参芪补中方通过激活AMPK/SIRT1/PGC-1α改善COPD肺脾气虚证大鼠线粒体功能障碍

doi:10.12122/j.issn.1673-4254.2025.05.09

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (5): 969-976.doi: 10.12122/j.issn.1673-4254.2025.05.09

参芪补中方通过激活AMPK/SIRT1/PGC-1α改善COPD肺脾气虚证大鼠线粒体功能障碍

张璐¹^,²(), 丁焕章³, 许浩燃¹^,², 陈珂¹^,², 许博文¹^,², 杨勤军², 吴迪¹, 童佳兵²^,⁴, 李泽庚¹^,⁴()

^1.安徽中医药大学第一附属医院，安徽合肥 230031
^2.安徽中医药大学，安徽合肥 230038
^3.阜阳师范大学第一附属医院，安徽阜阳 236037
^4.中医药防治肺系重大疾病应用转化安徽省重点实验室，安徽合肥 230031

收稿日期:2024-12-19 出版日期:2025-05-20 发布日期:2025-05-23
通讯作者: 李泽庚 E-mail:1195263773@qq.com;ahzyfb@sina.com
作者简介:张璐，在读博士研究生，E-mail: 1195263773@qq.com
基金资助:
国家自然科学基金联合重点项目(U20A20398);合肥综合性国家科学中心大健康研究院新安医学与中医药现代化研究所“揭榜挂帅”重大突破项目(2023CXMMTCM005)

Shenqi Buzhong Formula ameliorates mitochondrial dysfunction in a rat model of chronic obstructive pulmonary disease by activating the AMPK/SIRT1/PGC-1α pathway

Lu ZHANG¹^,²(), Huanzhang DING³, Haoran XU¹^,², Ke CHEN¹^,², Bowen XU¹^,², Qinjun YANG², Di WU¹, Jiabing TONG²^,⁴, Zegeng LI¹^,⁴()

^1.First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230031, China
^2.Anhui University of Chinese Medicine, Hefei 230038, China
^3.Affiliated First Hospital of Fuyang Normal University, Fuyang 236037, China
^4.Anhui Provincial Key Laboratory of Application and Transformation of Traditional Chinese Medicine in Prevention and Treatment of Major Pulmonary Diseases, Hefei 230031, China

Received:2024-12-19 Online:2025-05-20 Published:2025-05-23
Contact: Zegeng LI E-mail:1195263773@qq.com;ahzyfb@sina.com

摘要/Abstract

摘要：

目的基于AMP依赖的蛋白激酶（AMPK）/沉默调节蛋白1（SIRT1）/过氧化物酶体增殖物激活受体γ辅激活因子1α（PGC-1α）通路探讨参芪补中方对COPD大鼠线粒体功能障碍的影响。方法将60只SD大鼠随机分为正常组（Control）、模型组（Model）、参芪补中方低剂量组（SQBZ-L）、参芪补中方中剂量组（SQBZ-M）、参芪补中方高剂量组（SQBZ-H）、氨茶碱组（APL），10只/组。除正常组外，其余各组通过脂多糖气道滴注、香烟烟熏联合番泻叶浸液灌胃构建COPD模型，参芪补中方组和氨茶碱组分别于第50天进行参芪补中方和氨茶碱灌胃，2次/d，连续灌胃给药4周。检测各组大鼠肺功能，HE染色和透射电镜观察各组大鼠肺组织病理形态和超微结构改变，WST-1法、比色法、TBA法、JC-1法分别检测大鼠肺组织中超氧化物歧化酶（SOD）活性、三磷酸腺苷（ATP）、丙二醛（MDA）含量及线粒体膜电位，流式细胞术检测大鼠ROS平均荧光强度，Western blotting法检测肺组织中P-AMPKα、AMPKα、SIRTI、PGC-1α蛋白表达水平。结果与对照组相比，模型组大鼠肺功能下降、肺组织病理损伤严重（P<0.01），SOD活性、ATP水平及线粒体膜电位下降（P<0.01），MDA水平和ROS平均荧光强度显著提高（P<0.01），线粒体损伤加重，P-AMPKα、SIRTI、PGC-1α蛋白表达水平下降（P<0.01）；与模型组相比，参芪补中方各剂量组与氨茶碱组能有效改善COPD大鼠的肺功能、肺组织病理损伤（P<0.05，P<0.01），提高SOD活性、ATP含量及线粒体膜电位（P<0.05，P<0.01），降低MDA含量和ROS平均荧光强度（P<0.05，P<0.01），上调P-AMPKα、SIRTI、PGC-1α蛋白表达水平（P<0.05，P<0.01）；参芪补中方高剂量组与氨茶碱组相比较，差异无统计学意义（P>0.05）。结论参芪补中方可能通过激活AMPK/SIRT1/PGC-1α通路改善COPD大鼠的线粒体功能障碍。

关键词: 慢性阻塞性肺疾病, 参芪补中方, 线粒体功能障碍, 线粒体生物发生, AMPK/SIRT1/PGC-1α通路

Abstract:

