中华中医药学刊

补肺汤对慢性阻塞性肺疾病肺气虚证模型大鼠PINK1/Parkin信号通路介导的线粒体自噬的影响

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褚璨灿 ¹^,² , 陈光磊 ¹ , 郑宇皓 ¹ , 杨长福 ¹ , 朱星 ¹

中华中医药学刊 | 国家项目点击 2026,44(4): 110-116

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中华中医药学刊 |国家项目点击 2026 , 44 (4) : 110 -116

补肺汤对慢性阻塞性肺疾病肺气虚证模型大鼠PINK1/Parkin信号通路介导的线粒体自噬的影响

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褚璨灿¹^,², 陈光磊¹, 郑宇皓¹, 杨长福¹, 朱星¹

作者信息

^1.贵州中医药大学，贵州贵阳 550025

^2.安顺市人民医院，贵州安顺 561000

通讯作者:

朱星（1975-），男（土家族），贵州思南人，教授，博士研究生导师，博士，研究方向：中医防治肺系疾病基础。E-mail: 11863362@qq.com。

作者简介:

褚璨灿（1993-），女，贵州普定人，主治医师，博士在读，研究方向：中医防治肺系疾病基础。

Effect of Bufei Decoction（补肺汤）on Mitochondrial Autophagy Mediated by PINK1/Parkin Signaling Pathway in COPD Lung Qi Deficiency Syndrome Model Rats

Cancan CHU¹^,², Guanglei CHEN¹, Yuhao ZHENG¹, Changfu YANG¹, Xing ZHU¹

Affiliations

^1.Guizhou University of Traditional Chinese Medicine, Guiyang 550025, Guizhou, China

^2.People＇s Hospital of Anshun, Anshun 561000, Guizhou, China

出版时间: 2026-04-10 doi: 10.13193/j.issn.1673-7717.2026.04.024

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摘要

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目的

探讨补肺汤通过调控张力素同源物诱导的激酶蛋白1（phosphatase and tensin homolog-induced putative kinase protein 1，PINK1）/RBR结构域的E3泛素连接酶（parkin RBR E3 ubiquitin protein ligase，Parkin）信号通路介导的线粒体自噬水平干预慢性阻塞性肺疾病（chronic obstructive pulmonary disease，COPD）肺气虚证的作用机制。

方法

将大鼠随机分为对照组和造模组，采用被动吸入烟雾联合气道内滴注脂多糖的方法制造COPD肺气虚证大鼠模型。造模结束后，将造模组大鼠随机分为模型组、补肺汤中剂量组、补肺汤高剂量组、3-甲基腺嘌呤（3-methyladenine，3-MA）组。补肺汤中、高剂量组给予补肺汤药液灌胃（7.29、14.58 g·kg^-1），3-MA组以3-MA（15 mg·kg^-1）腹腔注射给药，每组每日药物干预1次，连续干预28 d。苏木素-伊红（hematoxylin-eosin，HE）染色观测肺组织病理改变；透射电镜下观测肺组织中肺泡Ⅱ型上皮细胞超微结构的变化；免疫组化（immunohistochemistry，IHC）检测肺组织中PINK1、Parkin、自噬受体蛋白P62（sequestosome 1，P62/SQSTM1）、微管相关蛋白1轻链3（microtubule-associated protein 1 light chain 3，LC3）蛋白的阳性表达；蛋白免疫印迹法（Western blot）检测肺组织线粒体中PINK1、Parkin、P62、LC3Ⅱ/LC3I蛋白表达；流式细胞术检测肺组织中线粒体膜电位变化情况；活性氧荧光探针（dichloro-dihydro-fluorescein diacetate，DCFH-DA）、线粒体超氧化物（mitochondrial superoxide，MitoSOX）红色荧光探针经流式细胞仪分别测定肺组织内活性氧（reactive oxygen species，ROS）、线粒体活性氧（mitochondrial reactive oxygen species，mtROS）水平；Western blot检测肺组织中核转录因子-κB P65（nuclear factor-κB P65，NF-κB P65）、磷酸化核转录因子-κB P65（phosphorylation of nuclear factor-κB P65，p-NF-κB P65）蛋白的表达。

