Protective Effect Of Tacrolimus On Podocytes Injured By Puromycin

Objective To observe the effect of tacrolimus on podocyte injury induced by purinomycin,and to explore the mechanism of tacrolimus in protecting podocytes by regulating podocyte autophagy.MethodsMouse glomerular podocytes were cultured in vitro.Puromycin acted on podocytes to establish podocyte injury model.The experiment were divided into control group,solvent group,model group and FK506 group.After 12,24 and 48 hours of treatment,the morphology of podocytes was observed under microscope,the viability of podocytes were tested by cell counting kit-8(CCK-8)assays,and the expression of LC3,?-actin and Nephrin protein were detected by Western blot.Results1.Cell morphology and number changes: At 12 h,24h and 48 h,compared with the control group,the cell area and number of the drug solvent group were basically the same,and there was no significant difference.The area and number of cells in the model group decreased at each time point compared with the control group.The area and number of cells in the experimental groupwere higher than those in the model group at each time point.2.Changes in cell viability: There was no significant difference in podocyte viability between drug solvent group and control group(P>0.05).Compared with the control group,the podocyte viability in the model group decreased significantly(P<0.05).The podocyte viability of the experimental group were significantly higher than that of the model group(P<0.05).3.Protein expression changes: 48 hours later,all histones were collected.The expression levels of LC3 and Nephrin in the drug solvent group were basically the same as those in the control group,with no significant difference.The expression of Nephrin protein in model group was lower than that in control group,and the ratio of LC3II/LC3 I was not increased.Compared with the model group,the expression of Nephrin protein increased and the ratio of LC3II/LC3 I increased in the experimental group.ConclusionsFK506 can up-regulate podocyte autophagy,increase Nephrin protein expression and improve podocyte injury induced by purinomycin.