolysis ,Dipeptides ,Recombinant Proteins ,0104 chemical sciences ,Up-Regulation ,Mice, Inbred C57BL ,Metabolic pathway ,Enzyme ,HEK293 Cells ,chemistry ,Liver ,Mutagenesis, Site-Directed ,Molecular Medicine ,ACY1 ,Homeostasis - Abstract
Enzymes catalyze fundamental biochemical reactions that control cellular and organismal homeostasis. Here we present an approach for de novo biochemical pathway discovery across entire enzyme families using parallel viral transduction in mice and untargeted liquid chromatography-mass spectrometry. Applying this method to the mammalian M20 peptidases uncovers both known pathways of amino acid metabolism as well as a previously unknown CNDP2-regulated pathway for threonyl dipeptide catabolism. Ablation of CNDP2 in mice elevates threonyl dipeptides across multiple tissues, establishing the physiologic relevance of our biochemical assignments. Taken together, these data underscore the utility of parallel in vivo metabolomics for the family-wide discovery of enzymatic pathways. Enzymes catalyze fundamental biochemical reactions that control cellular and organismal homeostasis. Here we present an approach for de novo biochemical pathway discovery across entire enzyme families using parallel viral transduction in mice and untargeted liquid chromatography-mass spectrometry. Applying this method to the mammalian M20 peptidases uncovers known pathways of amino acid metabolism mediated by ACY1 (hydrolysis of N-acetyl amino acids) and CNDP2 (hydrolysis of carnosine). We also uncover a previously unknown CNDP2-regulated pathway for threonyl dipeptide catabolism. Ablation of CNDP2 in mice elevates threonyl dipeptides across multiple tissues, establishing the physiologic relevance of our biochemical assignments. Taken together, these data underscore the utility of parallel in vivo metabolomics for the family-wide discovery of enzymatic pathways.
- Published
- 2019
28. CNDP2 Acts as an Activator for Human Ovarian Cancer Growth and Metastasis via the PI3K/AKT Pathway
- Author
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Ling P Zhao, Li Q Zhang, Hong S Lu, Hua Q Yang, Xian J Chen, Su Q Yang, and Ying Wang
- Subjects
Cancer Research ,Dipeptidases ,endocrine system diseases ,growth ,medicine.disease_cause ,Metastasis ,03 medical and health sciences ,Mice ,Phosphatidylinositol 3-Kinases ,0302 clinical medicine ,Cell Movement ,Cell Line, Tumor ,medicine ,Animals ,Humans ,metastasis ,Neoplasm Metastasis ,PI3K/AKT/mTOR pathway ,030304 developmental biology ,Cell Proliferation ,Neoplasm Staging ,Mice, Knockout ,Ovarian Neoplasms ,0303 health sciences ,Chemistry ,Activator (genetics) ,medicine.disease ,Xenograft Model Antitumor Assays ,Gene Expression Regulation, Neoplastic ,CNDP2 ,Cytosol ,Disease Models, Animal ,Cell Transformation, Neoplastic ,ovarian cancer ,Oncology ,030220 oncology & carcinogenesis ,Molecular mechanism ,Cancer research ,Female ,Original Article ,Ovarian cancer ,Carcinogenesis ,Proto-Oncogene Proteins c-akt ,Signal Transduction - Abstract
Introduction: The mechanism of tumorigenesis and metastasis of ovarian cancer has not yet been elucidated. This study aimed to investigate the role and molecular mechanism of cytosolic nonspecific dipeptidase 2 in tumorigenesis and metastasis. Methods: Cytosolic nonspecific dipeptidase 2 expression in human ovarian cancer tissues and cell lines was assessed with methyl thiazolyl tetrazolium (MTT), clone formation, and transwell assays performed to evaluate the ability of ovarian cancer cells to proliferate and migrate. Nude mice tumor formation experiments were also performed by subcutaneously injecting cells with stable cytosolic nonspecific dipeptidase 2 knockdown and control SKOV3 cells into BALB/c female nude mice to detect changes in PI3K/AKT pathway-related proteins by Western blotting. Results: Cytosolic nonspecific dipeptidase 2 was highly expressed in human ovarian cancer tissues, with its expression associated with pathological data, including ovarian cancer metastasis. A cytosolic nonspecific dipeptidase 2 stable knockdown or ectopic expression ovarian cancer cell model was established and demonstrated that cytosolic nonspecific dipeptidase 2 could promote the proliferation of ovarian cancer cells. Transwell cell migration and invasion assays confirmed that cytosolic nonspecific dipeptidase 2 enhanced cell metastasis in ovarian cancer. Furthermore, in vivo xenograft experiments demonstrated that cytosolic nonspecific dipeptidase 2 can promote the development and progression of ovarian cancer, increasing the expression of phosphorylated PI3K and AKT. Conclusions: Cytosolic nonspecific dipeptidase 2 promotes the occurrence and development of ovarian cancer through the PI3K/AKT signaling pathway.
- Published
- 2019
29. Verapamil and collagenase differentially affect collagen metabolism in experimental model of Peyronie's disease
- Author
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Tomasz Guszczyn, Jacek Karaszewski, Ilona Zaręba, Barbara Darewicz, and Jerzy Pałka
- Subjects
Male ,Dipeptidases ,genetic structures ,Cell Survival ,Penile Induration ,Connective tissue ,Inflammation ,Biology ,Models, Biological ,Cell Line ,03 medical and health sciences ,Cell Movement ,medicine ,Animals ,Humans ,Viability assay ,Fibroblast ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Wound Healing ,030306 microbiology ,Cell growth ,Cell migration ,Cell Biology ,DNA ,Fibroblasts ,Molecular biology ,eye diseases ,medicine.anatomical_structure ,Verapamil ,Collagenase ,Collagen ,medicine.symptom ,Wound healing ,medicine.drug - Abstract
Objectives Peyronie's disease (PD) is accompanied by remodelling of connective tissue into fibrotic plaque. Treatment of the inflammatory and fibrotic phases of the disease is not established. The aim of the study was to evaluate the effect of verapamil (VER) and bacterial collagenase (COLL) on collagen metabolism and cell migration in fibroblasts with experimental wound healing and inflammation as an in vitro model of PD. Materials and methods In vitro model of PD was designed using experimental model of inflammation induced by Interleukin-1 (IL-1) in cultured fibroblasts and mechanical damage of the cells. Cell viability, cell proliferation, collagen biosynthesis, prolidase activity and cell migration were studied in both models of the cells treated with VER and COLL. Results VER decreased cell viability, DNA and collagen biosynthesis and increased prolidase activity in control fibroblast, while in “wounded” fibroblasts it significantly decreased all the processes. COLL did not affect cell viability and DNA biosynthesis, while inhibited collagen biosynthesis and prolidase activity in both control and “wounded” fibroblasts. In IL-1-treated fibroblasts VER inhibited all studied processes except prolidase activity, while COLL inhibited only collagen biosynthesis and prolidase activity. COLL accelerated cell migration, while VER attenuated the process in fibroblast model of wound healing, compared to control cells. Conclusion VER and COLL attenuate collagen biosynthesis in both fibroblast models. The VER-dependent inhibition of collagen biosynthesis was accompanied by inhibition of DNA biosynthesis at high prolidase activity, while COLL affected this process through inhibition of prolidase activity at high rate of DNA biosynthesis. It shows that anti-fibrotic activity of VER/COLL and anti-inflammatory activity of VER may represent approach to establish standard treatment of PD.
