EZH2对HSC-T6细胞增殖活化的影响及其部分机制研究

发布时间：2018-02-08 14:44

本文关键词： 组蛋白甲基化肝星状细胞 Zeste基因增强子同源物 DZNep 肝纤维化 p-ERK p-AKT α平滑肌肌动蛋白　出处：《中国药理学通报》2015年08期 　论文类型：期刊论文

【摘要】：目的探讨Zeste基因增强子同源物2(enhancer of zeste homolog 2,EZH2)表达沉默对大鼠肝星状细胞HSC-T6细胞增殖活化的影响,及其部分调控机制。方法应用EZH2的抑制剂DZNep(3-Deazaneplanocin A)作用于TGF-β1诱导活化的HSC-T6细胞,采用Western blot法检测蛋白EZH2、p-ERK、p-AKT和α-SMA的表达;根据EZH2的碱基序列设计并合成小干扰RNA(small interfering RNA,si RNA),通过脂质体LipofectamineTM2000转染到HSC-T6细胞内,应用四甲基偶氮唑盐(MTT)法检测HSC-T6细胞的增殖变化,Western blot法检测蛋白EZH2、p-ERK、p-AKT和α-SMA的表达。结果将DZNep加入TGF-β1诱导活化的HSC-T6细胞后,EZH2蛋白水平明显降低,同时p-ERK、pAKT和α-SMA蛋白水平亦明显降低;将EZH2-si RNA转染活化的HSC-T6细胞内,HSC-T6细胞的增殖可明显被抑制,同时EZH2、p-ERK、p-AKT和α-SMA蛋白水平亦明显降低。结论抑制EZH2的表达可明显抑制HSC-T6细胞的增殖活化,EZH2可能是潜在的治疗肝纤维化的靶点。
[Abstract]:Objective to investigate the effect of silencing the expression of Zeste enhancer of zeste homolog 2 (EZH2) on the proliferation and activation of HSC-T6 cells in rat hepatic stellate cells, and its mechanism. Methods DZNep(3-Deazaneplanocin A, an inhibitor of EZH2, was used to induce the activation of HSC-T6 cells induced by TGF- 尾 1. Western blot method was used to detect the expression of EZH2p-ERKG p-AKT and 伪 -SMA, and the small interfering RNA(small interfering RNA-siRNAs were designed and synthesized according to the base sequence of EZH2, and were transfected into HSC-T6 cells by liposome LipofectamineTM2000. The proliferative changes of HSC-T6 cells were detected by tetramethylazolazolium (MTT) assay and the expression of EZH2pERKnp-AKT and 伪 -SMA were detected by Western blot. Results the level of EZH2 protein in activated HSC-T6 cells induced by TGF- 尾 1 was significantly decreased, and the protein levels of p-ERKPAKT and 伪 -SMA were also significantly decreased when DZNep was added to the activated HSC-T6 cells. The proliferation of HSC-T6 cells transfected with EZH2-si RNA was significantly inhibited. At the same time, the levels of EZH _ 2 p-ERK _ (p-AKT) and 伪 -SMA protein were also significantly decreased. Conclusion inhibiting the expression of EZH2 can significantly inhibit the proliferation and activation of HSC-T6 cells. EZH _ 2 may be a potential target for the treatment of liver fibrosis.
【作者单位】：安徽医科大学药学院;安徽医科大学肝病研究所安徽省高校产业共性技术研究院;
【基金】：国家自然科学基金资助项目(No 81273526) 博士点基金资助项目(No 20123420120001) 安徽省教育厅基金资助项目(No KJ2012A156) 安徽省自然科学基金资助项目(No 1308085MH145) 安徽医科大学基金资助项目(No XJ201118)
【分类号】：R575.2

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