丹参酮ⅡA对大鼠肺纤维化细胞TGF-β1/Smads信号通路的影响  

Effect of Tanshinone ⅡA on TGF-β1/Smads Signaling Pathway in Pulmonary Fibrosis Cellsof Rats

作　　者：邵笑[1] 祝骥[1] 赵峻峰[1] 许颖龄[2] 张颖颖[2] 杨建清[2] 卢德赵[2] 

机构地区：[1]浙江中医药大学附属第三医院检验科,杭州310005 [2]浙江中医药大学生命科学学院,杭州310053

出　　处：《浙江中西医结合杂志》2016年第5期414-417,共4页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金项目(No.81102841;81403133);浙江省自然科学基金项目(No.LY13H270005;No.LQ14H280004);中国博士后科学基金(No.2013M541800)

摘　　要：目的观察丹参酮ⅡA对TGF-β1诱导大鼠肺成纤维细胞TGF-β1/Smads信号通路的影响,探讨丹参酮ⅡA干预肺纤维化的机制。方法体外培养大鼠肺成纤维细胞(HFL-1),选取生长状态良好的对数期细胞分为正常组、TGF-β1组及丹参酮ⅡA低(L)、中(M)、高(H)剂量组。除正常组外,余四组予以10ng/m L TGF-β1诱导HFL-1纤维化24h,同时丹参酮ⅡA低、中、高剂量组分别予2μmol/L、5μmol/L和10μmol/L丹参酮ⅡA处理24h,ELISA法测定各组细胞中TGF-β1受体表达,荧光定量PCR法及免疫印迹法检测Smad2、Smad7表达情况。结果与正常组比较,HLF-1细胞经10ng/m L TGF-β1诱导后细胞TGF-β1受体和Smad2表达量增加[(0.19±0.23)比(1.41±0.12)、(1.00±0.12)比(1.90±0.12),P<0.01],Smad7表达量降低[(1.00±0.15)比(0.57±0.09),P<0.01];与TGF-β1组比较,10μmol/L丹参酮ⅡA处理后TGF-β1受体和Smad2的表达量均有下降趋势[(1.90±0.23)比(1.53±0.12),(1.90±0.12)比(1.38±0.06),P<0.01],Smad7表达量升高[(0.57±0.08)比(0.74±0.11),P<0.01]。结论丹参酮ⅡA可以抑制TGF-β1诱导的大鼠肺成纤维细胞增殖和Smad2及TGF-β1受体表达,增强抑制性信号蛋白Smad7的表达,从而发挥抑制肺纤维化的作用。Objective To investigate the protection effect of tanshinone ⅡA on transform growth factor（TGF）-β1/Smads pathway in pulmonary fibrosis cells of rats. Methods HLF-1 cells were cultivated in vitro and induced to pulmonary fibrosis cells with 10ng/m L TGF-β1, then the cells were treated with 2μmol/L, 5μmol/L and 10μmol/L tanshinoneⅡA, respectively, for 24 h. The method of Elisa was used to measure the expression of TGF-β1 receptors. The expression of Smad 2 and Smad 7 was determined with the method of fluorogenic quantitative PCR and Western Blotting. Results 10ng/m L TGF-β1increased the expression of TGF-β1 receptors（1.41±0.12 vs 0.19±0.23） and Smad 2（1.90±0.12 vs 1.00±0.12）, and decreased the expression of Smad 7（0.57±0.09 vs 1.00±0.15） in the HLF-1 cells（all P〈0.05）. When the cells were treated withtanshinone ⅡA, TGF-β1 receptors（1.53±0.12） and Smad 2（1.38±0.06） expressed with a downward trend while the expression of Smad 7（0.74±0.11） was increased（all P〈0.05）. Conclusion TGF-β1 can promote pulmonary fibrosis through upregulating the expression of TGF-β1receptors and Smad 2 and downregulatingthe expression of Smad 7. Tanshinone Ⅱ A can regulate the TGF-β1/Smads signaling pathway, which plays a preventive role in pulmonary fibrosis.

关 键 词：大鼠 肺纤维化 HFL-1 丹参酮ⅡA 转化生长因子Β 

分 类 号：R285.5[医药卫生—中药学]