Effect of Smad4 Lentivirus Transfection on Vancomycin-induced EMT in Renal Tubular Epithelial Cells
Objective:To explore the effect of lentivirus-mediated overexpression of Smad4 or silencing of Smad4 on the epithelial-mesenchymal transition(EMT)of renal tubular epithelial cells HK-2 induced by vancomycin.Methods:Design and package Smad4 silenced or overexpressed chronic virus,and then use the obtained lentivirus to transiently transfect HK-2,the HK-2 cells were divided into vancomycin group,empty vector group(transfected with HBLV-GFP-PURO),Smad4 overexpression group(transfected with HBLV-h-Smad4-GFP-PURO),and Smad4 silencing group(transfected with HBLV-h-Smad4-shR-NA-GFP-PURO),select normal HK-2 cells from the control group.Cell migration ability,oxidative stress index(TGF-β1,SOD,GSH,MDA)in supernatant,protein expression levels of Smad4,E-cadherin,FN,Vimentin were compared in the five groups.Results:Compared with the control group,the migration ability of HK-2 cells,the contents of TGF-β1 and MDA in the su-pernatant,the expression levels of Smad4,FN,Vimentin protein in the vancomycin group and empty vector group were higher(P<0.05),while the contents of SOD and GSH,the expression level of E-cadherin protein were lower(P<0.05).Overexpression of Smad4 increased the migration ability and oxidative stress response of HK-2 cells,and promoted EMT(P<0.05).Silencing Smad4 decreased the migration ability and oxidative stress response of HK-2 cell,and inhibit EMT(P<0.05).Conclusion:Smad4 regu-lates vancomycin-induced oxidative stress and EMT in HK-2 cells.
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