HPIP基因siRNA对TGF-β1诱导的肾小管上皮细胞上皮-间充质转化及凋亡的影响

Effect of HPIP gene siRNA on epithelial-mesenchymal transition and apoptosis induced by TGF-β1 in renal tubular epithelial cells

目的:探讨抑制HPIP基因表达对TGF-β1诱导的肾小管上皮细胞上皮-间充质转化(epithelial-mesenchymal transition,EMT)及凋亡的影响。方法:10ng/mL的TGF-β1刺激HK-2细胞24,48h,Western印迹法检测HPIP蛋白表达。将HK-2细胞随机分为空白组、TGF-β1组和TGF-β1+si-HPIP组,处理24 h后,Western印迹法检测HPIP,E-cadherin,α-SMA,N-cadherin,Snail,Twist,t-AKT,p-AKT和Ba x蛋白表达;流式细胞术检测细胞凋亡率。结果:TGF-β1刺激HK-2细胞24,48h后HPIP蛋白表达均显著高于对照组(0h)(P<0.05)。TGF-β1组E-cadherin蛋白表达显著低于空白组,α-SMA,N-cadherin,Snail,Twist,p-AKT和Bax蛋白表达及细胞凋亡率均显著高于空白组(P<0.05),而TGF-β1+si-HPIP组E-cadherin蛋白表达显著高于TGF-β1组,α-SMA,N-cadherin,Snail, Tw i st, p-A KT和Ba x蛋白表达及细胞凋亡率均显著低于TG F-β1组(P <0. 0 5)。结论:抑制HPIP基因表达可延缓肾小管上皮细胞EMT进程和降低细胞凋亡率,机制可能与下调PI3K/AKT信号有关。

Objective: To investigate the effect of HPIP gene expression was inhibited on the epithelial-mesenchymal transition (EMT) and apoptosis of renal tubular epithelial cells induced by TGF-β1. Methods: After HK-2 cells were stimulated with 10 ng/mL TGF-β1 for 24 h and 48 h, Western blot was used to detect the expression of HPIP protein. HK-2 cells were randomly divided into a blank group, a TGF-β1 group and a TGF-β1 + si-HPIP group. The cells were treated with 24 h, Western blot were used to detect HPIP, E-cadherin,α-SMA, N-cadherin, Snail, Twist, t-AKT, p-AKT and Bax protein expression. The apoptosis rate of cells was detected by flow cytometry. Results: The expression of HPIP protein in HK-2 cells stimulated by TGF-β1 for 24 h and 48 h was significantly higher than that in the control group (0 h)(P<0.05). The expression of E-cadherin protein in TGF-β1 group was significantly lower than that in the blank group, the expression of α-SMA, N-cadherin, Snail, Twist, p-AKT, Bax protein and cells apoptosis were significantly higher than those in the blank group (P<0.05), but the expression of E-cadherin protein in TGF-β1+si-HPIP group was significantly higher than those in TGF-β1 group, the expression of α-SMA, N-cadherin, Snail, Twist, p-AKT protein and cells apoptosis were significantly lower than those in TGF-β1 group (P<0.05). Conclusion: Inhibition of HPIP gene expression can reduce the EMT process and apoptosis rate of renal tubular epithelial cells, which may be related to downregulation of PI3K/AKT signaling.