title: Metformin inhibits vascular calcification in female rat aortic smooth muscle cells via the AMPK-eNOS-NO pathway.
abstract: Metformin exhibits diverse protective effects against diabetic complications, such as bone loss. Here, we investigated the effect of metformin on vascular calcification, another type 2 diabetes complication. In female rat aortic smooth muscle cells (RASMCs), we observed that metformin significantly alleviated β-glycerophosphate-induced Ca deposition and alkaline phosphatase activity, corresponding with reduced expression of some specific genes in osteoblast-like cells, including Runx2 and bone morphogenetic protein-2, and positive effects on α-actin expression, a specific marker of smooth muscle cells. Mechanistic analysis showed that phosphorylation levels of both AMP-activated protein kinase (AMPK) and endothelial nitric oxide synthase (eNOS) were increased with NO overproduction. After inhibition of either AMPK or eNOS with the pharmacologic inhibitors, compound C or Nω-Nitro-L-arginine methyl ester, NO production was lowered and metformin-meditated vascular protection against β-glycerophosphate-induced Ca deposition was removed. Our results support that metformin prevents vascular calcification via AMPK-eNOS-NO pathway.
authors.affiliation: PhD, State Key Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, Fourth Military Medical University, 710032 Xi'an, China, luzfliuq@fmmu.edu.cn; or Qingshen Zhu, MD, PhD, Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, 710032 Xi'an, China, E-mail: zhuqsh@fmmu.edu.cn.
authors.forename: Xiaorui
authors.initials: X
authors.lastname: Cao
authors.forename: Huan
authors.initials: H
authors.lastname: Li
authors.forename: Huiren
authors.initials: H
authors.lastname: Tao
authors.forename: Ning
authors.initials: N
authors.lastname: Wu
authors.forename: Lifeng
authors.initials: L
authors.lastname: Yu
authors.forename: Dawei
authors.initials: D
authors.lastname: Zhang
authors.forename: Xiaozhao
authors.initials: X
authors.lastname: Lu
authors.forename: Jinyu
authors.initials: J
authors.lastname: Zhu
authors.forename: Zifan
authors.initials: Z
authors.lastname: Lu
authors.forename: Qingsheng
authors.initials: Q
authors.lastname: Zhu
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001011:Aorta / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D054338:Cell Transdifferentiation / Q000187:drug effects; D002478:Cells, Cultured; D003925:Diabetic Angiopathies / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000009:adverse effects; D005260:Female; D005994:Glycerophosphates / Q000009:adverse effects / Q000037:antagonists & inhibitors; D007004:Hypoglycemic Agents / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D008687:Metformin / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000009:adverse effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects; D061205:Vascular Calcification / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*
chemical_list: D004791:Enzyme Inhibitors; D005994:Glycerophosphates; D007004:Hypoglycemic Agents; D047428:Protein Kinase Inhibitors; D009569:Nitric Oxide; D008687:Metformin; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse; D055372:AMP-Activated Protein Kinases; C031463:beta-glycerophosphoric acid