- Published
- 2019
30. Roles of cysteinyl leukotrienes and their receptors in immune cell-related functions
- Author
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Yoshihide, Kanaoka and K Frank, Austen
- Subjects
Inflammation ,Leukotriene E4 ,Receptors, Leukotriene ,Dipeptidases ,Leukotrienes ,Arachidonate 5-Lipoxygenase ,Group IV Phospholipases A2 ,5-Lipoxygenase-Activating Proteins ,Leukotriene C4 ,Mice ,Neoplasms ,Animals ,Humans ,Asthma, Aspirin-Induced ,Cysteine ,Glutathione Transferase - Abstract
The cysteinyl leukotrienes (cys-LTs), leukotriene C
- Published
- 2019
31. The testis-specifically expressed Dpep3 is not essential for male fertility in mice
- Author
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Qinghua Shi, Qamar Zaman, Xiaohua Jiang, Mazhar Khan, Fazal Wahab, Yubin Xie, Hanwei Jiang, Asim Ali, Hafiz Muhammad Jafar Hussain, Jianze Xu, Ranjha Khan, and Hui Ma
- Subjects
0301 basic medicine ,Male ,Dipeptidases ,Biology ,Andrology ,03 medical and health sciences ,Meiotic Prophase I ,Gene Knockout Techniques ,Mice ,0302 clinical medicine ,Gene duplication ,Testis ,Genetics ,Animals ,Humans ,Spermatogenesis ,Gene ,Conserved Sequence ,Infertility, Male ,Phylogeny ,Messenger RNA ,Sperm Count ,Membrane Proteins ,Histology ,General Medicine ,Sperm ,030104 developmental biology ,Organ Specificity ,030220 oncology & carcinogenesis ,Knockout mouse ,Sperm Motility - Abstract
More than 2300 genes have been reported to be involved in spermatogenesis but the functional roles of most genes in male fertility remain to be elucidated. In this study, we explored the function of dipeptidase 3 (Dpep3), a gene predicted to be testis-specific, in male fertility of mice. We showed that Dpep3 is evolutionarily conserved in human and mouse along with other eutherians. Its mRNA was exclusively detected in testicular tissue and expressed in testes from 7 days postpartum. To further explore its role in male fertility, we generated Dpep3 knockout mice (Dpep3−/−) using the CRISPR/Cas9 technology and found that the male Dpep3−/− mice are fertile despite a significant reduction in sperm count. Histology of testis and progression of meiotic prophase I showed no obvious difference between wild-type and Dpep3−/− mice. All these findings indicate that Dpep3 is not essential for male fertility in mice. These findings will help other researchers to avoid research duplication, save their time and resources to focus on the genes that are indispensable for male fertility.
- Published
- 2019
32. Dpep2 Emerging as a Modulator of Macrophage Inflammation Confers Protection Against CVB3-Induced Viral Myocarditis
- Author
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Sidong Xiong, Yan Yue, and Xiaoli Yang
- Subjects
0301 basic medicine ,Microbiology (medical) ,Dipeptidase ,Dipeptidases ,Viral Myocarditis ,030106 microbiology ,Immunology ,lcsh:QR1-502 ,Coxsackievirus Infections ,Inflammation ,Stimulation ,Microbiology ,lcsh:Microbiology ,law.invention ,03 medical and health sciences ,Cellular and Infection Microbiology ,law ,medicine ,Animals ,Immunologic Factors ,Macrophage ,Psychological repression ,Original Research ,Mice, Knockout ,Mice, Inbred BALB C ,biology ,viral myocarditis (VMC) ,business.industry ,Membrane Proteins ,coxsackievirus B3 (CVB3) ,Enterovirus B, Human ,macrophages ,Disease Models, Animal ,Myocarditis ,030104 developmental biology ,Infectious Diseases ,Dpep2 ,NF-κB signaling ,biology.protein ,cardiovascular system ,Suppressor ,medicine.symptom ,Signal transduction ,business - Abstract
Overwhelming cardiac inflammation has been reported to be the pathogenic mechanism of Coxsackievirus B3 (CVB3)-induced viral myocarditis (VMC), while the detailed molecular mechanisms remain unknown. Membrane-bound dipeptidases (MBD, also known as Dpep) have been shown to be involved in inflammatory diseases. However, the clear and direct evidence of their impacts on inflammation is still lacking. In this study, our results revealed that Dpep2 expression was remarkably increased during CVB3 infection, and primarily produced by the cardiac tissue-infiltrating macrophages instead of constitutive cardiomyocytes. Macrophages have been reported to play an important pathological role in driving VMC. Interestingly, macrophage-specific Dpep2 deletion robustly aggravated CVB3-induced cardiac inflammation, evidenced by augmented expression of TNF-α, IL-6, and MCP-1 in heart tissue. In addition, Dpep2-deficient bone-marrow derived macrophages (BMDMs) generated more TNF-α, IL-6, and MCP-1 after CVB3 stimulation compared with the control BMDMs. Moreover, this suppressive effect of Dpep2 on macrophages relied on its repression on NF-κB signaling pathway, but not on its conventional hydrolysate LTE4. Taken together, this study revealed that Dpep2 could protect against CVB3-induced VMC by acting as a suppressor of macrophage inflammation. Better understanding how macrophage Dpep2 dampened the cardiac inflammation would provide us with insights for the efficient control of CVB3-induced VMC.
- Published
- 2019
33. Prolidase enzyme is required for extracellular matrix integrity and impacts on postnatal cerebellar cortex development
- Author
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Roberta Besio, Marco Cocchia, Beatrice Ferrari, Maria Grazia Bottone, Federica Coppa, Graziella Bernocchi, Silvia Maruelli, Violetta Insolia, Erica Cecilia Priori, Antonella Forlino, Caterina Gasperini, and Erika Iervasi
- Subjects
0301 basic medicine ,Dipeptidases ,Synaptogenesis ,Cortical granule ,Fluorescent Antibody Technique ,Mice, Transgenic ,Biology ,Extracellular matrix ,03 medical and health sciences ,Cerebellar Cortex ,Mice ,0302 clinical medicine ,medicine ,Animals ,Basement membrane ,Mice, Inbred C3H ,Prolidase deficiency ,Cell growth ,General Neuroscience ,Cortical dysplasia ,medicine.disease ,Cell biology ,Extracellular Matrix ,030104 developmental biology ,medicine.anatomical_structure ,Animals, Newborn ,Cerebellar cortex ,Mice, Inbred CBA ,030217 neurology & neurosurgery - Abstract
The extracellular matrix is essential for brain development, lamination, and synaptogenesis. In particular, the basement membrane below the pial meninx (pBM) is required for correct cortical development. The last step in the catabolism of the most abundant protein in pBM, collagen Type IV, requires prolidase, an exopeptidase cleaving the imidodipeptides containing pro or hyp at the C-terminal end. Mutations impairing prolidase activity lead in humans to the rare disease prolidase deficiency characterized by severe skin ulcers and mental impairment. Thus, the dark-like (dal) mouse, in which the prolidase is knocked-out, was used to investigate whether the deficiency of prolidase affects the neuronal maturation during development of a brain cortex area. Focusing on the cerebellar cortex, thinner collagen fibers and disorganized pBM were found. Aberrant cortical granule cell proliferation and migration occurred, associated to defects in brain lamination, and in particular in maturation of Purkinje neurons and formation of synaptic contacts. This study deeply elucidates a link between prolidase activity and neuronal maturation shedding new light on the molecular basis of functional aspects in the prolidase deficiency.
