补肺汤对肺纤维化小鼠肺组织中自噬相关蛋白的影响

Effect of Bufei Tang on the Autophagy Related Protein of Smad3,ATG5,p62 Involved in Pulmonary Fibrosis in Mouses with Pulmonary Fibrosis

目的:探讨补肺汤对肺纤维化中自噬相关蛋白Smad3、ATG5、p62的影响。方法:C57小鼠分为假手术组,模型组,补肺汤高、中、低剂量组,地塞米松组及补肺汤+地塞米松联合用药组。假手术组气管内滴注生理盐水,其余各组气管内滴注博莱霉素注射液。术后2 d给药,于第56天取肺组织分别进行HE染色、Mssson染色,并以免疫组化法及免疫蛋白印记法检测肺组织Smad3、ATG5、p62蛋白表达变化。结果:与模型组比较,补肺汤高剂量组、地塞米松组和联合用药组肺组织病理学显著改善,补肺汤高剂量组和联合用药组肺组织胶原活性显著降低;免疫组化检测显示补肺汤低剂量组Smad3表达减弱,补肺汤高剂量组及地塞米松组ATG5表达显著增强,补肺汤高、中剂量组和联合用药组p62表达显著增强;免疫蛋白印迹法显示各给药组Smad3、ATG5、p62蛋白表达差异与免疫组化结果基本一致。结论:补肺汤对肺纤维化组织病理学和胶原活性有明显影响,并对自噬相关蛋白Smad3、ATG5、p62有调节作用。

Objective：To investigate the effect of Bufei Tang on the autophagy related protein of Smad3, ATG5, p62 involved in pulmonary fibrosis in mice. Methods： C57 mice were divided into seven groups including sham group, model group, high dose group, middle dose group and low dose group of Bufei Tang, dexamethasone group, and mixed Bufei Tang with dexamethasone therapy group. Sham group was in intratracheal instillation of normal saline while the other groups with intratracheal instillation of bleomycin in- jection. To medicate each group in second day after postoperative. In the 56th days ,samples were treated with HE staining and Masson staining. Immunohistochemistry method was used to observe and record the lung tissue Smad3, ATG5, p62, and the variation protein ex- pression tendency of lung tissue Smad3, ATG5, p62 was analyzed by Western blotting. Results：The pathology of lung in high dose group of Bufei Tang,dexamethasone dose group and drug combination group had significantly improved compared with the model group. The lung tissue collagen activity declined in high dose group of Bufei Tang and drug combination group. Immunohistochemisty results showed that the expression of Smad3 in the high dose group was decreased, the expression of ATG5 in high dose group of Bufei Tang and dexa- methasone group was significantly increased, and the expression of p62 in the high dose group and middle dose group was significantly increased. Western-blotting results showed that the variation protein expression of Smad3, ATG5,p62 had the same results with immuno- hischemistry method. Conclusion：The Bufei Tang has significant effect on the pathological changes of lung fibrosis and the activity of collagen, and it has a regulatory effect on the changes of autophagy related protein Smad3, ATG5, p62.