Overexpression of MIR-302C-3P Leads to Apoptosis in Human Renal Mesangial Cells

Objective: The apoptosis of glomerular mesangial cells is closely related to the occurrence and development of diabetic nephropathy (DN). However, the mechanism remains unclear. In the present study, we found that the high-glucose cultures induced the upregulation of the miRNA-302c-3p. This study aimed to investigate the effects of over expression of miR-302c-3p on human glomerular mesangial cells (HRMCs). Materials and Methods: HRMCs were cultured in vitro and divided into 2 groups: ①: siRNA-NC group, ② siRNA-TIMP3 group. The miR-302c-3p expression was up-regulated in HRMCs by using miR-302c-3p mimic. We measured the expression of MDA, SOD, CAT, and ROS to examine the oxidative stress of cells. The apoptosis rate of cells was determined by flow cytometry, and the mitochondrial membrane potential (MMP) was examined by the JC-1 detection kit. Real-time polymerase chain reaction (PCR) was used to detect the relative expression of p53 and Survivin. Protein expression of Bcl-2, BAX, caspase-3, p27, Apaf-1, Cytochrome C, and β-actin was analyzed by Western blot. Results: Compared with the control group, we found that the overexpression of miR-302c-3p significantly increased the oxidative stress and apoptosis on HRMCs. The level of p53 was significantly increased ( p <0.01), and the level of Survivin was reduced significantly ( p <0.05) in the miR-302c-3p mimic group. Western blot results showed that the expression of BAX, p27, Apaf-1 and Cytochrome C was significantly up-regulated; the expression of Bcl-2 was significantly down-regulated in the HRMCs in miR-302c-3p mimic group. Conclusions: These findings indicated that overexpression of miR-302c-3p induced the apoptosis of HRMCs, and might be associated with the oxidative stress.