柴胡皂甙D经mTOR通路抑制A549细胞的上皮间质转化  

Saikosaponin D inhibits epithelial mesenchymal transition of A549 cells via mTOR signaling pathway

作　　者：史小东 郑金旭[1] 钱海[2] 杨磊 孙金玲 SHI Xiao-dong;ZHENG Jin-xu;QIAN Hai;YANG Lei;SUN Jin-ling(Department of Respiratory Medicine,the Affiliated Hospital of Jiangsu University,Zhenjiang Jiangsu 212001;School of Medicine, Jiangsu University,Zhenjiang Jiangsu 212013,China)

机构地区：[1]江苏大学附属医院呼吸内科,江苏镇江212001 [2]江苏大学医学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2019年第2期113-116,共4页Journal of Jiangsu University：Medicine Edition

基　　金：镇江市社会发展项目(SH2018048)

摘　　要：目的:探讨柴胡皂甙D(SSd)对人肺腺癌A549细胞上皮间质转化(epithelial mesenchymal transition,EMT)的影响及可能机制。方法:体外培养A549细胞,经TGF-β1 (10 ng/m L)刺激诱导EMT,观察不同浓度SSd(0.5,1.0,2.0μmol/L)的抑制作用。采用MTT法检测细胞增殖活性,蛋白质印迹检测A549细胞mTOR、p70S6、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、N-钙黏蛋白、Ⅰ型胶原蛋白、E-钙黏蛋白的表达水平。结果:MTT检测结果显示,TGF-β1刺激后A549细胞增殖能力增强,SSd可明显抑制TGF-β1刺激的细胞增殖能力,且呈剂量依赖关系。经SSd作用后,TGF-β1刺激的A549细胞mTOR及下游的p70S6蛋白表达明显下调,α-SMA、Ⅰ型胶原蛋白、N-钙黏蛋白的表达水平明显降低,而E-钙黏蛋白则明显升高,且呈明显的剂量—时间依赖关系。结论:SSd可通过mTOR信号通路抑制A549细胞EMT过程。Objective: To study the effect and mechanism of saikosaponin D ( SSd) on transforming growth factor-β1( TGF-β1) induced epithelial mesenchymal transition ( EMT) in A549 cells. Methods: A549 cells were cultured in vitro,and EMT was induced by TGF-β1 ( 10 ng /mL). The inhibitory effects of different concentrations of SSd( 0. 5,1. 0,2. 0 μmol /L) were observed. Cell proliferation activity was analyzed by MTT assay,protein expression levels of mTOR,p70S6,α-smooth muscle actin (α-SMA), N-cadherin,type Ⅰ collagen( collagen Ⅰ) and E-cadherin were detected by Western blotting. Results: The proliferation of A549 cells stimulated by TGF-β1 was enhanced,and SSd significantly reversed the proliferation of TGF-β1-stimulated cells in a dose-dependent manner. After treatment with SSd,the expression of mTOR and downstream p70S6 protein were significantly decreased in TGF-β1 stimulated A549 cells,and the expression levels of α-SMA,collagen Ⅰ and N-cadherin were significantly decreased, while E-cadherin was significantly increased in the dose-and time-dependent manner. Conclusion: SSd can inhibit A549 cells EMT via a mTOR signaling pathway.

关 键 词：柴胡皂甙D MTOR A549 细胞 上皮间质转化 转化生长因子-Β1 

分 类 号：R285.5[医药卫生—中药学]