摘要
目的:研究黄芪甲苷对特发性肺纤维化自噬活性影响及其磷酯酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/mTOR信号的调控作用,探讨黄芪的抗肺纤维化机制。方法:c57bl/6小鼠分为假手术组、模型组、黄芪甲苷低、中、高剂量组、泼尼松组。博莱霉素(BLM)10 mg·kg-1诱导建立肺纤维化动物模型,以黄芪甲苷低、中、高剂量(25,50,100 mg·kg-1)干预,分别于3,7,14,28 d取材,进行苏木素-伊红(HE)染色观察肺组织形态变化,马松(Masson)三色染色、羟脯氨酸测定观察胶原表达水平,免疫组化及蛋白质免疫印迹(Western bolt)分析自噬标记蛋白和PI3K/Akt/mTOR信号蛋白表达水平。结果:与空白组比较,模型组肺组织炎症反应明显、胶原蛋白显著增加(P<0.01),PI3K/Akt/mTOR信号增强,自噬活性下降,与模型组比较,黄芪甲苷中、高剂量明显抑制肺组织炎症反应,下调转化生长因子-β1(TGF-β1)(P<0.01)和胶原蛋白(P<0.05)表达,提高LC3Ⅱ/Ⅰ和(beclin-1)表达(P<0.05),减少p62积累(P<0.05),同时抑制PI3K,Akt,mTOR磷酸化水平(P<0.05),减少ɑ-平滑肌动蛋白(ɑ-SMA)而提高E-钙黏素(E-cadherin)表达(P<0.01)。结论:黄芪甲苷通过抑制肺组织炎性反应、下调TGF-β1表达,抑制PI3K/Akt/mTOR信号增强肺组织细胞自噬活性,阻止上皮间质转分化(EMT)过程。
Objective： To study the regulatory effect of phosphatidylinasitol 3-kinase（ PI3K）/Akt/mTOR signal on the autophagy in idiopathic pulmonary fibrosis and the intervention effect of astragaloside.Method： The c57bl/6 Mice induced by bleomycin（ BLM） were given astragaloside with low（ 25 mg·kg-1）,medium（ 50 mg·kg-1） and high（ 100 mg·kg-1） dose by gavage. At 3,7,14,28 days,lung tissues were collected for the analysis by hematoxylin-eosin（ HE）,Masson,immunohistochemical,and Western blot. Result：Astragaloside with medium dose and high dose significantly inhibited the inflammatory reaction of lung tissues,down-regulated expression of transforming growth factor-β1（ TGF-β1）, increased LC3 Ⅱ/Ⅰ and beclin-1expressions,decreased p62,inhibited phosphorylation levels of PI3 K,Akt and mTOR,and strengthened cell autophagy activity. Conclusion： Astragaloside stopped the epithelial mesenchymal transdifferentiation（ EMT）process by inhibiting the inflammatory reaction of lung tissues and the expression of TGF-β1,strengthening the autophagy activity and inhibiting PI3K/Akt/mTOR signal.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2017年第18期75-82,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81660735
81260587)
贵州省教育厅青年科技人才成长项目(黔科合LH字[2016]189)
