Mechanism Of Anemoside B4 In The Intervention Of Focal Segmental Glomerulosclerosis In Rats Based On Podocyte P53 Apoptosis Pathway

Objective: to investigate the effect and mechanism of Anemoside B4 on podocyte apoptosis by establishing a model of doxorubicin induced podocyte apoptosis in vitro,and establish a model of doxorubicin-induced focal segmental-glomerular sclerosis(FSGS)in rats,and to observe the therapeutic effect of Anemoside B4 on FSGS,so as to determine whether the treatment of FSGS by Anemoside B4 is related to the anti-podocytosis of Anemoside B4.Method:1.The podocyte apoptosis model was established by Adriamycin,the activity of podocyte was detected by CCK-8,and the expression of Nephrin and podocin were detected by fluorescence quantitative PCR The expressions of P53,Bcl-2,Bax and CASPASE-3 MRNA and protein in podocytes were detected by fluorescence quantitative PCR and Western Blot.2.The model of focal segmental glomerulosclerosis was established by unilateral nephrectomy combined with tail vein injection of 2.5 mg / kg doxorubicin.After 2weeks,the Serum Bun,Cre,TP,ALB,TCHOL,TG,urinary protein were measured every week,and the pathological changes of kidney were detected by PAS staining,Masson staining was used to detect the degree of renal fibrosis and Glomerulosclerosis,and the ultrastructure of podocyte was examined by transmission electron microscope.The expression of podocyte-related proteins nephrin and podocin in renal tissue and urine,and the expression of P53,Bcl-2,Bax and Caspase-3 m RNA and protein were detected by Western Blot.Conclusion:1.Anemoside B4 has no toxicity to normal MPC-5 cells.The protective effect of P53,Caspase-3 and Bcl-2 / Bax on MPC-5 cells may be related to down-regulation of P53,Caspase-3 and up-regulation of the ratio of Bcl-2 / Bax.2.Anemoside B4 has therapeutic effect on ADR induced FSGS.The renal function could be improved by decreasing the contents of Bun,Cre,TCHOL,TG and urinary protein,and increasing the contents of TP and Alb.The mechanism of the protection of FSGS might be related to the decrease of P53,Caspase-3 and the increase of Bcl-2/ Bax ratio in podocyte,the improvement of Podocyte ultrastructure and the reduction of Renal Intrinsic Cell Apoptosis are related.