结果

与对照组相比，模型组的肺组织损伤严重；与模型组相比，补肺汤中、高剂量组和3-MA组的病理损伤均有不同程度改善。与对照组相比，模型组肺组织中肺泡Ⅱ型上皮细胞及其线粒体结构损伤严重，线粒体减少，自噬小体增多；与模型组相比，补肺汤中、高剂量组和3-MA组肺泡Ⅱ型上皮细胞及其线粒体结构损伤程度较轻，线粒体相对增多，自噬小体减少。与对照组相比，模型组肺组织及肺组织线粒体中的PINK1、Parkin、LC3、LC3Ⅱ/LC3I的蛋白表达上调（P<0.01）、P62蛋白表达下调（P<0.01）；与模型组相比，补肺汤中、高剂量组和3-MA组肺组织及肺组织线粒体中的PINK1、Parkin、LC3、LC3Ⅱ/LC3I的蛋白表达下调（P<0.05，P<0.01）、P62蛋白表达上调（P<0.01）。与对照组相比，模型组肺组织的线粒体膜电位降低（P<0.01）、ROS和mtROS水平均上调（P<0.01）、NF-κB P65蛋白表达差异无统计学意义、p-NF-κB P65蛋白表达上调（P<0.01）；与模型组相比，补肺汤中、高剂量组和3-MA组肺组织的线粒体膜电位升高（P<0.01）、ROS和mtROS水平均下调（P<0.01）、NF-κB P65蛋白表达差异无统计学意义、p-NF-κB P65蛋白表达下调（P<0.01）。

结论

补肺汤能抑制肺组织ROS水平以及NF-κB P65的激活，从而减轻COPD肺气虚证肺部氧化损伤和炎症反应，其作用机制可能与抑制PINK1/Parkin信号通路介导的线粒体自噬水平有关。

关键词

补肺汤 / 慢性阻塞性肺疾病 / 线粒体自噬 / PINK1/Parkin信号通路 / 活性氧 / 核转录因子-κB

Abstract

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Objective

To investigate the mechanism of action of Bufei Decoction（补肺汤）in intervening chronic obstructive pulmonary disease（COPD）lung Qi deficiency by modulating the levels of mitochondrial autophagy mediated by phosphatase and tensin homolog-induced putative kinase protein 1（PINK1）/RBR domain E3 ubiquitin protein ligase（Parkin）signaling path-way.

Methods

The rats were randomly divided into the control group and the modeling group, and a rat model of COPD lung Qi deficiency was created by using passive smoke inhalation combined with intra-airway lipopolysaccharide drip. At the end of modeling, the rats in the modeling group were randomly divided into the model group, Bufei Decoction medium-dose group, Bufei Decoction high-dose group and 3-methyladenine（3-MA）group. The Bufei Decoction medium - and high-dose groups were given Bufei Decoction drug solution by gavage（7.29, 14.58 g·kg^-1）, and 3-MA group was given 3-MA（15 mg·kg^-1）by intraperitoneal injection, and each group was intervened with the drug once daily for 28 d consecutively. Hematoxylineosin（HE）staining was used to observe the pathological changes in lung tissues. The transmission electron microscopy was used to observe ultrastructural changes in alveolar type Ⅱ epithelial cells in lung tissues. The immunohistochemistry（IHC）method was used to detect the positive expressions of PINK1, Parkin, autophagy receptor protein P62（sequestosome 1, P62/SQSTM1）and microtubule-associated protein 1 light chain 3（LC3）proteins in lung tissues and protein immunoblotting（Western blot）to detect the expressions of PINK1 in mitochondria in lung tissues, Parkin, P62, LC3Ⅱ/LC3I protein expression in mitochondria of lung tissues. Flow cytometry detected the changes of mitochondrial membrane potential in lung tissues. The reactive oxygen species fluorescent probe dichloro-dihydro-fluorescein diacetate（DCFH-DA）and mitochondrial superoxide（MitoSOX）red fluorescent probe were used to determine the levels of reactive oxygen species（ROS）and mitochondrial reactive oxygen species（mtROS）in lung tissues by flow cytometry, respectively. Western blot detected the expressions of nuclear factor-κB P65（NF-κB P65）and phosphorylation of nuclear factor-κB P65（p-NF-κB P65）proteins in lung tissues.

