金雀异黄素对肺泡ⅱ型上皮细胞发生上皮-间质转化的影响.pdf

2016 年10月第23卷第10期 中国中医药信息杂志 ·63· 金雀异黄素对肺泡Ⅱ型上皮细胞发生上皮-间质转化的影响 王玲，刘辉国 武汉大学医院，湖北 武汉 430071 摘要：目的 观察金雀异黄素对转化生长因子-β1 （TGF-β1 ）诱导人肺腺癌A549 细胞发生上皮-间质转化 （EMT ）的影响，探讨其改善肺纤维化的作用机制。方法 CCK-8 法检测不同浓度 GEN 和/或 TGF-β1 干预细 胞 72 h 后的活性；RT-PCR 检测上皮及间质标记物的转变以及在转变过程中发挥关键调节作用的转录因子； Western blot 检测 p38 及 JNK 信号通路的变化。结果 TGF-β1 连续刺激 A549 细胞 72 h 可诱导其转化为成纤 维细胞表型，表现出更多间质标记物升高，内皮标记物降低；RT-PCR 检测结果显示，金雀异黄素可呈浓度依 赖性逆转这种现象；Western blot 检测结果显示，金雀异黄素与 TGF-β1 协同刺激 A549 细胞 24 h 可显著抑制 TGF-β1 诱导 p38 及 JNK 信号通路的磷酸化水平。结论 金雀异黄素可抑制 TGF-β1 诱导 A549 细胞发生 EMT 而发挥抗纤维化的作用。 关键词：金雀异黄素；上皮-间质转化；A549；肺纤维化；转化生长因子-β1 DOI：10.3969/j.issn.1005-5304.2016.10.015 中图分类号：R285.5 文献标识码：A 文章编号：1005-5304(2016)10-0063-04 Effects of Genistein on Epithelium-Mesenchyme Transition of Alveolar Type Ⅱ Cells WANG Ling, LIU Hui-guo (The Hospital of Wuhan University, Wuhan 430071, China) Abstract: Objective To investigate the effects of genistein on the epithelium-mesenchyme transition of alveolar type II cells; To discuss its mechanism of improving pulmonary fibrosis. Methods The activity of genistein with different concentrations and TGF-β1 intervention cells after 72 h were determined by CCK-8 method; epithelial and mesenchymal markers as well as the key regulatory factors in the transformation process were determined by RT-PCR; changes of p38 and JNK signaling pathways were determined by Western blot. Results The A549 cells were transformed into fibroblast phenotype stimulated by TGF-β1 for 72 h; mesenchymal markers increased and endothelial marker decreased. The results of RT-PCR indicated that genistein inhibited this phenomenon in a concentration-dependent manner. The resul