牡丹皮提取物抑制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

Extracts from Cortex Moutan Inhibits Epithelial to Mesenchymal Transition of Alveolar Epithelial Cells and Activation of Pulmonary Fibroblasts

目的：探讨牡丹皮提取物（丹皮酚、总苷和多糖提取物按照一定比例形成的组合物）对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的影响。方法：MTT法分析牡丹皮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,用不同剂量的牡丹皮干预;圆形度定量法分析细胞形态,免疫细胞化学法检测细胞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：12.5-200μg/m L牡丹皮对A549细胞和MRC-5细胞的增殖没有显著性影响（P〉0.05）。对于A549细胞,与正常组比较,TGF-β1组圆形度和E-cad表达显著降低（P〈0.01）,Collagen-I表达显著升高（P〈0.01）;与TGF-β1组比较,牡丹皮不能显著抑制圆形度的变化（P〉0.05）,但50-200μg/m L牡丹皮能显著抑制E-cad表达的下调（P〈0.05或0.01）,200μg/m L牡丹皮能显著抑制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的分泌量显著增加（P〈0.01）;与TGF-β1组比较,100μg/m L和200μg/m L牡丹皮提取物组的上述两种蛋白的分泌量显著减少（P〈0.05或0.01）。结论：牡丹皮提取物抑制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,提示牡丹皮可能具有抑制肺纤维化的形成和发展的作用。

Objective： To investigate the inhibitory effect of cortex Moutan extracts on alveolar epithelial to mesenchymal transition and activation of pulmonary fibroblast.Methods： MTT colorimetric method was applied for evaluation of cortex Moutan extracts on proliferation of both A549 cells,a type of alveolar epithelial cell line and MRC-5 cells,a type of pulmonary fibroblasts.And then,TGF-β1 was used to induce epithelial to mesenchymal（EMT） of A549 cells and induce activation of pulmonary fibroblasts.Simultaneously,various doses of cortex Moutan extracts were treated the cells.Cellular roundness was applied to quantitative evaluation of cellular morphological change.Immunocytochemistry was applied to examine E-cadherin and collagen-I proteins expression in the cells.Enzyme linked-immuno-sorbent assay（ELISA） was applied to Collagen-I and FN levels.Results： The cortex Moutan extracts with the dose of 12.5 - 200 μg/m L had no significant effect on cells proliferation.EMT of A549 cells was induced by TGF-β1,showing significant change of roundness,E-cadherin and collagen-I（P〈0.01）.Compared to TGF-β1,cortex Moutan extracts did not significantly change the roundness（P〉0.05）; however,50 - 200 μg/m L cortex Moutan extracts increased E-cadherin significantly（P〈0.05 or P〈0.01） and 200 μg/m L cortex Moutan extracts decreased collagen-I（P〈0.05）.Furthermore,the activation of MRC-5 cells was induced by TGF-β1,showing significant increase of FN and collagen-I（P〈0.01）.Compared to TGF-β1,100 μg/mL and 200 μg/mL cortex Moutan extracts could significantly decrease the two proteins levels（P〈0.05 or P〈0.01）.Conclusion： Cortex Moutan extracts could inhibit alveolar EMT and pulmonary fibroblast activation induced by TGF-β1,showing inhibition for pulmonary fibrosis.