羊栖菜多糖对TGF-β1诱导的大鼠肺成纤维细胞的影响

摘要

目的 研究羊栖菜多糖对TGF-β1诱导的大鼠肺成纤维细胞的影响,探讨其抗肺纤维化作用及机制.方法 将实验分成对照组、TGF-β1组、TGF-β1+不同剂量羊栖菜多糖组(25、50、100μg/ml),采用MTT法检测细胞增殖,RT-PCR法检测Smad3、Smad7、α-SMA、Ⅰ型胶原mRNA的表达,ELISA法检测Ⅰ型胶原蛋白的表达,Western blot法检测α-SMA的表达.结果 TGF-β1能诱导肺成纤维细胞增殖及表达Smad3 mRNA、Smad7 mRNA、Ⅰ型胶原、α-SMA(P ＜0.05).羊栖菜多糖可以不同程度抑制TGF-β1诱导的肺成纤维细胞的增殖以及Smad3mRNA、Ⅰ型胶原和α-SMA的表达,且呈剂量依赖性(P＜0.05),能促进TGF-β1诱导的肺成纤维细胞表达Smad7 mRNA,呈剂量依赖性(P＜0.05).结论 在离体细胞,羊栖菜多糖能抑制TGF-β1诱导的肺成纤维细胞增殖、分化与表达,其机制可能与其下调Smad3,促进Smad 7表达从而抑制TGF-β/Smads信号通路有关.%Objective To investigate the effects of Sargassum fusiforme polysaccharide to the transforming growth factor beta1 (TGF-β1)-induced rat lung fibroblast cells in vitro,and to evaluate the anti-fibrotic mechanism of SFPS.Methods Cultured rat lung fibroblast cells were divided into a control group,a TGF-β1-treated group and TGFβ1 (5ng/ml) plus different dosage SFPS-treated groups(TGF-β1 5ng/ml + SFPS 25μg/ml group;TGF-β1 5ng/ml + SFPS 50μg/ml group;TGF-β1 5ng/ml + SFPS 100μg/ml group).Cell proliferation was assessed by MTT colorimetric assay.The levels of collagen type Ⅰ mRNA and protein was measured by RT-PCR and ELISA.RT-PCR and Western blot were used to detect the expression of α-smooth muscle actin mRNA and protein.RT-PCR were used to detect the expression of Smad3,Smad7mRNA.Results TGF-β1 induced proliferation and the expressions of Smad3,Smad7mRNA,collagen type Ⅰ,α-smooth muscle actin of lung fibroblasts (P ＜ 0.05).SFPS prevented TGF-β1-induced proliferation and the expressions of Smad3mRNA,collagen type Ⅰ,α-smooth muscle actin of lung fibroblasts,this prevention effect was dose-dependent (P ＜ 0.05).SFPS increased TGF-β1-induced expressions of Smad7 mRNA this effect was also dose-dependent (P ＜ 0.05).Conclusion In vitro cell,SFPS can inhibit TGF-β1-induced lung fibroblast proliferation,differentiation and expression.The mechanism may be related to reduced Smad3,promoted Smad7 thereby inhibiting TGF-β/Smads signaling pathway.