- Published
- 2019
34. Focusing on the functional characterization of the anserinase from Oreochromis niloticus
- Author
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Francesco Bellia, E. Rizzarelli, Luciano Pirone, S. Di Gaetano, and Emilia Pedone
- Subjects
Dipeptidase ,Dipeptidases ,Anserine ,Carnosine ,02 engineering and technology ,medicine.disease_cause ,Biochemistry ,law.invention ,Substrate Specificity ,03 medical and health sciences ,chemistry.chemical_compound ,Protein sequencing ,Structural Biology ,law ,medicine ,Animals ,Amino Acid Sequence ,Cloning, Molecular ,Molecular Biology ,Escherichia coli ,Histidine ,030304 developmental biology ,Cloning ,0303 health sciences ,biology ,General Medicine ,Cichlids ,Dipeptides ,021001 nanoscience & nanotechnology ,chemistry ,Recombinant DNA ,biology.protein ,Transition metal ions ,0210 nano-technology - Abstract
Carnosine, anserine and homocarnosine are the three most representative compounds of the histidine dipeptides family, widely distributed in mammals in different amounts depending on the species and the tissue considered. Histidine dipeptides are mainly degraded by two different carnosinase homologues: a highly specific metal-ion dependent carnosinase (CN1) located in serum and brain and a non-specific cytosolic form (CN2). The hydrolysis of such dipeptides in prokaryotes and eukaryotes is also catalyzed by the anserinase (ANSN). Such naturally occurring dipeptides represent an interesting topic because they seem to have numerous biological roles such as potential neuroprotective and neurotransmitter functions in the brain and therefore ANSN results to be a very interesting target of study. We here report, for the first time, cloning, expression of ANSN from the fish Oreochromis niloticus both in a mammalian and in a prokaryotic system, in order to perform deep functional studies by enzymatic assays in the presence of different metals and substrates. Furthermore, by means of a mass spectrometry-based proteomic approach, we analysed protein sequence and the potential presence of post-translational modifications in the mammalian recombinant protein. Finally, a preliminary structural characterization was carried out on ANSN produced in Escherichia coli.
- Published
- 2019
35. CNDP1 knockout in zebrafish alters the amino acid metabolism, restrains weight gain, but does not protect from diabetic complications
- Author
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Jakob Morgenstern, Michael Büttner, Tim Weigand, Verena Peters, Gernot Poschet, Peter P. Nawroth, Felix Schmöhl, Nadine Volk, Claus Peter Schmitt, Jens Kroll, Tanja Poth, Xiaogang Li, and Thomas Fleming
- Subjects
medicine.medical_specialty ,Dipeptidases ,Carnosine ,Kidney ,Weight Gain ,Diabetic nephropathy ,Diabetes Complications ,Cellular and Molecular Neuroscience ,chemistry.chemical_compound ,Gene Knockout Techniques ,Internal medicine ,Diabetes mellitus ,medicine ,Animals ,Amino Acids ,Molecular Biology ,Zebrafish ,Pharmacology ,chemistry.chemical_classification ,biology ,Cell Biology ,Metabolism ,biology.organism_classification ,medicine.disease ,Amino acid ,Enzyme ,Endocrinology ,chemistry ,Diabetes Mellitus, Type 2 ,Molecular Medicine ,Intracellular - Abstract
The gene CNDP1 was associated with the development of diabetic nephropathy. Its enzyme carnosinase 1 (CN1) primarily hydrolyzes the histidine-containing dipeptide carnosine but other organ and metabolic functions are mainly unknown. In our study we generated CNDP1 knockout zebrafish, which showed strongly decreased CN1 activity and increased intracellular carnosine levels. Vasculature and kidneys of CNDP1-/- zebrafish were not affected, except for a transient glomerular alteration. Amino acid profiling showed a decrease of certain amino acids in CNDP1-/- zebrafish, suggesting a specific function for CN1 in the amino acid metabolisms. Indeed, we identified a CN1 activity for Ala-His and Ser-His. Under diabetic conditions increased carnosine levels in CNDP1-/- embryos could not protect from respective organ alterations. Although, weight gain through overfeeding was restrained by CNDP1 loss. Together, zebrafish exhibits CN1 functions, while CNDP1 knockout alters the amino acid metabolism, attenuates weight gain but cannot protect organs from diabetic complications.
- Published
- 2018
36. A Global Cndp1-Knock-Out Selectively Increases Renal Carnosine and Anserine Concentrations in an Age- and Gender-Specific Manner in Mice
- Author
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Tim Weigand, Florian Colbatzky, Tilman Pfeffer, Sven F. Garbade, Kristina Klingbeil, Michael Becker, Johanna Zemva, Ruben Bulkescher, Robin Schürfeld, Christian Thiel, Nadine Volk, David Reuss, Georg F. Hoffmann, Marc Freichel, Markus Hecker, Tanja Poth, Thomas Fleming, Gernot Poschet, Claus P. Schmitt, and Verena Peters
- Subjects
Blood Glucose ,Male ,0301 basic medicine ,Dipeptidases ,Arginine ,Cndp1 ,Carnosine ,lcsh:Chemistry ,Diabetic nephropathy ,Mice ,chemistry.chemical_compound ,0302 clinical medicine ,Insulin ,Glucose homeostasis ,CN1 ,Diabetic Nephropathies ,Amino Acids ,lcsh:QH301-705.5 ,Spectroscopy ,Mice, Knockout ,Kidney ,General Medicine ,Computer Science Applications ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Female ,kidney ,medicine.medical_specialty ,anserine ,Renal cortex ,Anserine ,Article ,Catalysis ,Inorganic Chemistry ,03 medical and health sciences ,Internal medicine ,medicine ,glucose homeostasis ,Animals ,HSP70 Heat-Shock Proteins ,RNA, Messenger ,Physical and Theoretical Chemistry ,Molecular Biology ,Organic Chemistry ,medicine.disease ,Mice, Inbred C57BL ,Glutamine ,Glucose ,030104 developmental biology ,Endocrinology ,lcsh:Biology (General) ,lcsh:QD1-999 ,chemistry ,carnosinase 1 - Abstract
Carnosinase 1 (CN1) is encoded by the Cndp1 gene and degrades carnosine and anserine, two natural histidine-containing dipeptides. In vitro and in vivo studies suggest carnosine- and anserine-mediated protection against long-term sequelae of reactive metabolites accumulating, e.g., in diabetes mellitus. We have characterized the metabolic impact of CN1 in 11- and 55-week-old Cndp1-knockout (Cndp1-KO) mice and litter-matched wildtypes (WT). In Cndp1-KO mice, renal carnosine and anserine concentrations were gender-specifically increased 2- to 9-fold, respectively in the kidney and both most abundant in the renal cortex, but remained unchanged in all other organs and in serum. Renal oxidized/reduced glutathione concentrations, renal morphology and function were unaltered. In Cndp1-KO mice at week 11, renal asparagine, serine and glutamine levels and at week 55, renal arginine concentration were reduced. Renal heat-shock-protein 70 (Hspa1a/b) mRNA declined with age in WT but not in Cndp1-KO mice, transcription factor heat-shock-factor 1 was higher in 55-week-old KO mice. Fasting blood glucose concentrations decreased with age in WT mice, but were unchanged in Cndp1-KO mice. Blood glucose response to intraperitoneal insulin was gender- but not genotype-dependent, the response to intraperitoneal glucose injection was similar in all groups. A global Cndp1-KO selectively, age- and gender-specifically, increases renal carnosine and anserine concentrations, alters renal amino acid- and HSP70 profile and modifies systemic glucose homeostasis. Increase of the natural occurring carnosine and anserine levels in the kidney by modulation of CN1 represents a promising therapeutic approach to mitigate or prevent chronic kidney diseases such as diabetic nephropathy.
