PMID- 25352635 OWN - NLM STAT- MEDLINE DCOM- 20150422 LR - 20181113 IS - 1939-327X (Electronic) IS - 0012-1797 (Print) IS - 0012-1797 (Linking) VI - 64 IP - 3 DP - 2015 Mar TI - Hyperhomocysteinemia and hyperglycemia induce and potentiate endothelial dysfunction via mu-calpain activation. PG - 947-59 LID - 10.2337/db14-0784 [doi] AB - Plasma homocysteine (Hcy) levels are positively correlated with cardiovascular mortality in diabetes. However, the joint effect of hyperhomocysteinemia (HHcy) and hyperglycemia (HG) on endothelial dysfunction (ED) and the underlying mechanisms have not been studied. Mild (22 micromol/L) and moderate (88 micromol/L) HHcy were induced in cystathionine beta-synthase wild-type (Cbs(+/+)) and heterozygous-deficient (Cbs(-/+)) mice by a high-methionine (HM) diet. HG was induced by consecutive injection of streptozotocin. We found that HG worsened HHcy and elevated Hcy levels to 53 and 173 micromol/L in Cbs(+/+) and Cbs(-/+) mice fed an HM diet, respectively. Both mild and moderate HHcy aggravated HG-impaired endothelium-dependent vascular relaxation to acetylcholine, which was completely abolished by endothelial nitric oxide synthase (eNOS) inhibitor N(G)-nitro-L-arginine methyl ester. HHcy potentiated HG-induced calpain activation in aortic endothelial cells isolated from Cbs mice. Calpain inhibitors rescued HHcy- and HHcy/HG-induced ED in vivo and ex vivo. Moderate HHcy- and HG-induced mu-calpain activation was potentiated by a combination of HHcy and HG in the mouse aorta. mu-Calpain small interfering RNA (mu-calpsiRNA) prevented HHcy/HG-induced ED in the mouse aorta and calpain activation in human aortic endothelial cells (HAECs) treated with DL-Hcy (500 micromol/L) and d-glucose (25 mmol) for 48 h. In addition, HHcy accelerated HG-induced superoxide production as determined by dihydroethidium and 3-nitrotyrosin staining and urinary 8-isoprostane/creatinine assay. Antioxidants rescued HHcy/HG-induced ED in mouse aortas and calpain activation in cultured HAECs. Finally, HHcy potentiated HG-suppressed nitric oxide production and eNOS activity in HAECs, which were prevented by calpain inhibitors or mu-calpsiRNA. HHcy aggravated HG-increased phosphorylation of eNOS at threonine 497/495 (eNOS-pThr497/495) in the mouse aorta and HAECs. HHcy/HG-induced eNOS-pThr497/495 was reversed by micro-calpsiRNA and adenoviral transduced dominant negative protein kinase C (PKC)beta2 in HAECs. HHcy and HG induced ED, which was potentiated by the combination of HHcy and HG via mu-calpain/PKCbeta2 activation-induced eNOS-pThr497/495 and eNOS inactivation. CI - (c) 2015 by the American Diabetes Association. Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. FAU - Cheng, Zhongjian AU - Cheng Z AD - Center for Metabolic Disease Research, Temple University School of Medicine, Philadelphia, PA Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA. FAU - Jiang, Xiaohua AU - Jiang X AD - Center for Metabolic Disease Research, Temple University School of Medicine, Philadelphia, PA Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA Center for Cardiovascular Research, Temple University School of Medicine, Philadelphia, PA. FAU - Pansuria, Meghana AU - Pansuria M AD - Center for Metabolic Disease Research, Temple University School of Medicine, Philadelphia, PA Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA. FAU - Fang, Pu AU - Fang P AD - Center for Metabolic Disease Research, Temple University School of Medicine, Philadelphia, PA Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA. FAU - Mai, Jietang AU - Mai J AD - Center for Metabolic Disease Research, Temple University School of Medicine, Philadelphia, PA Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA. FAU - Mallilankaraman, Karthik AU - Mallilankaraman K AD - Center for Translational Medicine, Temple University School of Medicine, Philadelphia, PA. FAU - Gandhirajan, Rajesh Kumar AU - Gandhirajan RK AD - Center for Translational Medicine, Temple University School of Medicine, Philadelphia, PA. FAU - Eguchi, Satoru AU - Eguchi S AD - Center for Cardiovascular Research, Temple University School of Medicine, Philadelphia, PA Department of Physiology, Temple University School of Medicine, Philadelphia, PA. FAU - Scalia, Rosario AU - Scalia R AD - Center for Cardiovascular Research, Temple University School of Medicine, Philadelphia, PA Department of Physiology, Temple University School of Medicine, Philadelphia, PA. FAU - Madesh, Muniswamy AU - Madesh M AD - Center for Translational Medicine, Temple University School of Medicine, Philadelphia, PA Department of Biochemistry, Temple University School of Medicine, Philadelphia, PA. FAU - Yang, Xiaofeng AU - Yang X AD - Center for Metabolic Disease Research, Temple University School of Medicine, Philadelphia, PA Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA Center for Cardiovascular Research, Temple University School of Medicine, Philadelphia, PA Center for Thrombosis Research, Temple University School of Medicine, Philadelphia, PA. FAU - Wang, Hong AU - Wang H AD - Center for Metabolic Disease Research, Temple University School of Medicine, Philadelphia, PA Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA Center for Cardiovascular Research, Temple University School of Medicine, Philadelphia, PA Center for Thrombosis Research, Temple University School of Medicine, Philadelphia, PA hongw@temple.edu. LA - eng GR - R01 HL108910/HL/NHLBI NIH HHS/United States GR - R01 HL077288/HL/NHLBI NIH HHS/United States GR - HL-67033/HL/NHLBI NIH HHS/United States GR - R01 DK096521/DK/NIDDK NIH HHS/United States GR - R01 HL117654/HL/NHLBI NIH HHS/United States GR - HL-108910/HL/NHLBI NIH HHS/United States GR - HL-9445/HL/NHLBI NIH HHS/United States GR - R01 HL082774/HL/NHLBI NIH HHS/United States GR - R01 HL110764/HL/NHLBI NIH HHS/United States GR - HL-110764/HL/NHLBI NIH HHS/United States GR - HL-116917/HL/NHLBI NIH HHS/United States GR - HL-82774/HL/NHLBI NIH HHS/United States GR - F32 HL009445/HL/NHLBI NIH HHS/United States GR - HL-117654/HL/NHLBI NIH HHS/United States GR - HL-77288/HL/NHLBI NIH HHS/United States GR - R01 HL067033/HL/NHLBI NIH HHS/United States GR - R01 HL116917/HL/NHLBI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20141028 PL - United States TA - Diabetes JT - Diabetes JID - 0372763 RN - 0 (Blood Glucose) RN - 11062-77-4 (Superoxides) RN - EC 1.14.13.39 (Nitric Oxide Synthase Type III) RN - EC 3.4.22.- (Calpain) RN - EC 3.4.22.- (mu-calpain) RN - EC 4.2.1.22 (Cystathionine beta-Synthase) RN - Homocysteinemia SB - IM CIN - Diabetes. 2015 Mar;64(3):693-5. PMID: 25713192 MH - Animals MH - Blood Glucose/metabolism MH - Calpain/genetics/*metabolism MH - Cells, Cultured MH - Cystathionine beta-Synthase MH - Endothelial Cells/*metabolism/*pathology MH - Humans MH - Hyperglycemia/*metabolism/*physiopathology MH - Hyperhomocysteinemia/*metabolism/*physiopathology MH - Male MH - Mice MH - Mice, Mutant Strains MH - Nitric Oxide Synthase Type III/metabolism MH - Superoxides/metabolism PMC - PMC4338586 EDAT- 2014/10/30 06:00 MHDA- 2015/04/23 06:00 PMCR- 2016/03/01 CRDT- 2014/10/30 06:00 PHST- 2014/10/30 06:00 [entrez] PHST- 2014/10/30 06:00 [pubmed] PHST- 2015/04/23 06:00 [medline] PHST- 2016/03/01 00:00 [pmc-release] AID - db14-0784 [pii] AID - 0784 [pii] AID - 10.2337/db14-0784 [doi] PST - ppublish SO - Diabetes. 2015 Mar;64(3):947-59. doi: 10.2337/db14-0784. Epub 2014 Oct 28.