黄芪甲苷调控Akt信号通路阻抑人腹膜间皮细胞间充质转化的实验研究
The Inhibitory Effect of Astragaloside Ⅳ on Epithelial Mesenchymal Transition of Human Peritoneal Mesothelial Cells by Regulating Akt Signaling
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摘要: 目的 观察黄芪甲苷（AS-Ⅳ）对TGF-β1诱导人腹膜间皮细胞HMrSV5 EMT及β-catenin的影响，探讨Akt信号通路的作用及AS-Ⅳ干预机制。方法 采用TGF-β1诱导建立HMrSV5 EMT模型，Western blot检测不同浓度TGF-β1对EMT标记蛋白、β-catenin及Akt信号蛋白的影响；予不同浓度AS-Ⅳ干预HMrSV5 EMT模型，Real-time PCR检测EMT相关基因及β-catenin mRNA水平，Western blot检测Akt信号蛋白表达；与Akt通路抑制剂MK2206、雷帕霉素及激动剂Insulin作对照，观察AS-Ⅳ干预HMrSV5 EMT指标及β-catenin蛋白的变化。结果 ①TGF-β1诱导HMrSV5细胞发生EMT、上调β-catenin水平，激活Akt信号通路；②AS-Ⅳ能不同程度改善TGF-β1诱导的HMrSV5 EMT、降解β-catenin，并在一定程度上抑制Akt信号通路活化；③Akt通路参与调控HMrSV5 EMT及β-catenin，其抑制剂MK2206、雷帕霉素的调控作用与AS-Ⅳ类似；④Akt通路激动剂Insulin明显减弱AS-Ⅳ对EMT及β-catenin的抑制效果。结论 TGF-β1可诱导HMrSV5 EMT，上调β-catenin，AS-Ⅳ可能通过抑制Akt信号通路活化，阻抑HMrSV5 EMT，降解β-catenin。Abstract: OBJECTIVE To observe the effect of Astragaloside Ⅳ (AS-Ⅳ) on EMT of human peritoneal mesothelial cells (HPMCs) HMrSV5， exploring the function of Akt signaling and regulatory mechanism of AS-Ⅳ. METHODS We used TGF-β1 to establish EMT model of HMrSV5. EMT marker proteins， β-catenin and Akt signaling-related proteins were detected by Western blotting analysis. β-catenin and EMT-related mRNA were expressed by Real-time PCR analysis. Compared with MK2206， Rapamycin and Insulin， the effect of AS-Ⅳ on EMT and β-catenin was observed by Western blotting. RESULTS ①The EMT model of HMrSV5 was established by TGF-β1. TGF-β1 increased β-catenin expression and activated Akt signaling. ②AS-Ⅳ could alleviate TGF-β1-treated EMT of HMrSV5， decrease β-catenin expression， as well as inhibit activation of Akt signaling. ③EMT of HMrSV5 and β-catenin were regulated by Akt signaling， MK2206 and Rapamycin had the similar effect with AS-Ⅳ. ④Insulin could reduce obviously inhibitory effect on EMT and β-catenin of AS-Ⅳ. CONCLUSION TGF-β1 can induce EMT of HMrSV5 and upregulate β-catenin， AS-Ⅳ has inhibitory effect on EMT and β-catenin of HMrSV5 by regulating Akt signaling.
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