Down Regulation of MicroRNA 137 Improves High Glucose Induced Oxidative Stress Injury in Human Umbilical Vein Endothelial Cells by Up Regulation of AMPKa1
Background/Aims: 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 AMPKa1 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 AMPKa1 was a potential target gene of miR 137. Luciferase reporter assay demonstrated that miR 137 could directly target AMPKa1. AMPKa1 overexpression had the similar effect as miR 137 inhibition. Down regulation of AMPKa1 in HUVECs transfected with miR 137 inhibitor partially reversed the protective effect of miR 137 inhibition on HG induced oxidative stress in HUVECs. Conclusion: Down regulation of miR 137 ameliorates HG induced injury in HUVECs by overexpression of AMPKa1, 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.
Cellular Physiology and Biochemistry