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Smad 3、Smad 7、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.
出处
《医学研究杂志》
2017年第4期74-77,共4页
Journal of Medical Research
基金
浙江省卫生厅基金资助项目(2014KYB302)
作者简介
通讯作者：张伟文，主任医师，电子信箱：ZWW5641@126．com
