地龙提取物通过抑制TGF-β1信号通路发挥抗肺纤维化作用研究

目的:探讨地龙提取物对抑制转化生长因子-β1(TGF-β1)信号通路诱导的肺泡上皮细胞间质转化和肺成纤维细胞活化的影响。方法:MTT法分析地龙提取物对两种细胞增殖的影响。TGF-β1诱导A549肺泡上皮细胞的间质转化和MRC-5肺成纤维细胞的活化; 圆形度定量法分析细胞形态,免疫细胞化学法检测细胞E-钙黏附素(E-cad)和Collagen-Ⅰ蛋白的表达; 双抗体夹心酶联免疫吸附法检测Collagen-Ⅰ和FN水平。结果:对于A549细胞,与正常组比较,TGF-β1组圆形度和E-cad表达均显著降低(P<0.01),Collagen-Ⅰ表达显著升高(P<0.01); 与TGF-β1组比较,地龙组能显著抑制E-cad表达的下调(P<0.05),地龙组能显著抑制Collagen-Ⅰ表达的上调(P<0.05)。对于MRC-5细胞,与正常组比较,TGF-β1组FN、Collagen-Ⅰ的分泌量均显著增加(P<0.01); 与TGF-β1组比较,地龙组的上述两种蛋白的分泌量均显著减少(P<0.05)。结论:地龙提取物抑制TGF-β1诱导的肺泡上皮细胞的间质转化和肺成纤维细胞的活化,提示地龙提取物可能具有抑制肺纤维化的形成和发展的作用。

Analysis on the antifibrotic effect of earthworm extract by inhibiting TGF-β1 signaling pathway

Objective:To investigate the effect of earthworm extract on inhibition of stromal transformation of alveolar epithelial cells induced by TGF-β1 signaling pathway and activation of lung fibroblasts.Methods:MTT method was used to analyze the effect of earthworm extract on the proliferation of two kinds of cells.TGF-β1 induced mesenchymal transformation of A549 alveolar epithelial cells and activation of mrc-5 lung fibroblasts by TGF-β1.The cell morphology was analyzed by circular quantitative method,and the expression of e-cad and Collagen-Ⅰprotein was detected by immunocytochemistry.Collagen-Ⅰand FN levels were detected by double antibody sandwich enzyme-linked immunosorbent assay.Results:Compared with the normal group,the expression of Collagen-I and TGF-β1 in A549 cells was significantly decreased(P<0.01),and the expression of Collagen-Ⅰwas significantly increased(P<0.01).Compared with TGF-β1 group 1,earthworm group could significantly inhibit the down-regulation of e-cad expression(P<0.05),while earthworm group could significantly inhibit the up-regulation of Collagen-Ⅰexpression(P<0.05).For mrc-5 cells,compared with the normal group,the secretion levels of FN and Collagen-Ⅰin TGF-β1 group increased significantly(P<0.01).Compared with TGF-β1 group,the secretion of the two proteins in earthworm group was significantly reduced(P<0.05).Conclusion:Earthworm extract inhibits the stromal transformation of alveolar epithelial cells induced by TGF-β1 and the activation of lung fibroblasts,suggesting that earthworm extract may inhibit the formation and development of pulmonary fibrosis.