Objective To explore the mechanism of Shenqi Buzhong (SQBZ) Formula for alleviating mitochondrial dysfunction in a rat model of chronic obstructive pulmonary disease (COPD) in light of the AMPK/SIRT1/PGC-1α pathway. Methods Fifty male SD rat models of COPD, established by intratracheal lipopolysaccharide (LPS) instillation, exposure to cigarette smoke, and gavage of Senna leaf infusion, were randomized into 5 groups (n=10) for treatment with saline (model group), SQBZ Formula at low, moderate and high doses (3.08, 6.16 and 12.32 g/kg, respectively), or aminophylline (0.024 g/kg) by gavage for 4 weeks, with another 10 untreated rats as the control group. Pulmonary function of the rats were tested, and pathologies and ultrastructural changes of the lung tissues were examined using HE staining and transmission electron microscopy. The levels of SOD, ATP, MDA, and mitochondrial membrane potential in the lungs were detected using WST-1, colorimetric assay, TBA, and JC-1 methods. Flow cytometry was used to analyze ROS level in the lung tissues, and the protein expression levels of P-AMPKα, AMPKα, SIRTI, and PGC-1α were detected using Western blotting. Results The rat models of COPD showed significantly decreased lung function, severe histopathological injuries of the lungs, decreased pulmonary levels of SOD activity, ATP and mitochondrial membrane potential, increased levels of MDA and ROS, and decreased pulmonary expressions of P-AMPKα, SIRTI, and PGC-1α proteins. All these changes were significantly alleviated by treatment with SQBZ Formula and aminophylline, and the efficacy was comparable between high-dose SQBZ Formula group and aminophylline group. Conclusion SQBZ Formula ameliorates mitochondrial dysfunction in COPD rats possibly by activating the AMPK/SIRT1/PGC-1α pathway.

Key words: chronic obstructive pulmonary disease, Shenqi Buzhong Formula, mitochondrial dysfunction, mitochondrial biogenesis, AMPK/SIRT1/PGC-1α pathway

张璐, 丁焕章, 许浩燃, 陈珂, 许博文, 杨勤军, 吴迪, 童佳兵, 李泽庚. 参芪补中方通过激活AMPK/SIRT1/PGC-1α改善COPD肺脾气虚证大鼠线粒体功能障碍[J]. 南方医科大学学报, 2025, 45(5): 969-976.

Lu ZHANG, Huanzhang DING, Haoran XU, Ke CHEN, Bowen XU, Qinjun YANG, Di WU, Jiabing TONG, Zegeng LI. Shenqi Buzhong Formula ameliorates mitochondrial dysfunction in a rat model of chronic obstructive pulmonary disease by activating the AMPK/SIRT1/PGC-1α pathway[J]. Journal of Southern Medical University, 2025, 45(5): 969-976.

图/表 7

图1 参芪补中方对大鼠进食量的影响

Fig.1 Effect of Shenqi Buzhong Formula on food intake in rats. **P<0.01 vs Control group; #P<0.05, ##P<0.01 vs Model group.

图2 参芪补中方对大鼠肺功能的影响

Fig.2 Effect of Shenqi Buzhong Formula on lung function in COPD rats. **P<0.01 vs Control group; #P<0.05, ##P<0.01 vs Model group.

图3 参芪补中方对大鼠肺组织病理形态的影响

Fig.3 Effect of Shenqi Buzhong Formula on histopathological injuries in the lung tissue of COPD rats (HE staining, original magnification: ×200). A: Control group; B: Model group; C: SQBZ-L group; D: SQBZ-M group; E: SQBZ-H group; F: APL group.

图6 参芪补中方对大鼠ATP含量的影响

Fig.6 Effects of Shenqi Buzhong Formula on ATP content in lung tissues of COPD rats. **P<0.01 vs Control group. #P<0.05, ##P<0.01 vs Model group.

图7 参芪补中方对大鼠MDA、SOD水平的影响

Fig.7 Effects of Shenqi Buzhong Formula on MDA and SOD levels in COPD rats. **P<0.01 vs Control group. #P<0.05, ##P<0.01 vs Model group.

图8 参芪补中方对大鼠肺泡Ⅱ型上皮细胞线粒体形态的影响

Fig.8 Effects of Shenqi Buzhong Formula on mitochondrial morphology of rat alveolar type II epithelial cells (×10000). A: Control group; B: Model group; C: SQBZ-L group; D: SQBZ-M group; E: SQBZ-H group; F: APL group.

图9 参芪补中方对COPD大鼠P-AMPKα、AMPKα、SIRTI、PGC-1α蛋白表达的影响

Fig.9 Effects of Shenqi Buzhong Formula on protein expressions of P-AMPKα, AMPKα, SIRTI, and PGC-1α in the lung tissues of COPD rats. **P<0.01 vs Control group. #P<0.05, ##P<0.01 vs Model group. A: Control; B: Model; C: SQBZ-L; D: SQBZ-M; E: SQBZ-H; F: APL.

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参芪补中方通过激活AMPK/SIRT1/PGC-1α改善COPD肺脾气虚证大鼠线粒体功能障碍

Shenqi Buzhong Formula ameliorates mitochondrial dysfunction in a rat model of chronic obstructive pulmonary disease by activating the AMPK/SIRT1/PGC-1α pathway

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