黄芪总黄酮对特发性肺纤维化miRNA-21、let-7 d及TGF-β/smad信号干预作用  

Relationship between miRNA-21,let-7 d and Autophagy in Idiopathic Pulmonary Fibrosis and The Intervention Effect of Astragalus Total Flavonoids

作　　者：徐昌君[1] 王鹏飞 黄雅薇 朱星[1] 陈云志[1] 欧江琴[1] 丁倩[1] 杨长福[1] 赵宗江[2] 张莉莉[3] XU Changjun;WANG Pengfei;HUANG Yawei;ZHU Xing;OU Jiangqin DING Qian;YANG Changfu;ZHAO Zongjiang;ZHANG Lili(Guiyang University of Chinese Medicine, Guizhou 550002, Guiyang, China;Beijing University of Chinese Medicine, Beijing 100026,China;Nursing School of Guizhou Provinical People＇ s Hospital, Guizhou 550002, Guiyang,)

机构地区：[1]贵阳中医学院,贵阳贵州550002 [2]北京中医药大学,北京101029 [3]贵州省人民医院护士学校,贵阳贵州550003

出　　处：《中华中医药学刊》2018年第6期1308-1311,I0006,I0007,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81660735);国家自然科学基金地区科学基金项目(81260587);贵州省科学技术基金项目(黔科合J字[2014]2034号);贵州省教育厅青年科技人才成长项目(黔教合KY字[2016]189)

摘　　要：目的：研究黄芪总黄酮对BLM诱导的肺纤维化模型miRNA-21、let-7 d表达水平和TGF-β1/smad信号干预作用。方法：C57BL/6小鼠随机分为假手术组、模型组、黄芪总黄酮组。模型组及药物处理组小鼠气管内一次性滴注博莱霉素（10 mg/kg）,假手术组予等量生理盐水,造模后治疗组予黄芪总黄酮（43 mg/kg）灌胃,假手术组和模型组与等量生理盐水灌胃。第28天取材,进行HE、Masson染色分析肺组织形态及胶原蛋白表达水平,羟脯氨酸含量测定胶原蛋白含量,realtime-PCR分析miRNA-21、let-7 d表达变化,Western-Bloting检测TGF-β/smad信号及EMT相关蛋白表达。结果：BLM诱导肺纤维化miRNA-21明显上调,而let-7 d下调,在此过程中信号蛋白smad7表达增强mad3磷酸化水平减弱,EMT标记蛋白α-SMA表达增高而E-cadherin减少,黄芪总黄酮对上述改变具有逆转作用,但对let-7 d表达无影响。结论：黄芪总黄酮可通过抑制miRNA-21,增加smad7表达,使TGF-β1/smad激活能力下降,抑制肺纤维化EMT作用。Objective： To explore the intervention effect of Astragalus total flavone on BLM induced pulmonary fibrosis model＇s expression levels of miRNA-21,let-7 d and TGF-β1/Smad intervention. Methods： C57 BL/6 mice were randomly divided into model group and flavone group which were given BLM by one-time intratracheal instillation,and control group which were given the same amount of saline for 28 days. HE and Masson staining analyzed the inflammation and collagen expression level and realtime PCR tested the miRNA-21 and let-7 d expressions. Western bloting detected autophagy proteins and TGF-beta/Smad signal related protein. Results： In the lung tissue induced by BLM,HE and Masson staining showed significant inflammatory response and collagen deposition,in which miRNA-21 was up-regulated and let-7 d down-regulated followed a lower autophagy activity and Smad signal activation. Astragalus total flavone decreased expression of miRNA-21 and significantly increased LC3 BⅡ,but without the significant change of let-7 d.At same time,the phosphorylate Smad3（ p-Smad3） was inhibited with Smad7 enhanced. Conclusion： In BLM induced pulmonary fibrosis,Astragalus total flavone can inhibit miRNA-21 and increase Smad7 to reduce the activity of TGF-β1/Smad and inhibit pulmonary fibrosis.

关 键 词：特发性肺纤维化 MIRNA-21 LET-7 d TGF-Β1/SMAD 黄芪总黄酮 

分 类 号：R285.5[医药卫生—中药学]