柴胡皂甙D通过调控TGF-β1/Smads信号通路抑制人胚肺成纤维细胞增殖和胶原蛋白产生  

Saikosaponin D inhibits proliferation and collagen production of human embryonic lung fibroblasts by regulating TGF-β1/Smads signaling pathway

作　　者：孙金玲 郑金旭[1] 史小东 钱海[1] 金承基 徐莉莉 SUN Jinling;ZHENG Jinxu;SHI Xiaodong;QIAN Hai;JIN Chengji;XU Lili(Hospital Affiliated to Jiangsu University, Zhenjiang 212000, China)

机构地区：[1]江苏大学附属医院,江苏镇江212000

出　　处：《细胞与分子免疫学杂志》2019年第3期256-261,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：镇江市重点研发计划(社会发展)项目(SH2018048)

摘　　要：目的研究柴胡皂甙D(SSD)对转化生长因子β1(TGF-β1)诱导的人胚肺成纤维细胞(HELF)增殖、转分化以及TGF-β1/果蝇抗生物皮肤生长因子蛋白家族(Smads)信号通路的影响。方法体外培养HELF,设立正常对照组、 1 ng/mL TGF-β1处理组、 1 ng/mL TGF-β1联合0.5μmol/L SSD处理组、 1 ng/mL TGF-β1联合1μmol/L SSD处理组、 1 ng/mL TGF-β1联合2μmol/L SSD处理组。采用CCK-8法检测HELF的增殖情况,荧光定量PCR法检测Smad2、 Smad3、 Smad7的mRNA水平, Western blot法检测α平滑肌肌动蛋白(α-SMA)、 1型胶原蛋白(Col1)、 Smad2、 Smad3、磷酸化的Smad2(p-Smad2)、 p-Smad3、 Smad7的蛋白水平。结果与正常对照组比较, TGF-β1处理组细胞增殖明显, Col1、α-SMA蛋白水平增加, Smad2和Smad3 mRNA和蛋白磷酸化水平显著增加, Smad7 mRNA和蛋白水平显著降低;与TGF-β1处理组比较,不同浓度SSD处理组细胞增殖降低,可逆转以上各指标的改变,呈一定的量效关系。结论 SSD通过调控TGF-β1/Smads信号通路,发挥抗肺纤维化的作用。Objective To investigate the effect of saikosaponin D(SSD) on the proliferation and transformation of human embryonic lung fibroblasts(HELFs) induced by transforming growth factor-beta 1(TGF-β1) and the regulation of signal pathway of TGF-β1/Smads family. Methods HELFs were cultured in vitro and divided into 5 groups: a control group, 1 ng/mL TGF-β1-induced group, 1 ng/mL TGF-β1 combined with 0.5 μmol/L SSD treatment group, 1 ng/mL TGF-β1 combined with 1 μmol/L SSD treatment group, and 1 ng/mL TGF-β1 combined with 2 μmol/L SSD treatment group. Cell viability of HELFs was detected by CCK-8 assay. The expression of Smad2, Smad3 and Smad7 mRNA were detected by real-time fluorescence quantitative PCR. The protein levels of α-smooth muscle actin(α-SMA), type 1 collagen(Col1), Smad2, Smad3, phosphorylated Smad2(p-smad2), p-smad3 and Smad7 were assessed by Western blot analysis. Results Compared with the control group, TGF-β1-induced group showed the apparently increased proliferation ability, the increased protein levels of Col1 and α-SMA, the significantly increased mRNA and protein phosphorylation levels of Smad2 and Smad3, and the significantly decreased mRNA and protein expression of Smad7. Compared with the TGF-β1-induced group, the cell proliferation of HELFs in different concentrations of SSD treatment groups was reduced, which could reverse the changes of the above indicators in a dose-dependent manner. Conclusion SSD plays an important role in anti-pulmonary fibrosis by regulating TGF-β1/Smads signaling pathway.

关 键 词：柴胡皂甙D 人胚肺成纤维细胞(HELF) 转化生长因子β1(TGF-β1) 果蝇抗生物皮肤生长因子蛋白(Smads) 

分 类 号：R392-33[医药卫生—免疫学] R285.5[医药卫生—基础医学]