吡非尼酮抑制TGF-β1诱导的人肺成纤维细胞表型转化∗

摘要

AIM:To investigate the effect of pirfenidone on transforming growth factor-β1 (TGF-β1)-induced fibroblast-to-myofibroblast transition in vitro. METHODS:The cell viability was measured by MTT assay. The prolifera-tion of human lung fibroblasts ( HLFs) was detected by EdU incorporation. Migratory and invasive abilities were measured by Boyden chamber assay. The α-smooth muscle actin (α-SMA) protein expression was determined by Western blot and immunofluorescence. The mRNA expression of α-SMA and type Ⅰ and Ⅲ collagens was evaluated by RT-qPCR. RE-SULTS:Pirfenidone at different concentrations (0. 1, 0. 2, 0. 3, 0. 5 and 0. 8 mg/L) had no cytotoxic effect on the HLFs, and pirfenidone at 0. 2 mg/L was used for the intervention. Pretreatment of the HLFs with 0. 2 mg/L pirfenidone prior to TGF-β1 not only markedly suppressed the changes of proliferation, migration, invasion and reorganization of actin cytoskeleton in the HLFs (P<0. 01), but also down-regulated the expression ofα-SMA and typeⅠandⅢcollagens trig-gered by TGF-β1 ( P<0. 05 ) . CONCLUSION: Pirfenidone has an inhibitory effect on TGF-β1-induced activated cell functions and fibroblast-to-myofibroblast transition in HLFs.%目的：：研究吡非尼酮(PFD)是否抑制转化生长因子β1(TGF-β1)诱导的人肺成纤维细胞(HLFs)表型转化。方法：MTT法检测细胞存活率；EdU法检测细胞的增殖能力；Transwell实验检测细胞的迁移和侵袭能力,Western blot法和细胞免疫荧光检测α-平滑肌肌动蛋白(α-SMA)的蛋白水平,实时荧光定量PCR检测α-SMA和Ⅰ、Ⅲ型胶原蛋白的mRNA表达水平。结果：不同浓度的吡非尼酮(0．1、0．2、0．3、0．5和0．8 mg/L)无明显的细胞毒性作用,后续实验应用0．2 mg/L为干预浓度。吡非尼酮(0．2 mg/L)预处理HLFs能明显地抑制TGF-β1诱导的细胞增殖、迁移和侵袭能力,下调Ⅰ、Ⅲ型胶原蛋白的mRNA表达水平( P<0．05),并且干扰TGF-β1诱导的细胞骨架重组和表型转化,使α-SMA的mRNA和蛋白水平均下降( P<0．05)。结论：吡非尼酮能有效地抑制TGF-β1所诱导的HLFs细胞功能和表型转化。