金水缓纤组分方Ⅱ调控PI3K/Akt通路抑制TGF-β1诱导的人胚肺成纤维细胞活化

Inhibition of Jinshui Huanxian FormulaⅡon TGF-β1-Induced Activation of Human Embryonic Lung Fibroblasts by PI3K/Akt Pathway

目的:探讨金水缓纤组分方Ⅱ调控PI3K/Akt通路影响成纤维细胞活化、增殖及迁移干预特发性肺纤维化(IPF)的机制。方法:以TGF-β1作用MRC-5细胞诱导其活化,采用MTT法检测细胞活力筛选药物浓度;Western Blot检测低(30.63μg/mL)、中(61.25μg/mL)、高(91.88μg/mL)浓度金水缓纤组分方Ⅱ对活化MRC-5细胞CollagenⅠ、N-cadherin、α-SMA、PCNA、p-PI3K、p-Akt蛋白表达的影响;免疫荧光技术检测各组细胞CollagenⅠ、α-SMA表达;EdU法和划痕法检测金水缓纤组分方Ⅱ对活化MRC-5细胞增殖及迁移能力的影响。加入bpv(HOpic)激活PI3K/Akt通路和特异性抑制剂LY294002考察金水缓纤组分方Ⅱ调控PI3K/Akt通路对成纤维细胞活化、增殖及迁移的影响。结果:中、高浓度金水缓纤组分方Ⅱ均可抑制CollagenⅠ、N-cadherin、α-SMA、PCNA、p-PI3K、p-Akt表达,抑制MRC-5细胞的增殖和迁移能力;LY294002可增强金水缓纤组分方Ⅱ对MRC-5细胞活化、增殖和迁移的影响,bpv可抑制金水缓纤组分方Ⅱ的影响。结论:金水缓纤组分方Ⅱ调控PI3K/Akt通路抑制TGF-β1诱导的MRC-5细胞的增殖及迁移能力。

Objective:To investigate the mechanism of Jinshui huanxian formulaⅡintervening idiopathic pulmonary fibrosis(IPF)by regulating PI3K/Akt pathway and affecting fibroblast activation,proliferation and migration.Methods:TGF-β1 was applied to MRC-5 cells to induce the activation Cell viability was detected by MTT assay to screen the drug concentration.The effect of low(30.63μg/mL),medium(61.25μg/mL)and high(91.88μg/mL)concentrations of Jinshui huanxian formulaⅡon the protein expressions of CollagenⅠ,N-cadherin,α-SMA,PCNA,p-PI3K,p-Akt in activated MRC-5 cells was detected by Western Blot.CollagenⅠandα-SMA expressions ineachgroup were detected by immunofluorescence.EdU method and scratch method were used to detect the effect of Jinshui huanxian formulaⅡon activated MRC-5 cell proliferation and migration.The PI3K/Akt pathway was activated by adding bpv(HOpic)and the specific inhibitor LY294002 was used to study the effects of the regulation of PI3K/Akt pathway by Jinshui huanxian formulaⅡon the activation,proliferation and migration of fibroblasts.Results:The expressions of CollagenⅠ,N-cadherin,α-SMA,PCNA,p-PI3K and p-AKT in medium and high concentrations of Jinshui huanxian formulaⅡgroups were inhibited,and the proliferation and migration of MRC-5 cells were inhibited.LY294002 could enhance the effect of Jinshui huanxian formulaⅡon the activation,proliferation and migration of MRC-5 cells,and bpv could inhibit the effect of Jinshui huanxian formulaⅡ.Conclusion:Jinshui huanxian formulaⅡcan significantly inhibit TGF-β1-induced fibroblast proliferation and migration of MRC-5 cells by regulating PI3K/Akt pathway.