- Published
- 2020
37. Pivotal role of carnosine in the modulation of brain cells activity: Multimodal mechanism of action and therapeutic potential in neurodegenerative disorders
- Author
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Filippo Caraci, Giuseppe Caruso, and Renaud Jolivet
- Subjects
0301 basic medicine ,Dipeptidases ,Carnosinemia ,Cognitive decline ,Carnosine ,Context (language use) ,Nitric oxide ,Brain metabolism ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Detoxification ,Glial cells ,medicine ,Animals ,Humans ,Amino Acid Metabolism, Inborn Errors ,Alzheimer’s disease ,Neurodegenerative disorders ,business.industry ,General Neuroscience ,Metabolic disorder ,Brain ,Brain Diseases, Metabolic, Inborn ,Neurodegenerative Diseases ,medicine.disease ,3. Good health ,030104 developmental biology ,Mechanism of action ,chemistry ,medicine.symptom ,business ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Carnosine (β-alanyl-l-histidine), a dipeptide, is an endogenous antioxidant widely distributed in excitable tissues like muscles and the brain. Although discovered more than a hundred years ago and having been extensively studied in the periphery, the role of carnosine in the brain remains mysterious. Carnosinemia, a rare metabolic disorder with increased levels of carnosine in urine and low levels or absence of carnosinase in the blood, is associated with severe neurological symptoms in humans. This review deals with the role of carnosine in the brain in both physiological and pathological conditions, with a focus on preclinical evidence suggesting a high therapeutic potential of carnosine in neurodegenerative disorders. We review carnosine and carnosinemia's discoveries and the extensive research on the role and benefits of carnosine in the periphery. We then turn to carnosine's biochemistry and distribution in the brain. Using an array of recent observations as a foundation, we draw a parallel with the role of carnosine in muscles and speculate on the role of carnosine in promoting the metabolic support of neurons by glial cells. Finally, carnosine has been shown to exert a multimodal activity including inhibition of protein cross-linking and aggregation of amyloid-β and related proteins, free radical generation, nitric oxide detoxification, and an anti-inflammatory activity. It could thus play an important role in the prevention and treatment of neurodegenerative diseases such as Alzheimer's disease. We discuss the potential of carnosine in this context and speculate on new preclinical research directions.
- Published
- 2018
38. Special Sections for the 6th ASM Conference on Cell-Cell Communication in Bacteria
- Author
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George A. O'Toole
- Subjects
Peptidoglycan metabolism ,Dipeptidases ,Cell signaling ,Mycobacterium smegmatis ,Cell ,Peptidoglycan ,Microbiology ,Gene Expression Regulation, Enzymologic ,Mice ,medicine ,Animals ,Molecular Biology ,RAW 264.7 Cells ,Regulation of gene expression ,biology ,Biofilm ,Gene Expression Regulation, Bacterial ,biology.organism_classification ,Coculture Techniques ,Anti-Bacterial Agents ,Cell biology ,Editorial ,medicine.anatomical_structure ,Biofilms ,Coculture Technique ,Gene Deletion ,Bacteria - Published
- 2018
39. Two <scp>dd</scp> -Carboxypeptidases from Mycobacterium smegmatis Affect Cell Surface Properties through Regulation of Peptidoglycan Cross-Linking and Glycopeptidolipids
- Author
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Anindya S. Ghosh, Ankita Bansal, Ganesh Kumar N, Satya Deo Pandey, Sathi Mallick, and Shilpa Pal
- Subjects
0301 basic medicine ,Dipeptidases ,Glycan ,Mycobacterium smegmatis ,030106 microbiology ,Mutant ,Peptidoglycan ,Cell morphology ,Microbiology ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,Bacterial Proteins ,Animals ,Molecular Biology ,biology ,Macrophages ,Genetic Complementation Test ,Biofilm ,Gene Expression Regulation, Bacterial ,biology.organism_classification ,Phenotype ,Anti-Bacterial Agents ,Complementation ,RAW 264.7 Cells ,chemistry ,Biochemistry ,biology.protein ,Glycolipids ,Research Article - Abstract
During the peptidoglycan (PG) maturation of mycobacteria, the glycan strands are interlinked by both 3-3 (between two meso -diaminopimelic acids [ meso -DAPs]) and 4-3 cross-links (between d -Ala and meso -DAP), though there is a predominance (60 to 80%) of 3-3 cross-links. The dd -carboxypeptidases ( dd -CPases) act on pentapeptides to generate tetrapeptides that are used by ld -transpeptidases as substrates to form 3-3 cross-links. Therefore, dd -CPases play a crucial role in mycobacterial PG cross-link formation. However, the physiology of dd -CPases in mycobacteria is relatively unexplored. In this study, we deleted two dd -CPase genes, msmeg_2433 and msmeg_2432 , both individually and in combination, from Mycobacterium smegmatis mc 2 155. Though the single dd -CPase gene deletions had no significant impact on the mycobacterial physiology, many interesting functional alterations were observed in the double-deletion mutant, viz ., a predominance in PG cross-link formation was shifted from 3-3 cross-links to 4-3, cell surface glycopeptidolipid (GPL) expression was reduced, and susceptibility to β-lactams and antitubercular agents was enhanced. Moreover, the survival rate of the double mutant within murine macrophages was higher than that of the parent. Interestingly, the complementation with any one of the dd -CPase genes could restore the wild-type phenotype. In a nutshell, we infer that the altered ratio of 4-3 to 3-3 PG cross-links might have influenced the expression of surface GPLs, colony morphology, biofilm formation, drug susceptibility, and subsistence of the cells within macrophages. IMPORTANCE The glycan strands in mycobacterial peptidoglycan (PG) are interlinked by both 3-3 and 4-3 cross-links. The dd -CPases generate tetrapeptides by acting on the pentapeptides, and ld -transpeptidases use tetrapeptides as substrates to form 3-3 cross-links. In this study, we showed that simultaneous deletions of two dd -CPases alter the nature of PG cross-linking from 3-3 cross-links to 4-3 cross-links. The deletions subsequently decrease the expression of glycopeptidolipids (significant surface lipid present in many nontuberculous mycobacteria, including Mycobacterium smegmatis ) and affect other physiological parameters, like cell morphology, growth rate, biofilm formation, antibiotic susceptibility, and survival within murine macrophages. Thus, unraveling the physiology of dd -CPases might help us design antimycobacterial therapeutics in the future.
