MicroRNA-193a regulating Wilms'tumor gene 1 to promote mouse podocyte apoptosis in diabetic nephropathy / 解剖学报

ABSTRACT

Objective To explore the effect of microRNA( miR) -193a on the apoptosis of mouse podocytes in diabetic nephropathy (DN) and its mechanism. Methods The DN model was replicated by culturing podocytes with high glucose in vitro and intraperitoneal injection of streptozocin (STZ) in mice in vivo. The cells or 60 mice were randomly divided into normal control (NC) group, model control group, and miR-NC inhibitor group, miR-193a inhibitor group, miR-NC mimic group and miR-193a mimic group. Flow cytometry, immunohistochemistry, TUNEL, Real-time PCR, Western blotting were used to examine the apoptosis of DN mice and mouse podocytes. Results The expression of Nephrin and Podocin in podocytes was weakened in DN mice and renal podocytes induced by high glucose, the apoptotic rate increased significantly, miR-193a was highly expressed, the levels of cleaved-Caspase-3 and Bax protein increased significantly, the level of Bcl-2 protein decreased significantly, and miR-193a inhibitor could improve this process. Wilms' tumor gene 1 ( WT1 ) mRNA and protein expression levels were significantly reduced in DN mice and podocytes cultured with high glucose. WT1 protein expression increased significantly after miR-193a inhibitor intervention, and WT1 protein expression was significantly reduced after miR-193a mimic transfection. Up-regulating WT1 could reduce the effect of miR- 193a on the apoptosis of mouse podocytes induced by high glucose. The dual luciferase reporter experiment confirmed the targeting relationship between miR-193a and WT1. Conclusion MiR-193a down-regulates the expression of WT1 and promotes apoptosis of DN podocytes.