培土生金法对COPD小鼠骨骼肌线粒体能量代谢保护作用机制研究

Study on the protective effect and mechanism of the method of strengthening the spleen to benefit the lung on the mitochondrial energy metabolism of skeletal muscle of mice with chronic obstructive pulmonary disease

关键词:  培土生金法  参苓白术散  慢性阻塞性肺疾病  骨骼肌线粒体能量代谢

Key words:  method of strengthening the spleen to benefit the lung  Shenling Baizhu Powder  COPD  skeletal muscle mitochondrial energy metabolism

摘要:

[目的] 探讨培土生金法对慢性阻塞性肺疾病（COPD）小鼠骨骼肌线粒体能量代谢的保护作用及机制。[方法] 将50只小鼠随机分为5组，分别为对照组、模型组、参苓白术散高、中、低剂量组。采用烟熏方法（持续8周）复制COPD小鼠模型，熏烟4周后给予参苓白术散灌胃，实验结束后取腓肠肌组织并检测三磷酸腺苷（ATP）、二磷酸腺苷（ADP）及一磷酸腺苷（AMP）含量；Western blot法半定量分析腺苷酸活化蛋白激酶（AMPK）、p-AMPK、过氧化物酶体增殖活化受体γ共激活因子-1α（PGC-1α）、线粒体融合蛋白2（Mfn2）表达变化。[结果] 与对照组相比，模型组ATP/AMP显著降低（P<0.05）；与模型组相比，参苓白术散高剂量组ATP/AMP比值升高（P<0.05）。与对照组相比，模型组p-AMPK/AMPK比值降低，PGC-1α及Mfn2蛋白表达减少（P<0.05），与模型组相比，高、中剂量组p-AMPK/AMPK比值升高，高剂量组PGC-1α及Mfn2蛋白表达增加（P<0.05）。[结论] 培土生金法可通过提高p-AMPK、PGC-1α及Mfn2的表达以保护COPD小鼠线粒体能量代谢，从而维持COPD小鼠骨骼肌正常功能。

Abstract:

[Objective] To investigate the protective effect and mechanism of the method of strengthening the spleen to benefit the lung on the mitochondrial energy metabolism of skeletal muscle in chronic obstructive pulmonary disease (COPD) mice.[Methods] Fifty mice were randomly divided into 5 groups:control group,model group,Shenling Baizhu Powder high,medium and low dose groups. All mice except the control group were exposed to cigarette smoking for 8 weeks. After 4 weeks for exposure,Shenling Baizhu Powder was given by intragastric administration. After the experiment,the gastrocnemius tissue was taken and the contents of ATP,ADP and AMP were detected. The expression changes of AMPK,p-AMPK,PGC-1α and Mfn2 protein were analyzed by Western blot.[Results] Compared with the control group,the ATP/AMP in the model group decreased significantly (P<0.05);Compared with the model group,the ATP/AMP ratio in the high-dose group of Shenling Baizhu Powder increased (P<0.05). Compared with the control group,the level of p-AMPK/AMPK and PGC-1α and Mfn2 protein expression in the model group decreased (P<0.05). Compared with the model group,the p-AMPK/AMPK ratio increased in the high and medium dose Shenling Baizhu Powder groups increased,and the PGC-1α and Mfn2 protein expression in the high-dose group increased (P<0.05).[Conclusion] The method of strengthening the spleen to benefit the lung may increase the expression of p-AMPK,PGC-1α and Mfn2 to protect the energy synthesis of mitochondria in COPD mice,thus maintaining the skeletal muscle function of COPD mice.