- Published
- 2018
40. Kidney surveillance in the spotlight: contrast-induced acute kidney injury illuminated
- Author
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Simon J. Atkinson
- Subjects
0301 basic medicine ,Pathology ,medicine.medical_specialty ,Dipeptidases ,media_common.quotation_subject ,Renal function ,Contrast Media ,Kidney ,urologic and male genital diseases ,03 medical and health sciences ,Mice ,medicine ,Contrast (vision) ,Animals ,Humans ,media_common ,Heterogeneous group ,business.industry ,urogenital system ,Acute kidney injury ,General Medicine ,Acute Kidney Injury ,medicine.disease ,Immune surveillance ,030104 developmental biology ,Innovative Therapies ,medicine.anatomical_structure ,Commentary ,business ,Research Article - Abstract
Radiographic contrast agents cause acute kidney injury (AKI), yet the underlying pathogenesis is poorly understood. Nod-like receptor pyrin containing 3–deficient (Nlrp3-deficient) mice displayed reduced epithelial cell injury and inflammation in the kidney in a model of contrast-induced AKI (CI-AKI). Unexpectedly, contrast agents directly induced tubular epithelial cell death in vitro that was not dependent on Nlrp3. Rather, contrast agents activated the canonical Nlrp3 inflammasome in macrophages. Intravital microscopy revealed diatrizoate (DTA) uptake within minutes in perivascular CX3CR1+ resident phagocytes in the kidney. Following rapid filtration into the tubular luminal space, DTA was reabsorbed and concentrated in tubular epithelial cells via the brush border enzyme dipeptidase-1 in volume-depleted but not euvolemic mice. LysM-GFP+ macrophages recruited to the kidney interstitial space ingested contrast material transported from the urine via direct interactions with tubules. CI-AKI was dependent on resident renal phagocytes, IL-1, leukocyte recruitment, and dipeptidase-1. Levels of the inflammasome-related urinary biomarkers IL-18 and caspase-1 were increased immediately following contrast administration in patients undergoing coronary angiography, consistent with the acute renal effects observed in mice. Taken together, these data show that CI-AKI is a multistep process that involves immune surveillance by resident and infiltrating renal phagocytes, Nlrp3-dependent inflammation, and the tubular reabsorption of contrast via dipeptidase-1.
- Published
- 2018
41. Rectal application of argan oil improves healing of colorectal anastomosis in rats1
- Author
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Aziz Mutlu, Barlas, Serdar, Kuru, Kemal, Kismet, Turgut, Cavusoglu, Yusuf Murat, Bag, Mehmet, Senes, Neslihan, Cihan, Pinar, Celepli, Yilmaz, Unal, and Sema, Hucumenoglu
- Subjects
Dipeptidases ,Wound Healing ,Colon ,Anastomosis, Surgical ,Surgical Wound ,Anti-Inflammatory Agents ,Rectum ,Reproducibility of Results ,Antioxidants ,Hydroxyproline ,Oxidative Stress ,Random Allocation ,Treatment Outcome ,Spectrophotometry ,Malondialdehyde ,Animals ,Plant Oils ,Female ,Collagen ,Rats, Wistar ,Oxidoreductases - Abstract
To investigate the possible effects of argan oil on the healing of colorectal anastomoses.n Group 1 (sham), laparotomy was performed and the colon was mobilized. In the control (Group 2) and argan oil (Group 3) groups, colonic resection and anastomosis were applied. To the control and sham groups, 2 mL of 0.9% NaCl was administred rectally, and in the argan oil group, 2 mL/day argan oil was applied rectally for 7 days.The mean bursting pressures of the argan oil and sham groups were significantly higher than the values in the control group. A significant difference was determined between the tissue hydroxyproline and prolidase levels of control group and other groups. Histopathologically, argan oil showed significant beneficial effects on colonic wound healing. In the argan oil and sham groups, the tissue malondialdehyde and fluorescent oxidation product levels were found to be lower and total sulfhydryl levels were higher than the control group.The rectally administered argan oil was observed to have significantly ameliorated wound healing parameters and exerted a significant antioxidant effect. This is the first study in the literature about the beneficial effects of argan oil on colorectal anastomoses.
- Published
- 2018
42. Identification and characterisation of carnostatine (SAN9812), a potent and selective carnosinase (CN1) inhibitor with in vivo activity
- Author
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Shiqi Zhang, Diego O. Pastene, Michael Teufel, Angelica Rodriguez-Niño, Jiedong Qiu, Hans-Peter Hammes, Sibylle J. Hauske, Bernhard K. Krämer, Thomas Albrecht, Sven Ruf, Aimo Kannt, Jacob van den Born, Harry van Goor, Verena Peters, Markus Kohlmann, Benito A. Yard, Groningen Institute for Organ Transplantation (GIOT), and Groningen Kidney Center (GKC)
- Subjects
0301 basic medicine ,Male ,Dipeptidases ,Clinical Biochemistry ,Carnosine ,Gene Expression ,Pharmacology ,Biochemistry ,SUPPLEMENTATION ,law.invention ,Diabetic nephropathy ,chemistry.chemical_compound ,Mice ,law ,Drug Discovery ,Transgenes ,Kidney ,Aminoacyl-histidine dipeptidase ,STAGE RENAL-DISEASE ,Imidazoles ,Translation (biology) ,Highly selective ,Recombinant Proteins ,medicine.anatomical_structure ,Recombinant DNA ,SECRETION ,Female ,LEUCINE REPEAT ,Protein Binding ,Adult ,CNDP1 GENOTYPE ,Transgene ,Injections, Subcutaneous ,Mice, Transgenic ,DIABETIC-NEPHROPATHY ,Small Molecule Libraries ,03 medical and health sciences ,In vivo ,Diabetic nephropathies ,medicine ,Animals ,Humans ,Protease Inhibitors ,MUSCLE CARNOSINE ,030102 biochemistry & molecular biology ,Organic Chemistry ,PERFORMANCE ,medicine.disease ,GENE ,POLYMORPHISM ,Mice, Inbred C57BL ,Kinetics ,030104 developmental biology ,chemistry ,Propionates - Abstract
Carnosinase 1 (CN1) has been postulated to be a susceptibility factor for developing diabetic nephropathy (DN). Although its major substrate, carnosine, is beneficial in rodent models of DN, translation of these findings to humans has been hampered by high CN1 activity in human serum resulting in rapid degradation of carnosine. To overcome this hurdle, we screened a protease-directed small-molecule library for inhibitors of human recombinant CN1. We identified SAN9812 as a potent and highly selective inhibitor of CN1 activity with a K-i of 11nM. It also inhibited CN1 activity in human serum and serum of transgenic mice-overexpressing human CN1. Subcutaneous administration of 30mg/kg SAN9812 led to a sustained reduction in circulating CN1 activity in human CN1 transgenic (TG) mice. Simultaneous administration of carnosine and SAN9812 increased carnosine levels in plasma and kidney by up to 100-fold compared to treatment-naive CN1-overexpressing mice. To our knowledge, this is the first study reporting on a potent and selective CN1 inhibitor with in vivo activity. SAN9812, also called carnostatine, may be used to increase renal carnosine concentration as a potential therapeutic modality for renal diseases linked to glycoxidative conditions.
