吗替麦考酚酯对肺成纤维细胞的转化及功能影响

发布时间：2019-01-08 14:16

【摘要】：目的:通过吗替麦考酚酯(MMF)对体外培养肺成纤维细胞(LF)转化为肌成纤维细胞(MF)过程的干预,观察MMF对LF转化及CTGF、FN蛋白表达的影响。方法:体外培养新生大鼠来源的LF,加入转化生长因子-β1(TGF-β1)诱导转化为MF,分别加入终浓度为0μmol/L(对照组)、0.1μmol/L(低浓度组)、1μmol/L(中浓度组)、10μmol/L(高浓度组)的MMF进行干预;倒置显微镜下观察LF和MF形态,并用细胞免疫荧光的方法鉴定波形蛋白和α-平滑肌肌动蛋白(α-SMA),分析MMF对LF转化的影响;ELISA方法检测结缔组织生长因子(CTGF)和纤维连接蛋白(FN)的水平。结果:LF经TGF-β1诱导96 h后转化成MF,细胞免疫荧光结果显示MMF能够抑制α-SMA的表达,但对已分化的肌成纤维细胞表型变化无影响;ELISA结果显示与对照组比较,MMF组CTGF和FN分泌水平均显著下降(P0.05),并呈一定的浓度依赖。结论:MMF能够抑制肺成纤维细胞转化和CTGF、FN表达,提示MMF有抗纤维化作用,影响CTGF和FN表达可能是作用途径之一。
[Abstract]:Aim: to investigate the effect of mycophenolate mofetil (MMF) on LF transformation and CTGF,FN protein expression in cultured lung fibroblasts by interfering with the process of transforming (LF) into myofibroblasts (MF) in vitro. Methods: LF, derived from neonatal rats were induced by transforming growth factor- 尾 1 (TGF- 尾 1) into MF, with final concentration of 0 渭 mol/L (control group), 0.1 渭 mol/L (low concentration group) and 1 渭 mol/L (medium concentration group), respectively. The MMF of 10 渭 mol/L (high concentration group) was interfered with. The morphologies of LF and MF were observed under inverted microscope. Vimentin and 伪-smooth muscle actin (伪-SMA),) were identified by immunofluorescence. The effect of MMF on the transformation of LF was analyzed. The levels of connective tissue growth factor (CTGF) and fibronectin (FN) were detected by ELISA. Results: LF was induced by TGF- 尾 1 for 96 h and transformed into MF, cells. The results of immunofluorescence showed that MMF could inhibit the expression of 伪-SMA, but had no effect on the phenotypic changes of differentiated myofibroblasts. The results of ELISA showed that the levels of CTGF and FN in MMF group were significantly lower than those in control group (P0.05), and in a certain concentration dependent manner. Conclusion: MMF can inhibit the transformation of pulmonary fibroblasts and the expression of CTGF,FN, suggesting that MMF has anti-fibrosis effect, and the influence of CTGF and FN expression may be one of the mechanisms.
【作者单位】：徐州医科大学附属医院风湿免疫科;
【基金】：徐州市科技项目(KC16SH111)基金资助
【分类号】：R563

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