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
| 作者: |
1潘雪,
2高玉华,
1韩霞,
3刘云惠
1 内蒙古医科大学附属医院肾内科，呼和浩特 010050 2 中国人民解放军总医院消毒供应室，北京 100036 3 内蒙古医科大学附属医院消化科，呼和浩特 010050 |
| 通讯: |
刘云惠
Email: 1209052554@qq.com |
| DOI: | 10.3978/j.issn.2095-6959. 2018.12.003 |
| 基金: | 解放军总后勤部卫生部资助项目(BWS15B023)。 |
摘要
目的：探讨抑制HPIP基因表达对TGF - β 1诱导的肾小管上皮细胞上皮-间充质转化(ep ithel ial-mesenchymal transition，EMT)及凋亡的影响。方法：10 ng/mL的TGF-β1刺激HK-2细胞24，48 h， 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-cadher in，α-SMA，N-cadher in，Snai l，Tw ist， t-AKT，p-AKT和Bax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， 48 h后HPIP蛋白表达均显著高于对照组(0 h) (P<0.05)。TGF-β1组E-cadherin蛋白表达显著低于空白组，α-SMA，N-cadherin，Snail，Tw ist，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， Snai l，Tw i st，p-AKT和Ba x蛋白表达及细胞凋亡率均显著低于TGF-β1组(P<0.05)。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。
关键词：
肾小管上皮细胞；HPIP基因；TGF-β1诱导；上皮-间充质转化；凋亡
Effect of HPIP gene siRNA on epithelial-mesenchymal transition and apoptosis induced by TGF-β1 in renal tubular epithelial cells
CorrespondingAuthor: LIU Yunhui Email: 1209052554@qq.com
DOI: 10.3978/j.issn.2095-6959. 2018.12.003
Foundation: The work was supported by the Project of Health Subdepartment of PLA General Foundation Logistics Department, China (BWS15B023).
Abstract
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.
Keywords：
renal tubular epithelial cells; HPIP gene; TGF-β1 induction; epithelial-mesenchymal transition; apoptosis