- Published
- 2018
43. FL-926-16, a novel bioavailable carnosinase-resistant carnosine derivative, prevents onset and stops progression of diabetic nephropathy in db/db mice
- Author
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Iacobini, C., Menini, S., Blasetti Fantauzzi, C., Pesce, C. M., Giaccari, A., Salomone, E., Lapolla, A., Orioli, M., Aldini, G., and Pugliese, G.
- Subjects
Male ,Dipeptidases ,Animals ,Biological Availability ,Carnosine ,Cells, Cultured ,Diabetic Nephropathies ,Mice ,Mice, Inbred C57BL ,Mice, Transgenic ,Random Allocation ,Disease Progression ,renal injury ,Cells ,design ,receptor-mediated mechanisms ,Inbred C57BL ,Transgenic ,lipid-peroxidation ,induced glomerular injury ,glycation end-products ,apoe-null mice ,kidney-disease ,atherosclerosis ,accumulation ,Pharmacology ,Cultured ,Settore MED/13 - ENDOCRINOLOGIA ,Research Papers ,Nephropathy - Abstract
The advanced glycation end products (AGEs) participate in the pathogenesis of diabetic nephropathy (DN) by promoting renal inflammation and injury. L-carnosine acts as a quencher of the AGE precursors reactive carbonyl species (RCS), but is rapidly inactivated by carnosinase. In this study, we evaluated the effect of FL-926-16, a carnosinase-resistant and bioavailable carnosine derivative, on the onset and progression of DN in db/db mice.Adult male db/db mice and coeval db/m controls were left untreated or treated with FL-926-16 (30 mg·kgIn the prevention protocol, FL-926-16 significantly attenuated increases in creatinine (-80%), albuminuria (-77%), proteinuria (-75%), mean glomerular area (-34%), fractional (-40%) and mean (-42%) mesangial area in db/db mice. This protective effect was associated with a reduction in glomerular matrix protein expression and cell apoptosis, circulating and tissue oxidative and carbonyl stress, and renal inflammatory markers, including the NLRP3 inflammasome. In the regression protocol, the progression of DN was completely blocked, although not reversed, by FL-926-16. In cultured mesangial cells, FL-926-16 prevented NLRP3 expression induced by RCS but not by the AGE NFL-926-16 is effective at preventing the onset of DN and halting its progression in db/db mice by quenching RCS, thereby reducing the accumulation of their protein adducts and the consequent inflammatory response. In a future perspective, this novel compound may represent a promising AGE-reducing approach for DN therapy.
- Published
- 2018
44. Lack of prolidase causes a bone phenotype both in human and in mouse
- Author
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Sarah Foster, Antonio Rossi, Ersilia De Lorenzi, Isabella Villa, Ruggero Tenni, Roberta Gioia, Antonella Forlino, Silvia Maruelli, Halil Ibrahim Aydin, Maja Di Rocco, Josè Luis Dapena Diaz, Charu Deshpande, Çiğdem Seher Kasapkara, Teresa M. Gunn, Nick Bishop, Mehmet Ali Gürer, Roberta Besio, Raffaella Colombo, Haether Moore-Barton, Bartłomiej Kwiek, Peter Grabowski, Orla Gallagher, Ayşegül Tokatlı, Esra Adişen, and James M. Phang
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Male ,Dipeptidases ,Physiology ,Endocrinology, Diabetes and Metabolism ,Mice ,chemistry.chemical_compound ,Cytosol ,Mutant protein ,Body Size ,Femur ,Child ,Bone growth ,Mice, Inbred C3H ,Prolidase deficiency ,Protein catabolism ,Phenotype ,medicine.anatomical_structure ,Child, Preschool ,Female ,medicine.symptom ,Adult ,medicine.medical_specialty ,Histology ,Adolescent ,Molecular Sequence Data ,Mice, Transgenic ,Biology ,Short stature ,Bone and Bones ,Chondrocyte ,Young Adult ,Hydroxyproline ,Internal medicine ,medicine ,Animals ,Humans ,Retrospective Studies ,Osteoblasts ,Base Sequence ,Tibia ,Catabolism ,X-Ray Microtomography ,Fibroblasts ,medicine.disease ,Protein Structure, Tertiary ,Endocrinology ,chemistry ,Mice, Inbred CBA ,Prolidase Deficiency ,Tomography, X-Ray Computed - Abstract
The degradation of the main fibrillar collagens, collagens I and II, is a crucial process for skeletal development. The most abundant dipeptides generated from the catabolism of collagens contain proline and hydroxyproline. In humans, prolidase is the only enzyme able to hydrolyze dipeptides containing these amino acids at their C-terminal end, thus being a key player in collagen synthesis and turnover. Mutations in the prolidase gene cause prolidase deficiency (PD), a rare recessive disorder. Here we describe 12 PD patients, 9 of whom were molecularly characterized in this study. Following a retrospective analysis of all of them a skeletal phenotype associated with short stature, hypertelorism, nose abnormalities, microcephaly, osteopenia and genu valgum, independent of both the type of mutation and the presence of the mutant protein was identified. In order to understand the molecular basis of the bone phenotype associated with PD, we analyzed a recently identified mouse model for the disease, the dark-like (dal) mutant. The dal/dal mice showed a short snout, they were smaller than controls, their femurs were significantly shorter and pQCT and mu CT analyses of long bones revealed compromised bone properties at the cortical and at the trabecular level in both male and female animals. The differences were more pronounce at 1 month being the most parameters normalized by 2 months of age. A delay in the formation of the second ossification center was evident at postnatal day 10. Our work reveals that reduced bone growth was due to impaired chondrocyte proliferation and increased apoptosis rate in the proliferative zone associated with reduced hyperthrophic zone height. These data suggest that lack of prolidase, a cytosolic enzyme involved in the final stage of protein catabolism, is required for normal skeletogenesis especially at early age when the requirement for collagen synthesis and degradation is the highest. (C) 2014 Elsevier Inc. All rights reserved.
