Study on the mechanism of Shenyan Fangshuai Formula on induction mouse kidney podocytes injury and autophagy by phenyl sulfate
[Objective]To observe the effect of Shenyan Fangshuai Formula on phenyl sulfate(PS)-induced mouse kidney podocyte injury and autophagy-related protein expression,and to explore its intervention effect on podocyte injury and autophagy.[Methods]We selected SD rats and gavaged Shenyan Fangshuai Formula and valsartan aqueous solution to prepare low,medium and high concentration drug-containing serum and western control serum,respectively.Podocytes were cultured in vitro and divided into normal control(Con)group,PS stimulation(PS)group,Shenyan Fangshuai Formula low,medium,and high three concentration treatment groups(SYFS-L,SYFS-M,and SYFS-H),and valsartan treatment(XST)group.The CCK-8 method screened the most appropriate intervention concentration and administration concentration of medicated serum.qRT-PCR and Western blot were used to detect Podocin,CD2AP,BNIP3,p62,and LC3Ⅱ/Ⅰ mRNA and protein expression.[Results]CCK-8 method to detect the cellular value-added rate identified 190 μmol/L concentration of PS to treat the podocytes for 24h as the modeling method for this experiment.Compared with the Con group,CD2AP,Podocin,BNIP3,LC3Ⅱ/Ⅰprotein and mRNA expression were significantly down-regulated and p62 protein and mRNA expression were significantly up-regulated in the PS group.Compared with the PS group,the Shenyan Fangshuai Formula group could significantly promote the expression of CD2AP,Podocin,BNIP3,LC3 mRNA and protein,and reduce the expression of p62,and it was better than that in the valsartan group.[Conclusion]Phenyl sulfate may cause podocyte injury by inhibiting podocyte mitochondrial autophagy,while Shenyan Fangshuai Formula ameliorates podocyte injury by up-regulating LC3,BNIP3,and down-regulating p62 protein expression,enhancing cellular autophagy,and inhibiting apoptosis.
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