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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 effect of the extract of Cortex Moutan (a composition of paeonol, total glycosides and polysaccharides in a certain ratio) on TGF-β1-induced interstitial transformation of alveolar epithelial cells and activation of lung fibroblasts. Methods: The effects of Cortex Moutan on the proliferation of two kinds of cells were analyzed by MTT assay. TGF-β1 induced A549 alveolar epithelial cell stromal transformation and MRC-5 lung fibroblast activation, with different doses of moutan intervention; circularity quantitative analysis of cell morphology, immunocytochemistry detection of cell E-cad and Collagen-I protein expression; double antibody sandwich enzyme-linked immunosorbent assay Collagen-I and FN levels. Results: There was no significant difference in the proliferation of A549 cells and MRC-5 cells between 12.5 ~ 200 μg / m L Cortex Moutan (P> 0.05). In A549 cells, the expression of E-cad and circularity of TGF-β1 group was significantly lower than that of normal group (P <0.01), while Collagen-I expression was significantly increased (P <0.01). Compared with TGF-β1 group, However, 50 ~ 200μg / m L Cortex Moutan significantly inhibited the down-regulation of E-cad (P <0.05 or 0.01), and 200μg / m L Cortex Moutan significantly inhibited the changes of circularity (P> 0.05) Collagen-I expression was up-regulated (P <0.05). For MRC-5 cells, the secretion of FN and Collagen-I in TGF-β1 group was significantly increased compared with the normal group (P <0.01). Compared with the TGF-β1 group, the extraction of 100 μg / ml and 200 μg / The amount of secretion of the above two proteins in the subject group was significantly reduced (P <0.05 or 0.01). CONCLUSION: The extract of Cortex Moutan inhibits the mesangial transformation of alveolar epithelial cells induced by TGF-β1 and the activation of lung fibroblasts, suggesting that the stem may inhibit the formation and development of pulmonary fibrosis.