白藜芦醇抑制TGF-β1/Smad3信号通路对人肺上皮细胞A549 EMT调控的影响及机制  

Study of the effects and the mechanisms of resveratrol on the regulation of human pulmonary epithelial cell A549 by inhibiting TGF-β1/Smad3 signaling pathway

作　　者：张云珍 方海燕 余红[3] 饶珊珊[3] 石喆[1] 邵松军[3] ZHANG Yun-zhen;FANG Hai-yan;YU Hong;RAO Shan-shan;SHI Zhe;SHAO Song -jun(Department of Respiratory Medicine, the First People's Hospital of Huizhou, Huizhou516000, Guangdong, China)

机构地区：[1]惠州市第一人民医院呼吸内科,广东惠州516000 [2]贵州省第二人民医院老年精神科,贵州贵阳550004 [3]贵州省人民医院呼吸与重症医学科,贵州贵阳550002

出　　处：《广东医学》2019年第15期2130-2134,共5页Guangdong Medical Journal

基　　金：贵州省科技计划项目[编号:黔科合基础[2018]1098];贵州省卫生计生委科学技术基金项目(编号:gzwjkj2017-1-019);贵阳市科技计划项目(编号:筑科合同[20161001]09号)

摘　　要：目的探讨白藜芦醇(resveratrol,Res)对TGF-β1诱导的人肺上皮细胞(A549细胞株)上皮间质转化(epithelial-mesenchymal transition,EMT)的抑制作用及其机制。方法本实验设立对照组、转化生长因子-β1(transforming growth factor,TGF-β1)组(TGF-β1组)、TGF-β1+Res组(TGF-β1+Res组)。甲基偶氮唑盐比色法(MTT)确定白藜芦醇的干预浓度;蛋白免疫印迹法(Western blot)测定不同组细胞蛋白的表达变化;实时荧光定量PCR(RT-qPCR)检测钙黏蛋白E(E-cadherin)和α-平滑肌肌动蛋白(alpha smooth muscle actin,α-SMA)mRNA的表达。结果 TGF-β1刺激后,与对照组相比,A549细胞的增殖能力明显增强;总的Smad3蛋白(t-Smad3)表达无变化,但磷酸化的Smad3蛋白(p-Smad3)表达增多;上皮细胞标志蛋白E-cadherin表达减少(P<0.05),间质细胞标志蛋白α-SMA表达明显增多(P<0.05);白藜芦醇干预后,t-Smad3蛋白表达几乎无变化,p-Smad3蛋白表达减少;E-cadherin蛋白表达增加(P<0.05),α-SMA蛋白表达减少(P<0.05);RT-qPCR的检测结果与Western blot结果一致。结论白藜芦醇通过阻断TGF-β1/Smad3信号通路转导,抑制了TGF-β1诱导的人肺上皮细胞A549的增殖及上皮间质转化。Objective To investigate resveratrol(Res) inhibiting the epithelial-mesenchymal transition(EMT) of human pulmonary epithelial cells(A549 cell line) induced by TGF-β1 and its mechanism. Methods This experiment established three cell groups: control group, TGF-β1 group, and TGF-β1+Res group. The intervention concentration of resveratrol was determined by MTT assays. The expression of proteins in different group was assessed by Western blot. RT-qPCR was used to assess the mRNA expression of E-cadherin and α-SMA. Results After TGF-β1 stimulation, the proliferation capacity of A549 cells was significantly enhanced compared with the control group. The total Smad3 protein(t-Smad3) expression did not change, but the phosphorylated Smad3(p-Smad3) protein expression was increased. The expression of the epithelial marker E-cadherin was reduced(P<0.05), and the expression of the mesenchymal marker α-SMA was significantly increased(P<0.05). When resveratrol treatment, the total Smad3 protein expression was almost unchanged, and the phosphorylated Smad3(p-Smad3) protein expression was reduced. E-cadherin protein expression increased(P<0.05), while α-SMA protein expression decreased(P<0.05). The results of RT-qPCR were consistent with the results of Western blot.Conclusion Resveratrol inhibits the proliferation and epithelial mesenchymal transformation of human pulmonary epithelial A549 cells through blocking TGF-β1/Smad3 signaling pathway transduction.

关 键 词：白藜芦醇 A549细胞 上皮间质转化 TGF-β1/Smad3信号通路 

分 类 号：R73-3[医药卫生—肿瘤] R734.2[医药卫生—临床医学]