黄芪注射液通过抑制EMT过程中pERK1/2信号通路减轻肺纤维化

Astragalus injection attenuates pulmonary fibrosis by inhibiting pERK1/2 signal pathway in process of EMT

目的:研究黄芪注射液对肺成纤维细胞上皮-间充质转化(EMT)过程中pERK1/2信号通路的影响,探讨黄芪注射液抑制肺纤维化的作用机制。方法:体外正常培养小鼠肺成纤维细胞(L-929)并分为5组:正常对照组(L-929+DMEM完全培养基)、模型组(L-929+5 ng/ml TGF-β1诱导)、治疗低剂量组(L-929+5 ng/ml TGF-β1+100 mg/ml黄芪注射液)、治疗中剂量组(L-929+5 ng/ml TGF-β1+200 mg/ml黄芪注射液)、治疗高剂量组(L-929+5 ng/ml TGF-β1+300 mg/ml黄芪注射液),孵育24 h,倒置相差显微镜观察细胞形态变化,RT-PCR检测α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原(CollagenⅠ)基因表达,Western blot检测细胞外信号调节激酶1/2(ERK1/2)磷酸化蛋白表达。结果:与对照组相比,模型组α-SMA、CollagenⅠmRNA表达、pERK1/2蛋白表达升高(P<0.05);与模型组相比,治疗低、中、高剂量组α-SMA、CollagenⅠmRNA表达、pERK1/2蛋白表达降低(P<0.05),且呈剂量依赖性。结论:黄芪注射液可能通过抑制EMT过程中的pERK1/2信号通路减轻肺纤维化。

Objective:To study effect of astragalus injection on pERK1/2 signal pathway in process of epithelial-mesenchymal transformation(EMT)of pulmonary fibroblasts,and to explore mechanism of astragalus injection in inhibiting pulmonary fibrosis.Methods:Mice lung fibroblasts(L-929)were cultured in vitro and divided into 5 groups:normal control group(L-929+DMEM complete medium),model group(L-929+5 ng/ml TGF-β1 stimulation),treatment low-dose group(L-929+5 ng/ml TGF-β1+100 mg/ml astragalus injection),treatment middle-dose group(L-929+5 ng/ml TGF-β1+200 mg/ml astragalus injection),treatment high-dose group(L-929+5 ng/ml TGF-β1+300 mg/ml astragalus injection),and incubated for 24 h.Morphological changes of cells were observed under inverted phase contrast microscope,gene expressions ofα-smooth muscle actin(α-SMA)and CollagenⅠwere detected by RT-PCR.Western blot was used to detect phosphorylated extracellular signal-regulated kinase 1 and 2(ERK1/2)proteins expressions.Results:Compared with control group,expressions ofα-SMA and CollagenⅠmRNA,pERK1/2 protein in model group were increased(P<0.05).Compared with model group,expressions ofα-SMA and CollagenⅠmRNA and pERK1/2 protein in low,middle and high doses groups were decreased(P<0.05),which showed a dose-dependent manner.Conclusion:Astragalus injection may alleviate pulmonary fibrosis by inhibiting pERK1/2 signal pathway in process of EMT.