title: Ramipril protects the endothelium from high glucose-induced dysfunction through CaMKKβ/AMPK and heme oxygenase-1 activation.
abstract: This study aims to investigate the effects of ramipril (RPL) on endothelial dysfunction associated with diabetes mellitus using cultured human aortic endothelial cells (HAECs) and a type 2 diabetic animal model. The effect of RPL on vasodilatory function in fat-fed, streptozotocin-treated rats was assessed. RPL treatment of 8 weeks alleviated insulin resistance and inhibited the decrease in endothelium-dependent vasodilation in diabetic rats. RPL treatment also reduced serum advanced glycation end products (AGE) concentration and rat aorta reactive oxygen species formation and increased aorta endothelium heme oxygenase-1 (HO-1) expression. Exposure of HAECs to high concentrations of glucose induced prolonged oxidative stress, apoptosis, and accumulation of AGEs. These effects were abolished by incubation of ramiprilat (RPT), the active metabolite of RPL. However, treatment of HAECs with STO-609, a CaMKKβ (Ca(2+)/calmodulin-dependent protein kinase kinase-β) inhibitor; compound C, an AMPK (AMP-activated protein kinase) inhibitor; and Zn(II)PPIX, a selective HO-1 inhibitor, blocked these beneficial effects of RPT. In addition, RPT increased nuclear factor erythroid 2-related factor-2 (Nrf-2) nuclear translocation and activation in a CaMKKβ/AMPK pathway-dependent manner, leading to increased expression of the Nrf-2-regulated antioxidant enzyme, HO-1. The inhibition of CaMKKβ or AMPK by pharmaceutical approach ablated RPT-induced HO-1 expression. Taken together, RPL ameliorates insulin resistance and endothelial dysfunction in diabetes via reducing oxidative stress. These effects are mediated by RPL activation of CaMKK-β, which in turn activates the AMPK-Nrf-2-HO-1 pathway for enhanced endothelial function.
authors.affiliation: Key Laboratory of Exercise and Health Sciences of Ministry of Education, Shanghai University of Sport, Shanghai, China (S.T., Y.L.); Department of Biochemistry, University of California, Riverside, California (S.T.); Shandong Sport University, Shandong, China (X.G.); Center for Animal Experiment/ABSL-3 Laboratory, Wuhan University, Hubei, China (K.W.); Center for Medical Research, Wuhan University, Hubei, China (K.W.); and Department of Basic Theories, Hubei University of Traditional Chinese Medicine, Hubei, China (H.Y.).
authors.forename: Shiliu
authors.initials: S
authors.lastname: Tian
authors.affiliation: Key Laboratory of Exercise and Health Sciences of Ministry of Education, Shanghai University of Sport, Shanghai, China (S.T., Y.L.); Department of Biochemistry, University of California, Riverside, California (S.T.); Shandong Sport University, Shandong, China (X.G.); Center for Animal Experiment/ABSL-3 Laboratory, Wuhan University, Hubei, China (K.W.); Center for Medical Research, Wuhan University, Hubei, China (K.W.); and Department of Basic Theories, Hubei University of Traditional Chinese Medicine, Hubei, China (H.Y.).
authors.forename: Xinfa
authors.initials: X
authors.lastname: Ge
authors.affiliation: Key Laboratory of Exercise and Health Sciences of Ministry of Education, Shanghai University of Sport, Shanghai, China (S.T., Y.L.); Department of Biochemistry, University of California, Riverside, California (S.T.); Shandong Sport University, Shandong, China (X.G.); Center for Animal Experiment/ABSL-3 Laboratory, Wuhan University, Hubei, China (K.W.); Center for Medical Research, Wuhan University, Hubei, China (K.W.); and Department of Basic Theories, Hubei University of Traditional Chinese Medicine, Hubei, China (H.Y.).
authors.forename: Ke
authors.initials: K
authors.lastname: Wu
authors.affiliation: Key Laboratory of Exercise and Health Sciences of Ministry of Education, Shanghai University of Sport, Shanghai, China (S.T., Y.L.); Department of Biochemistry, University of California, Riverside, California (S.T.); Shandong Sport University, Shandong, China (X.G.); Center for Animal Experiment/ABSL-3 Laboratory, Wuhan University, Hubei, China (K.W.); Center for Medical Research, Wuhan University, Hubei, China (K.W.); and Department of Basic Theories, Hubei University of Traditional Chinese Medicine, Hubei, China (H.Y.).
authors.forename: Huabing
authors.initials: H
authors.lastname: Yang
authors.affiliation: Key Laboratory of Exercise and Health Sciences of Ministry of Education, Shanghai University of Sport, Shanghai, China (S.T., Y.L.); Department of Biochemistry, University of California, Riverside, California (S.T.); Shandong Sport University, Shandong, China (X.G.); Center for Animal Experiment/ABSL-3 Laboratory, Wuhan University, Hubei, China (K.W.); Center for Medical Research, Wuhan University, Hubei, China (K.W.); and Department of Basic Theories, Hubei University of Traditional Chinese Medicine, Hubei, China (H.Y.) yuliu2006@126.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism*; D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000502:physiology; D017209:Apoptosis / Q000187:drug effects; D001562:Benzimidazoles / Q000494:pharmacology; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002478:Cells, Cultured; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000139:chemically induced / Q000188:drug therapy* / Q000378:metabolism / Q000503:physiopathology; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D004789:Enzyme Activation / Q000187:drug effects; D005947:Glucose / Q000378:metabolism*; D017127:Glycation End Products, Advanced / Q000097:blood; D006419:Heme Oxygenase (Decyclizing) / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D007333:Insulin Resistance / Q000502:physiology; D008297:Male; D051267:NF-E2-Related Factor 2 / Q000378:metabolism; D053644:Naphthalimides / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011524:Protoporphyrins / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D017257:Ramipril / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014664:Vasodilation / Q000187:drug effects / Q000502:physiology
chemical_list: D001562:Benzimidazoles; D017127:Glycation End Products, Advanced; D051267:NF-E2-Related Factor 2; D053644:Naphthalimides; C495637:Nfe2l2 protein, rat; D047428:Protein Kinase Inhibitors; D011524:Protoporphyrins; D011720:Pyrazoles; D011743:Pyrimidines; D017382:Reactive Oxygen Species; C458525:STO 609; C516138:dorsomorphin; C017803:zinc protoporphyrin; C052549:ramiprilat; D006419:Heme Oxygenase (Decyclizing); C491523:Hmox1 protein, rat; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase; D055372:AMP-Activated Protein Kinases; D005947:Glucose; D017257:Ramipril