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- 2015
45. Investigation of possible prophylactic, renoprotective, and cardioprotective effects of thromboprophylactic drugs against ischemia–reperfusion injury
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Oguz Karahan, Ahmet Caliskan, Orhan Tezcan, Orkut Güçlü, Suleyman Yazici, İbrahim Kaplan, Celal Yavuz, and Sinan Demirtaş
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Male ,Dipeptidases ,Drug Evaluation, Preclinical ,Cardioprotection ,Pharmacology ,Kidney ,Rats, Sprague-Dawley ,chemistry.chemical_compound ,Rivaroxaban ,Malondialdehyde ,Medicine(all) ,lcsh:R5-920 ,Anticoagulant ,General Medicine ,Clopidogrel ,Hindlimb ,Femoral Artery ,Anticoagulant drugs ,Anesthesia ,Reperfusion Injury ,lcsh:Medicine (General) ,Enoxaparin sodium ,medicine.drug ,Cardiotonic Agents ,Ticlopidine ,medicine.drug_class ,Ischemia–reperfusion injury ,Morpholines ,Ischemia ,Thiophenes ,Nitric Oxide ,medicine ,Animals ,Enoxaparin ,Aspirin ,business.industry ,Aryldialkylphosphatase ,Myocardium ,Anticoagulants ,Renoprotection ,Heparin, Low-Molecular-Weight ,medicine.disease ,Antiaggregant drugs ,Oxidative Stress ,Bemiparin sodium ,chemistry ,business ,Reperfusion injury - Abstract
The aim of this study was to investigate whether anticoagulant and antiaggregant agents have protective effects against oxidative damage induced by peripheral ischemia–reperfusion (I/R). Groups were created as follows: control group, I/R group (sham group), I/R plus acetylsalicylic acid (Group I), I/R+clopidogrel (Group II), I/R+rivaroxaban (Group III), I/R+bemiparin sodium (Group IV), and I/R+enoxaparin sodium (Group V). In Groups I, II, III, IV, and V, drugs were administered daily for 1 week before I/R creation. Peripheral I/R was induced in the I/R groups by clamping the right femoral artery. The rats were sacrificed 1 hour after reperfusion. Nitrogen oxide levels, malondialdehyde (MDA) levels, paraoxonase-1 (PON1) activity, and prolidase activity were evaluated in both cardiac and renal tissues. There was no significant difference in nitrogen oxide levels between the groups. However, cardiac and renal MDA were significantly higher and PON1 activity was markedly lower in the I/R groups compared with the control group (p
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- 2015
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46. Renal immune surveillance and dipeptidase-1 contribute to contrast-induced acute kidney injury
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Craig N. Jenne, Bas G.J. Surewaard, Justin Chun, Arthur Lau, Paul Kubes, Jaye M. Platnich, Michelle C. Nelson, Matthew T. James, Anthony M. Jevnikar, Donna L. Senger, Saurav Roy Choudhury, Hyunjae Chung, Hallgrimur Benediktsson, Sarah L. Snelgrove, Daniel A. Muruve, Michael J. Hickey, Christina F. Sandall, Justin A. MacDonald, Takanori Komada, Victor Naumenko, Annegret Ulke-Lemée, and Paul L. Beck
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0301 basic medicine ,Nephrology ,Pathology ,medicine.medical_specialty ,Dipeptidases ,Inflammasomes ,Contrast Media ,Inflammation ,030204 cardiovascular system & hematology ,GPI-Linked Proteins ,Kidney ,Pathogenesis ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Internal medicine ,NLR Family, Pyrin Domain-Containing 3 Protein ,Leukocytes ,Medicine ,Animals ,Humans ,Immunologic Surveillance ,Mice, Knockout ,Phagocytes ,urogenital system ,business.industry ,Acute kidney injury ,Inflammasome ,General Medicine ,Acute Kidney Injury ,medicine.disease ,3. Good health ,Mice, Inbred C57BL ,Disease Models, Animal ,030104 developmental biology ,medicine.anatomical_structure ,Renal physiology ,medicine.symptom ,business ,Intravital microscopy ,medicine.drug - Abstract
Radiographic contrast agents cause acute kidney injury (AKI), yet the underlying pathogenesis is poorly understood. Nod-like receptor pyrin containing 3-deficient (Nlrp3-deficient) mice displayed reduced epithelial cell injury and inflammation in the kidney in a model of contrast-induced AKI (CI-AKI). Unexpectedly, contrast agents directly induced tubular epithelial cell death in vitro that was not dependent on Nlrp3. Rather, contrast agents activated the canonical Nlrp3 inflammasome in macrophages. Intravital microscopy revealed diatrizoate (DTA) uptake within minutes in perivascular CX3CR1+ resident phagocytes in the kidney. Following rapid filtration into the tubular luminal space, DTA was reabsorbed and concentrated in tubular epithelial cells via the brush border enzyme dipeptidase-1 in volume-depleted but not euvolemic mice. LysM-GFP+ macrophages recruited to the kidney interstitial space ingested contrast material transported from the urine via direct interactions with tubules. CI-AKI was dependent on resident renal phagocytes, IL-1, leukocyte recruitment, and dipeptidase-1. Levels of the inflammasome-related urinary biomarkers IL-18 and caspase-1 were increased immediately following contrast administration in patients undergoing coronary angiography, consistent with the acute renal effects observed in mice. Taken together, these data show that CI-AKI is a multistep process that involves immune surveillance by resident and infiltrating renal phagocytes, Nlrp3-dependent inflammation, and the tubular reabsorption of contrast via dipeptidase-1.
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- 2017
47. Carnosinase, diabetes mellitus and the potential relevance of carnosinase deficiency
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Verena Peters, Johannes Zschocke, and Claus Peter Schmitt
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0301 basic medicine ,Dipeptidase ,medicine.medical_specialty ,Dipeptidases ,Carnosine ,Type 2 diabetes ,Diabetic nephropathy ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Risk Factors ,Internal medicine ,Diabetes mellitus ,Genetics ,medicine ,Carnosinemia ,Animals ,Humans ,Diabetic Nephropathies ,Amino Acid Metabolism, Inborn Errors ,Genetics (clinical) ,Polymorphism, Genetic ,biology ,Brain Diseases, Metabolic, Inborn ,Glomerulonephritis ,Protective Factors ,medicine.disease ,Prognosis ,030104 developmental biology ,Endocrinology ,chemistry ,Diabetes Mellitus, Type 2 ,Mutation ,biology.protein ,030217 neurology & neurosurgery ,Kidney disease - Abstract
Carnosinase (CN1) is a dipeptidase, encoded by the CNDP1 gene, that degrades histidine-containing dipeptides, such as carnosine, anserine and homocarnosine. Loss of CN1 function (also called carnosinase deficiency or aminoacyl-histidine dipeptidase deficiency) has been reported in a small number of patients with highly elevated blood carnosine concentrations, denoted carnosinaemia; it is unclear whether the variety of clinical symptoms in these individuals is causally related to carnosinase deficiency. Reduced CN1 function should increase serum carnosine concentrations but the genetic basis of carnosinaemia has not been formally confirmed to be due to CNDP1 mutations. A CNDP1 polymorphism associated with low CN1 activity correlates with significantly reduced risk for diabetic nephropathy, especially in women with type 2 diabetes, and may slow progression of chronic kidney disease in children with glomerulonephritis. Studies in rodents demonstrate antiproteinuric and vasculoprotective effects of carnosine, the precise molecular mechanisms, however, are still incompletely understood. Thus, carnosinemia due to CN1 deficiency may be a non-disease; in contrast, carnosine may potentially protect against long-term sequelae of reactive metabolites accumulating, e.g. in diabetes and chronic renal failure.
