title: Down-Regulation of MicroRNA-137 Improves High Glucose-Induced Oxidative Stress Injury in Human Umbilical Vein Endothelial Cells by Up-Regulation of AMPKα1.
abstract: OBJECTIVE
  To investigate the effects of miR-137 on high glucose (HG)-induced vascular injury, and to establish the mechanism underlying these effects.
  
  
  METHODS
  Human umbilical vein endothelial cells (HUVECs) were transfected with miR-137 inhibitor or mimic, and then treated with normal or high glucose. Cell viability and apoptosis were detected by using the Cell Counting Kit-8 (CCK-8) assay and flow cytometry, respectively. Reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD) were detected by fluorescent probe (DCFH-DA), thiobarbituric acid reaction, and the nitroblue tetrazolium assay, respectively. The mRNA and protein expressions of AMPKα1 were determined by qRT-PCR and Western blotting.
  
  
  RESULTS
  Down-regulation of miR-137 dramatically reverted HG-induced decreases in cell viability and SOD levels and increases in apoptosis, ROS and MDA levels. Moreover, bioinformatics analysis predicted that the AMPKα1 was a potential target gene of miR-137. Luciferase reporter assay demonstrated that miR-137 could directly target AMPKα1. AMPKα1 overexpression had the similar effect as miR-137 inhibition. Down-regulation of AMPKα1 in HUVECs transfected with miR-137 inhibitor partially reversed the protective effect of miR-137 inhibition on HG-induced oxidative stress in HUVECs.
  
  
  CONCLUSIONS
  Down-regulation of miR-137 ameliorates HG-induced injury in HUVECs by overexpression of AMPKα1, leading to increasing cellular reductive reactions and decreasing oxidative stress. These results provide further evidence for protective effect of miR-137 inhibition on HG-induced vascular injury.
authors: Department of Anesthesiology, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China.
authors: Jie
authors: J
authors: Li
authors: Junfeng
authors: J
authors: Li
authors: Tingting
authors: T
authors: Wei
authors: Junhua
authors: J
authors: Li
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000235:genetics* / Q000378:metabolism; D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D002470:Cell Survival; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005947:Glucose / Q000633:toxicity*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008315:Malondialdehyde / Q000378:metabolism; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D009843:Oligoribonucleotides / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: C545567:MIRN137 microRNA, human; D035683:MicroRNAs; D009843:Oligoribonucleotides; D017382:Reactive Oxygen Species; D008315:Malondialdehyde; D008156:Luciferases; D013482:Superoxide Dismutase; D055372:AMP-Activated Protein Kinases; C526180:PRKAA1 protein, human; D005947:Glucose