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目的:探讨蛇床子素对转化生长因子-β1(TGF-β1)诱导的小鼠肺成纤维细胞(MLF)增殖、胶原合成和表型转化的影响并探讨机制。方法:使用乳酸脱氨酶(LDH)试剂盒、细胞增殖-毒性(CCK-8)试剂盒、细胞计数法、~3H-脯氨酸掺入实验检测各组细胞的死亡、增殖、胶原合成能力;Western blot技术检测胶原Ⅰ、胶原Ⅲ、Rac-1蛋白表达水平。细胞免疫荧光检测波形蛋白(vimentin)、α-平滑肌肌动蛋白(α-SMA)在细胞内的定位及表达水平。DCFH-DA检测细胞内活性氧水平。结果:0.1-50μmol/L的蛇床子素对MLF没有毒性。10-50μmol/L的蛇床子素能够浓度依赖性地抑制TGFβ-1诱导的MLF增殖、胶原合成、α-SMA、Rac-1蛋白表达升高及ROS生成。结论:蛇床子素可以浓度依赖性地抑制TGFβ-1诱导的肺成纤维细胞的增殖、胶原蛋白合成和表型分化,其可能机制与抑制Rac-1表达、活性氧(ROS)生成有关。
OBJECTIVE: To investigate the effects of osthole on the proliferation, collagen synthesis and phenotype of lung fibroblasts (MLF) induced by transforming growth factor-β1 (TGF-β1) in mice and its mechanism. Methods: LDH, CCK-8, cell counting and 3H-proline incorporation assay were used to detect the cell death, proliferation and collagen synthesis in each group Western blot was used to detect the expression of collagen Ⅰ, collagen Ⅲ and Rac-1. Cell immunofluorescence was used to detect the expression of vimentin and α-smooth muscle actin (α-SMA) in the cells. DCFH-DA detection of intracellular reactive oxygen species. Results: 0.1-50μmol / L osthole was not toxic to MLF. Opihonol at concentration of 10-50μmol / L inhibited MLF proliferation, collagen synthesis, α-SMA, Rac-1 protein expression and ROS production induced by TGFβ-1 in a concentration-dependent manner. CONCLUSION: Osthole can inhibit the proliferation, collagen synthesis and phenotypic differentiation of lung fibroblast induced by TGFβ-1 in a concentration-dependent manner. The possible mechanism is related to the inhibition of Rac-1 expression and reactive oxygen species (ROS) production.