Regulation of Epithelial-Mesenchymal Transition of Glomerular Podocytes by Suoquan Yishen(缩泉益肾)Formula Containing Serum through Inhibition of KDM6A
Objective:To investigate the mechanism by which Suoquan Yishen(缩泉益肾)Formula containing serum improves epithelial-mesen-chymal transition(EMT)and injury of podocytes by inhibiting the expression of histone demethylase KDM6A and regulating EMT-related markers and podocyte injury factors.Methods:Except for the normal control group,after podocyte injury by high glucose stress,the experiment was randomly divided into model control group,14.4 g/kg Suoquan Yishen Formula containing serum of 5％,10％,and 20％groups,and 4.45 μg/mL Cagliflozin group.The morphological changes,viability,mobility,and apoptosis rate of the podocyte were detected by podocyte morphology assay,CCK-8 method,cell scratch assay,and flow cytometry,respectively.RT-qPCR and Western blot were used to detect the mR-NA and protein expressions of Kdm6a,Krüppel-like factor 10(Klf10),podocyte lytic membrane protein(Nephrin),α-smooth muscle actin(α-Sma),N-cadherin(N-cadherin),and P-cadherin(P-cadherin).Results:Compared with the normal control group,the model control group significantly decreased podocyte viability and mobility and increased apoptosis rate(P＜0.01).The mRNA and protein expressions of Kdm6a,Klf10,α-Sma and N-cadherin were significantly up-regulated(P＜0.01),while the mRNA and protein expressions of P-cadherin and Nephrin were significantly down-regulated(P＜0.01).Compared with the model control group,podocyte viability and mobility were significant-ly increased(P＜0.01),and apoptosis rate was significantly decreased(P＜0.01)in the 14.4 g/kg Suoquan Yishen Formula containing ser-um of 5％and 10％groups and 4.45 μg/mL Cagliflozin group.In the 14.4 g/kg Suoquan Yishen Formula containing serum of 10％group and 4.45 μg/mL Cagliflozin group,the mRNA and protein expressions of Kdm6a,Klf10,α-Sma and N-cadherin were down-regulated(P＜0.01 or P＞0.05),while those of P-cadherin and Nephrin were significantly up-regulated(P＜0.05 or P＜0.01).Conclusion:Suoquan Yish-en Formula containing serum may improve EMT and injury of podocytes by inhibiting KDM6A and regulating EMT-related markers and podo-cyte injury factors.
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