金匮肾气丸通过AMPK/mTOR途径抑制肺组织细胞凋亡和自噬在大鼠慢性阻塞性肺疾病中的作用

Effect of Jingui Shenqi Pill on inhibition of apoptosis and autophagy of lung tissue through AMPK/mTOR pathway in rats with chronic obstructive pulmonary disease

关键词:  金匮肾气丸  慢性阻塞性肺疾病  细胞凋亡  自噬  AMPK/mTOR途径

Key words:  Jingui Shenqi Pill  chronic obstructive pulmonary disease  apoptosis  autophagy  AMPK/mTOR pathway

摘要:

[目的] 探究金匮肾气丸对慢性阻塞性肺疾病（COPD）的作用及其可能的作用机制。[方法] 40只SD大鼠随机分为空白对照组、COPD组、金匮肾气丸3.7 g/kg组、金匮肾气丸7.4 g/kg组和地塞米松组，每组各8只。除空白对照组外，其余组大鼠采用烟熏联合气管内滴注脂多糖（LPS）法建立COPD大鼠模型。各组给予相应药处理AniRes2005动物肺功能分析系统检测大鼠肺功能；酶联免疫吸附（ELISA）试剂盒检测白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和白细胞介素-17（IL-17）水平；苏木精-伊红（HE）染色检测肺组织病理学变化；TUNEL染色检测肺组织细胞凋亡；蛋白质免疫印迹（Western blot）检测自噬及磷酸腺苷活化蛋白激酶（AMPK）/哺乳动物雷帕霉素靶蛋白（mTOR）途径相关蛋白的表达。[结果] 与空白对照组相比，COPD组大鼠肺功能降低，IL-1β、TNF-α、IL-6和IL-17水平明显升高（P<0.05），炎症评分、细胞凋亡率和LC3Ⅱ/LC3Ⅰ、Beclin1、p-AMPK/AMPK蛋白表达水平明显升高（P<0.05），p62和p-mTOR/mTOR蛋白表达水平明显降低（P<0.05）；与COPD组相比，金匮肾气丸3.7 g/kg组、金匮肾气丸7.4 g/kg组和地塞米松组大鼠肺功能升高，IL-1β、TNF-α、IL-6和IL-17水平明显降低（P<0.05），炎症评分、细胞凋亡率和LC3Ⅱ/LC3Ⅰ、Beclin1、p-AMPK/AMPK蛋白表达水平明显降低（P<0.05），p62和p-mTOR/mTOR蛋白表达水平明显升高（P<0.05）。[结论] 金匮肾气丸可能通过AMPK/mTOR途径抑制肺组织细胞凋亡和自噬，延缓大鼠慢性阻塞性肺疾病进展。

Abstract:

[Objective] To explore the effect of Jingui Shenqi Pills on chronic obstructive pulmonary disease (COPD) and its possible mechanism.[Methods] Forty SD rats were randomly divided into control group,COPD group,Jingui Shenqi Pills 3.7 g/kg group,Jingui Shenqi Pills 7.4 g/kg group and dexamethasone group,8 rats in each group. Except for the control group,the rats in the remaining groups were established with COPD rats by smoking combined with intratracheal lipopolysaccharide (LPS) method. Each group was given corresponding drug treatment. AniRes2005 animal lung function analysis system was used to detect rat lung function. ELISA kits were used to detect the levels of IL-1β,TNF-α,IL-6 and IL-17. HE staining was used to detect lung histopathological changes. TUNEL staining was used to detect the cell apoptosis of lung tissue. Western blot was used to detect the autophagy and AMPK/mTOR pathway related protein expression.[Results] Compared with the control group,the lung function of rats in COPD group was reduced,and the levels of IL-1β,TNF-α,IL-6 and IL-17 were significantly increased (P<0.05),inflammation score,apoptosis rate and LC3Ⅱ/LC3Ⅰ,Beclin1,p-AMPK/AMPK protein expression levels were significantly increased (P<0.05),p62 and p-mTOR/mTOR protein expression levels were significantly reduced (P<0.05);compared with COPD group,pulmonary function was increased in the Jingui Shenqi Pills 3.7 g/kg group,Jingui Shenqi Pill 7.4g/kg group and the dexamethasone group,and the levels of IL-1β,TNF-α,IL-6 and IL-17 were significantly reduced (P<0.05),inflammation score,apoptosis rate and LC3Ⅱ/LC3Ⅰ,Beclin1,p-AMPK/AMPK protein expression levels were significantly reduced (P<0.05),p62 and p-mTOR/mTOR protein expression levels were significantly increased (P<0.05).[Conclusion] Jingui Shenqi Pills may inhibit the apoptosis and autophagy of lung tissue through the AMPK/mTOR pathway,and delay the progression of chronic obstructive pulmonary disease in rats.