Influence Of Angiotensin-(1-7) On Hypoxic Induced Rat’s Tubular Epithelial Myofibroblast Transdifferentiation

objective To explore the influence of angiotensin-(1-7)[ang-(1-7)] on angiotensionⅡII induced rat’s tubular epithelial myofibroblast transdifferentiation and the secretion of extracellular matrix. Methods The NRK52E were maintained and sub-cultured. Experiments had been performed after the cells were prepared2X105/ml suspension with DMEM and cultured in6-well plates. The cells were divided into four groups according to the different intervention factors, final concentration of Ang-(1-7) and was10-6mol/L Co was100umol/L.①control group:the NRK52E were cultured without any intervention factor;②Co group:the NRK52E were stimulated by Co;③Ang-(1-7) group:the NRK52E were stimulated by Ang-(1-7);④Co+Ang-(1-7) group:the NRK52E were co-stimulated by Co and Ang-(1-7). The NRK52E cells, cultured for1d,3d,6d.(1) we detected the expressions of α-SMA、HIF-1α and p-ERK1/2by immunocytochemistry method.(2) The content of Col I in the cultured supernatant were measured by ELISA.(3) The expressions of p-ERK1/2were detected by Western blot analyses. Results (1) immunocytochemistry:①the change of expressions of α-SMA of tubular epithelial cells:The cells, cultured for1d,3d,6d, expressions of a-SMA of control group were no significant change, Ang-(1-7) group was similar with it, Co group increased from Id to3d than that of control group at the same time (P>0.05), cultured for6d, Co group increased significantly than that of control group (P<0.05), Co+Ang-(1-7) group decreased than that of Co group at the same time (P<0.05);②the change of expressions of HIF-la of tubular epithelial cells:The cells, cultured for6d, expressions of HIF-la of control group were no significant change, Ang-(1-7) group was similar with it, Co group increased significantly than that of control group (P<0.05), Co+Ang-(1-7) group decreased than that of Co group at the same time (P<0.05).③the change of expressions of p-ERK1/2of tubular epithelial cells:The cells, cultured for6d, expressions of p-ERK1/2of control group were no significant change, Ang-(1-7) group was similar with it, Co group increased significantly than that of control group (P<0.05), Co+Ang-(1-7) group decreased than that of Co group at the same time (P<0.05).(2) ELISA:cultured for6d, content of Col I in the Ang-(1-7) group and control group were no significant change, Co group was more obviously advanced than that of control group (P<0.05) and Co+Ang-(1-7) group significantly decreased than that of Co group (P<0.05).(3) Western blot analyses:The cells, cultured for6d, the expression of p-ERK1/2of Ang-(1-7) group were not any changes than that of control group, Co group enhanced the expression of p-ERK1/2 (P<0.05), Co+Ang-(1-7) group significantly inhibited the expression of p-ERK1/2(P<0.05) than that of Co group. conclusion Hypoxia can induce tubular epithelia-myofibroblast transdifferentiation (TEMT). Ang-(1-7) improves Co-induced hypoxie status and inhibits TEMT which might be mediated by the p-ERK1/2signal-transduction pathway.