12种中药醇提取物对TGF-β1诱导的A549细胞上皮间质转化的干预作用

Effects of 12 kinds of ethanol extracts of Chinese traditional medicine on TGF-β1 induced epithelial-mesenchymal transition in A549 cells

目的对茯苓、蒲黄、白及、鸡血藤、郁金、红花、牛膝、何首乌、黄芪、仙茅、鹿衔草、白化参共12种中药的醇提取物进行筛选,探讨12种中药醇提取物对转化生长因子-β1(TGF-β1)诱导的非小细胞肺癌A549细胞上皮间质转化(EMT)的干预作用。方法10 ng/ml TGF-β1诱导A549细胞;采用MTT法检测12种中药醇提取物对A549细胞增殖的影响;观察醇提取物对TGF-β1诱导的A549细胞形态变化的影响;采用Western blot方法检测醇提取物对TGF-β1诱导的EMT相关标志蛋白的表达,以及对EMT相关信号转导的影响。结果 10 ng/ml的TGF-β1诱导A549细胞48 h后使其发生EMT过程;何首乌醇提取物不影响A549细胞增殖;与TGF-β1组相比较,100μg/ml的何首乌醇提取物能逆转A549细胞形态变化过程;与TGF-β1组相比较,100μg/ml的何首乌醇提取物降低vimentin蛋白表达水平(P<0.05),上调E-cadherin蛋白表达水平(P<0.05);与TGF-β1组比较,100μg/ml的何首乌醇提取物降低pSmad2和p-Erk蛋白表达水平(P<0.05)。结论何首乌醇提取物具有抑制TGF-β1诱导的A549细胞上皮间质转化过程的作用,这可能与Smad2和Erk1/2信号通路有关。

Objective To investigate the intervention effects of 12 kinds of ethanol extracts of Chinese traditional medicine on TGF-β1 induced epithelialmesenchymal transition( EMT) in non-small cell lung cancer A549 cells,including Fuling( Poria),Puhuang( Typhae Pollen),Baiji( Bletillae Rhizoma),Jixueteng( Spatholobi Caulis),Yujin( Curcumae Radix),Honghua( Carthami Flos),Niuxi( Achyranthis Bidentatae Radix),Heshouwu( Polygoni multiflora Radix),Huangqi( Astragali seu Hedysari Radix),Xianmao( Curculigins Rhizoma),Luxiancao( Pyrolae Herba) and Baihuashen. Methods The A549 cells were stimulated by 10 ng/ml TGF-β1. The effects of 12 kinds of ethanol extracts of Chinese traditional medicine on the proliferation of A549 cells were evaluated by MTT assay. The effects of ethanol extracts on the changes of cell morphology induced by TGF-β1 were observed. The effects of ethanol extracts on the expressions of EMT associated protein biomarkers and the activity of EMT associated signal transduction were detected by Western blot. Results After stimulation with 10 ng/ml TGF-β1 for 48 hours,the A549 cells showed the EMT process. The ethanol extract of Heshouwu showed no effect on the proliferation of A549 cells. Compared with the TGF-β1 group,the ethanol extract of Heshouwu at 100 μg/ml could reverse the morphological changes of A549 cells. Compared with the TGF-β1 group,the ethanol extract of Heshouwu at 100 μg/ml could decrease the protein expression level of vimentin( P<0.05) and increase the protein expression level of E-cadherin( P<0.05). Compared with the TGF-β1 group,the ethanol extract of Heshouwu at 100 μg/ml could decrease the protein expression levels of p-Smad2 and p-Erk( P<0.05). Conclusion The ethanol extract of Heshouwu can inhibit the TGF-β1 induced epithelial-mesenchymal transition in A549 cells,which may be associated with Smad2 and Erk1/2 signaling pathway.