Results

Compared with that in the control group, the lung tissue damage in the model group was severe. Compared with that in the model group, the pathological damages in the Bufei Decoction medium - and high-dose groups and 3-MA group were improved to different degrees. Compared with those of the control group, the lung tissue of the model group showed severe damage to alveolar type Ⅱ epithelial cells and their mitochondrial structure, with a decrease in mitochondria and an increase in autophagic vesicles. Compared with those of the model group, the alveolar type Ⅱ epithelial cells and their mitochondrial structure of the model group showed a lesser degree of damage in the Bufei Decoction medium - and high-dose groups and the 3-MA group, with a relatively higher increase in mitochondria and a decrease in autophagic vesicles. Compared with those in the control group, the protein expressions of PINK1, Parkin, LC3 and LC3Ⅱ/LC3I were up-regulated（P<0.01）and the protein expression of P62 was down-regulated（P<0.01）in the lung tissues and mitochondria of lung tissue in the model group. Compared with those in the model group, the protein expressions of PINK1, Parkin and LC3Ⅱ/LC3I were up-regulated（P<0.01）and the protein expression of P62 was down-regulated（P<0.01）in the lung tissues and mitochondria of lung tissue in the Bufei Decoction medium - and high-dose groups and 3-MA group. The protein expressions of PINK1, Parkin, LC3 and LC3Ⅱ/LC3I were down-regulated（P<0.05, P<0.01）and P62 protein expression was up-regulated（P<0.01）. Compared with those in the control group, the mitochondrial membrane potential of lung tissues in the model group was decreased（P<0.01）, the levels of ROS and mtROS were up-regulated（P<0.01）, the difference in the expression of NF-κB P65 protein was not statistically significant, and the expression of p-NF-κB P65 protein was up-regulated（P<0.01）. Compared with those of the model group, the mitochondrial membrane potential in the lung tissues of the Bufei Decoction medium - and high-dose groups and the 3-MA group were elevated（P<0.01）, the levels of ROS and mtROS were down-regulated（P<0.01）, and the differences in the expression of NF-κB P65 proteins were not statistically significant, and the expression of p-NF-κB P65 protein was down-regulated（P<0.01）.

Conclusion

Bufei Decoction inhibited lung tissue ROS levels and the activation of NF-κB P65, thereby attenuating the oxidative damage and inflammatory responses in the lung of COPD lung Qi deficiency, and its mechanism of action may be related to the inhibition of the level of mitochondrial autophagy mediated by the PINK1/Parkin signaling pathway.

Key words

Bufei Decoction（补肺汤） / chronic obstructive pulmonary disease / mitochondrial autophagy / PINK1/Parkin signaling pathway / reactive oxygen species / nuclear transcription factor-κB

引用本文

褚璨灿, 陈光磊, 郑宇皓, 杨长福, 朱星. 补肺汤对慢性阻塞性肺疾病肺气虚证模型大鼠PINK1/Parkin信号通路介导的线粒体自噬的影响. 中华中医药学刊, 2026 , 44 (4) : 110 -116 . DOI: 10.13193/j.issn.1673-7717.2026.04.024

Cancan CHU, Guanglei CHEN, Yuhao ZHENG, Changfu YANG, Xing ZHU. Effect of Bufei Decoction（补肺汤）on Mitochondrial Autophagy Mediated by PINK1/Parkin Signaling Pathway in COPD Lung Qi Deficiency Syndrome Model Rats[J]. Chinese Archives of Traditional Chinese Medicine, 2026 , 44 (4) : 110 -116 . DOI: 10.13193/j.issn.1673-7717.2026.04.024

基金

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国家自然科学基金项目(81960830)
贵州中医药大学-贵中医创新团队项目(贵中医TD合字〔2023〕002号)

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2026年第44卷第4期

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doi: 10.13193/j.issn.1673-7717.2026.04.024

首发时间：2026-10-14
出版时间：2026-04-10

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基金

国家自然科学基金项目(81960830)

贵州中医药大学-贵中医创新团队项目(贵中医TD合字〔2023〕002号)

作者信息

^1.贵州中医药大学，贵州贵阳 550025

^2.安顺市人民医院，贵州安顺 561000

通讯作者:

朱星（1975-），男（土家族），贵州思南人，教授，博士研究生导师，博士，研究方向：中医防治肺系疾病基础。E-mail: 11863362@qq.com。

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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