摘要
目的探讨磷脂酰肌醇3激酶(PI3K)/Akt通路在转化生长因子β1(TGFβ1)体外诱导的人腹膜间皮细胞(HPMC)上皮-间质细胞转分化(EMT)过程中的作用。方法从人腹膜组织中分离和培养人原代腹膜间皮细胞。观察TGFβ1对人腹膜间皮细胞Akt磷酸化的影响,在不同时间点检测p-Akt的表达水平。采用免疫印迹方法检测应用PI3K特异性抑制剂Wortmannin对TGFβ1诱导后细胞内p-Akt、α-平滑肌肌动蛋白(α-SMA)和E-钙黏附蛋白(E-Cadherin)表达的影响。结果10ng/mLTGFβ1刺激HPMC0.5h后p-Akt水平开始升高,1h后达到顶峰,2h后逐渐减弱。TGFβ1诱导HPMC48h后α-SMA表达显著升高,E-cadherin表达显著下降;同时加用Wortmannin则p-Akt、α-SMA表达明显抑制。结论PI3K/Akt信号系统参与TGFβ1介导的人腹膜间皮细胞EMT过程,提示对其通路的抑制可有效阻止TGFβ1介导的腹膜纤维化。
【Objective】To investigate the role of PI3K/Akt signal pathway in transforming growth factor β1 induced epithelia-mesenchymal transition of human peritoneal mesothelial cell. 【Methods】Primary HPMC was harvested from humane omental tissue and maintained under defined in vitro conditions. The protein expression of pAkt was detected at different time point after 10 ng/ml TGFβ1 stimulating. We also examined the expression of PAkt, α-SMA and E-cadherin by western blotting to investigate the influence of Wortmannin, the inhibitor of PI3K, after HPMC promoted by TGFβ1. 【Results】The significantly increase of p-Akt showed up at 0.5 h after induction of TGFβ1, reached the summit at 1 h and decreased gradually. The expression of α-SMA and E-cadherin are markedly changed increased and decreased respectively after 48 h incubation with 10 ng/ml TGFβ1; However, the expression of p-Akt and α-SMA reversed significantly when incubated with Wortmannin and TGFβ1 simultaneously. 【Conclusion】PI3K/Akt signal system involves the process of epithelia-mesenchymal transition of human peritoneal mesothelial cell induced by transforming growth factor β1 and the inhibitor of the pathway may prevent peritoneal fibrosis effectively.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2010年第5期672-675,共4页
China Journal of Modern Medicine
基金
湖南省科技厅项目(No:2007SK3041)