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- 2017
48. Prolidase is a critical enzyme for complete gliadin digestion in Tenebrio molitor larvae
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Valeriia F, Tereshchenkova, Irina A, Goptar, Dmitry P, Zhuzhikov, Mikhail A, Belozersky, Yakov E, Dunaevsky, Brenda, Oppert, Irina Yu, Filippova, and Elena N, Elpidina
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Gastrointestinal Tract ,Dipeptidases ,Larva ,Animals ,Insect Proteins ,Membrane Transport Proteins ,Amino Acid Sequence ,Tenebrio ,Transcriptome ,Gliadin ,Substrate Specificity - Abstract
Prolidase is a proline-specific metallopeptidase that cleaves imidodipeptides with C-terminal Pro residue. Prolidase was purified and characterized from the Tenebrio molitor larval midgut. The enzyme was localized in the soluble fraction of posterior midgut tissues, corresponding to a predicted cytoplasmic localization of prolidase according to the structure of the mRNA transcript. Expression of genes encoding prolidase and the major digestive proline-specific peptidase (PSP)-dipeptidyl peptidase 4-were similar. The pH optimum of T. molitor prolidase was 7.5, and the enzyme was inhibited by Z-Pro, indicating that it belongs to type I prolidases. In mammals, prolidase is particularly important in the catabolism of a proline-rich protein-collagen. We propose that T. molitor larval prolidase is a critical enzyme for the final stages of digestion of dietary proline-rich gliadins, providing hydrolysis of imidodipeptides in the cytoplasm of midgut epithelial cells. We propose that the products of hydrolysis are absorbed from the luminal contents by peptide transporters, which we have annotated in the T. molitor larval gut transcriptome. The origin of prolidase substrates in the insect midgut is discussed in the context of overall success of grain feeding insects.
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- 2017
49. The value of prolidase enzyme in rats with experimentally induced mild and severe pancreatitis
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V. Oter, Abdullah Ozgonul, Metin Yalcin, Yalcin, M., Oter, V, Ozgonul, A., Sakarya Üniversitesi/Tıp Fakültesi/Cerrahi Tıp Bilimleri Bölümü, and Öter, Volkan
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Male ,Economics and Econometrics ,medicine.medical_specialty ,Dipeptidases ,medicine.medical_treatment ,Intraperitoneal injection ,Gastroenterology ,Severity of Illness Index ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,Materials Chemistry ,Media Technology ,medicine ,Animals ,Rats, Wistar ,Saline ,Pancreas ,chemistry.chemical_classification ,business.industry ,030208 emergency & critical care medicine ,Forestry ,Metabolism ,medicine.disease ,Rats ,medicine.anatomical_structure ,Enzyme ,chemistry ,Pancreatitis ,Metabolic control analysis ,Acute Disease ,Amylases ,Acute pancreatitis ,Collagen ,business ,Ceruletide - Abstract
INTRODUCTION Acute pancreatitis is basically considered as activation of inactive proenzymes in the pancreas and digestion of the gland itself. This study was performed to determine if prolidase enzyme, which plays a role in collagen metabolism, could be used as a parameter to assess the severity of pancreatitis in experimentally induced mild and severe pancreatitis. MATERIAL AND METHOD To create experimentally induced acute pancreatitis 0.1 ml of normal saline solution (NSS) was given five times with an interval of one hour to rats in the first group; 50 µg/kg of cerulein five times with an interval of one hour in the second group; 80 µg/kg of cerulein five times with an interval of one hour in the third group, in the form of intraperitoneal injection. RESULTS When the serum prolidase values at beginning, 1st, 5th and 24th hours in group II and III were compared among themselves, there was a statistically significant increase(p < 0.05). The evaluation between groups revealed a statistically significant increase in the value of serum prolidase in group II and group III compared with the control group (p < 0.05). In comparisons performed with tissue values, a statistically significant increase determined in the value of serum prolidase in group II and group III compared with the control group was observed (p < 0.05). CONCLUSION The findings obtained in our study showed that prolidase activity increases directly proportionaly with the severity of pancreatitis. This allows us to postulate that prolidase enzyme activities provide guidance about the metabolism of collagen in patients with acute pancreatitis, serious damage occurring in collagen protein and metabolic control is further distorted depending on the duration and intensity of damage but to be able to speak more precisely, there is a need for further, more detailed and extensive researchs (Tab. 8, Fig. 2, Ref. 30).
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- 2017
50. Carnosine Attenuates the Development of both Type 2 Diabetes and Diabetic Nephropathy in BTBR ob/ob Mice
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Maaike Schilperoort, Bernhard K. Krämer, Emile de Heer, Sibylle J. Hauske, Luca Regazzoni, Giancarlo Aldini, Jana D. Braun, Jiedong Qiu, Hannes Koppel, Alessandra Altomare, Hans J. Baelde, Shiqi Zhang, Angelica Rodriguez, Jacob van den Born, Benito A. Yard, Diego O. Pastene, Thomas Albrecht, Alessandra Denisi, Groningen Kidney Center (GKC), Vascular Ageing Programme (VAP), and Groningen Institute for Organ Transplantation (GIOT)
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0301 basic medicine ,Blood Glucose ,Male ,Dipeptidases ,medicine.medical_treatment ,Kidney Glomerulus ,Carnosine ,Administration, Oral ,Gene Expression ,Mice, Obese ,Podocyte ,ACROLEIN ,GLUCOSE ,Diabetic nephropathy ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,CARBONYL STRESS ,Insulin ,Diabetic Nephropathies ,INSULIN-RESISTANCE ,Multidisciplinary ,C-Peptide ,Organ Size ,Glomerular Mesangium ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,medicine.symptom ,medicine.medical_specialty ,METABOLISM ,Article ,Nephropathy ,RATS ,03 medical and health sciences ,Insulin resistance ,Internal medicine ,Diabetes mellitus ,medicine ,Albuminuria ,Animals ,Humans ,Hypoglycemic Agents ,Glycated Hemoglobin ,business.industry ,IN-VITRO ,MASS-SPECTROMETRY ,medicine.disease ,Disease Models, Animal ,RENAL-DISEASE ,030104 developmental biology ,Endocrinology ,chemistry ,Diabetes Mellitus, Type 2 ,CELL LINE ,business - Abstract
We previously demonstrated that polymorphisms in the carnosinase-1 gene (CNDP1) determine the risk of nephropathy in type 2 diabetic patients. Carnosine, the substrate of the enzyme encoded by this gene, is considered renoprotective and could possibly be used to treat diabetic nephropathy (DN). In this study, we examined the effect of carnosine treatment in vivo in BTBR (Black and Tan, BRachyuric) ob/ob mice, a type 2 diabetes model which develops a phenotype that closely resembles advanced human DN. Treatment of BTBR ob/ob mice with 4 mM carnosine for 18 weeks reduced plasma glucose and HbA1c, concomitant with elevated insulin and C-peptide levels. Also, albuminuria and kidney weights were reduced in carnosine-treated mice, which showed less glomerular hypertrophy due to a decrease in the surface area of Bowman’s capsule and space. Carnosine treatment restored the glomerular ultrastructure without affecting podocyte number, resulted in a modified molecular composition of the expanded mesangial matrix and led to the formation of carnosine-acrolein adducts. Our results demonstrate that treatment with carnosine improves glucose metabolism, albuminuria and pathology in BTBR ob/ob mice. Hence, carnosine could be a novel therapeutic strategy to treat patients with DN and/or be used to prevent DN in patients with diabetes.
- Published
- 2017
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