Research on Mechanism of Yitangkang(益糖康)Improving Early Renal Injury Induced by Hyperglycemia Based on NFAT5 Mediated EMT
Objective To investigate the effect of nuclear factor of activated T cells 5(NFAT5)on epithelial-mesenchymal transition(EMT)of human renal tubular epithelial HK-2 cells induced by high glucose and the mechanism of Yitangkang(益糖康)improving early renal injury induced by hyperglycemia.Methods Twenty male SD rats were randomly divided into normal sa-line group and Yitangkang group,with 10 rats in each group.Normal saline group was given normal saline by intragastric adminis-tration(1 mL/100 g)and Yitangkang group was given Yitangkang decoction by intragastric administration(2.10 g/100 g),for5 consecutive days.The abdominal aorta blood was taken out and the supernatant was separated by centrifugation.HK-2 cells were induced by high glucose to construct an in vitro model of early diabetic nephropathy and the corresponding drugs were added to intervene the cells.The qRT-PCR and Western Blot assays were performed to detect the expression levels of NFAT5,E-cadherin and Vimentin mRNA and protein in the cells transfected with NC-siRNA(NC-siRNA group)/NFAT5-siRNA(NFAT5-siRNA group)or the cells treated with normal saline drug-containing serum(NS group)/Yitangkang drug-contai-ning serum(YTK group)or the cells treated with Yitangkang drug-containing serum and transfected with GV492-Empty(YTK+GV492-Empty group)/treated with Yitangkang drug-containing serum and transfected with GV492-NFAT5(YTK+GV492-NFAT5 group),respectively.The wound healing and Transwell assay were used to test the migration and invasion abili-ties of cells in different groups.Results Transfection of NFAT5-siRNA could significantly inhibit the expressions of NFAT5 mR-NA and protein(P＜0.01,P＜0.05).Compared with those of the NC-siRNA group,the expression levels of E-cadherin mR-NA and protein(P＜0.05,P＜0.01)in NFAT5-siRNA group were significantly increased,the expression levels of Vimentin mRNA and protein(P＜0.05,P＜0.01)were significantly decreased.The migration and invasion abilities of cells were signifi-cantly decreased(P＜0.05,P＜0.01).Compared with those of the NS group,the expression levels of NFAT5 mRNA and protein in YTK group were significantly decreased(P＜0.05,P＜0.05).Transfection of GV492-NFAT5 could significantly promote the expressions of NFAT5 mRNA and protein(P＜0.05,P＜0.01).Compared with those of the YTK+GV492-Empty group,the expression levels of E-cadherin mRNA and protein in YTK+GV492-NFAT5 group were significantly decreased(P＜0.01,P＜0.01).The expression levels of Vimentin mRNA and protein were significantly increased(P＜0.05,P＜0.01),and the mi-gration and invasion abilities were significantly enhanced(P＜0.05,P＜0.05).Conclusion In high glucose condition,NFAT5 can promote EMT of human renal tubular epithelial cells.Yitangkang can inhibit EMT of human renal tubular epithelial cells by down-regulating NFAT5 expression to improve early renal injury caused by hyperglycemia.
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