氢气水调控自噬对百草枯中毒大鼠肺纤维化的影响

刘永骏; 王颖; 杨眉; 殷娥高; 李婷; 董昭兴

doi:10.12259/j.issn.2095-610X.S20210918

氢气水调控自噬对百草枯中毒大鼠肺纤维化的影响

doi: 10.12259/j.issn.2095-610X.S20210918

刘永骏^{1, 2},
王颖¹,
杨眉¹,
殷娥高¹,
李婷³,
董昭兴^3, ,

1.
昆明医科大学第二附属医院呼吸与危重症科一病区，云南昆明　650101
2.
云南师范大学校医院，云南昆明　650101
3.
中国科学院大学宁波华美医院，浙江宁波　315000

基金项目: 国家自然科学基金资助项目（81860018）；云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目（2014FB046）

详细信息

作者简介:
刘永骏(1990～)，男，云南昆明人，医学硕士，主治医师，主要从事呼吸内科研究工作

通讯作者:
董昭兴，E-mail：dongzxkm@foxmail.com

中图分类号: R563.1⁺3
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出版历程
- 收稿日期: 2021-06-15
- 网络出版日期: 2021-09-27
- 刊出日期: 2021-09-25

The Effect of Hydrogen-Regulated Autophagy on Pulmonary Fibrosis in Paraquat Poisoned Rats

1.
Dept of Respiratory and Critical Care，The 2nd Affiliated Hospital of Kunming Medical University，Kunming Yunnan 650101
2.
Hospital of Yunnan Normal University， Kunming Yunnan 650101
3.
University of Chinese Academy of Sciences，Ningbo Huamei Hospital, Ningbo Zhejiang 315000，China

摘要

摘要: 目的探讨氢气水通过自噬对百草枯诱导的大鼠肺纤维化的治疗作用及机制。方法 6～8周SD大鼠随机分为对照组、染毒组、氢气水治疗组、雷帕霉素治疗组、综合治疗组，每组10只大鼠，在28 d处死大鼠，取肺组织。予HE染色和Masson’ s染色法观察各组标本肺组织病理变化，使用RT-PCR检测Col-ImRNA和Col-IIImRNA表达，用Westernbolt方法检测P62、LC3表达。结果与对照组相比，百草枯染毒组的大鼠肺组织出现纤维化病理改变，Col-ImRNA和Col-IIImRNA表达升高（P < 0.01），P62表达升高（P < 0.001），LC3II/I值降低（P < 0.05）；与百草枯染毒组相比，氢气水治疗组大鼠肺纤维化程度减轻，Col-ImRNA和Col-IIImRNA表达下降（P < 0.05），P62表达下降（P < 0.05），LC3II/I值升高（P < 0.05）；与百草枯染毒组相比，雷帕霉素治疗组大鼠肺纤维化程度减轻，Col-ImRNA和Col-IIImRNA表达下降（P < 0.05），P62表达下降（P < 0.01），LC3II/I值升高（P < 0.01）；与百草枯染毒组相比，综合治疗组大鼠肺纤维化程度明显减轻，Col-ImRNA和Col-IIImRNA表达显著下降（P < 0.01），P62表达下降明显（P < 0.01），LC3II/I值显著升高（P < 0.001）。结论（1）百草枯诱导大鼠肺纤维化形成并抑制了自噬；（2）雷帕霉素激活自噬并减轻了百草枯诱导的大鼠肺纤维化；（3）氢气水缓解了百草枯诱导的大鼠肺纤维化并促进了自噬；（4）氢气水与雷帕霉素联合治疗百草枯诱导的肺纤维化优于二者单用。
- 肺纤维化 /
- 百草枯 /
- 氢气水 /
- 自噬
Abstract: Objective To investigate the therapeutic effect and mechanism of hydrogen water on paraquat induced pulmonary fibrosis in rats through autophagy. Methods 6-8 week=old SD rats were randomly divided into the control group, the paraquat exposure group, the hydrogen-water treatment group, the rapamycin treatment group and the comprehensive treatment group （10 rats in each group）. All rats were sacrificed after 28 days and their lung tissues were taken and stained with hematoxylin & eosin as well as Masson for histological examination. The expressions of Col-I and COL-III mRNA were detected by RT-PCR, and the expressions of P62 and LC3 were detected by Western Blot method. Results Compared with the control group, lung tissue of rats exposed to paraquat showed pathological changes of fibrosis, increased Col-ImRNA and Col-IIImRNA expression （P < 0.01）, increased P62 expression （P < 0.001）, and decreased LC3II/I value （P < 0.05）. Compared with the paraquat exposure group, the degree of pulmonary fibrosis in hydrogen water treatment group was reduced, and the expressions of Col-ImRNA and Col-IIImRNA were decreased （P < 0.05）, the expression of P62 was decreased （P < 0.05）, and the LC3II/I value was increased （P < 0.05）. Compared with the paraquat exposure group, the degree of pulmonary fibrosis in rapamycin treatment group was reduced, the expression of Col-ImRNA and Col-IIImRNA were decreased （P < 0.05）, the expression of P62 was decreased （P < 0.01）, and the LC3II/I value was increased （P < 0.01）. Compared with the paraquat poisoning group, the degree of pulmonary fibrosis in the comprehensive treatment group was significantly reduced, the expressions of Col-ImRNA and Col-IIImRNA were significantly decreased （P < 0.01）, the expression of P62 was significantly decreased （P < 0.01）, and the LC3II/I value was significantly increased （P < 0.001）. Conclusion Paraquat induces the pulmonary fibrosis and inhibites the autophagy in rats; while Rapamycin activates the autophagy and alleviates the paraquat-induced pulmonary fibrosis in rats. Hydrogen water alleviates the paraquat-induced pulmonary fibrosis and promotes the autophagy in rats. Hydrogen water combined with rapamycin is better than hydrogen water combined with rapamycin alone in the treatment of paraquat induced pulmonary fibrosis.
- Pulmonary fibrosis /
- Paraquat /
- Hydrogen water /
- Autophagy

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图 1 肺组织染色结果（× 200）

A：HE染色结果；B：Masson’ s染色结果。

Figure 1. Lung tissue staining（× 200）

下载: 全尺寸图片幻灯片

图 2 不同处理组Col-ImRNA和Col-IIImRNA的表达

A：Col-ImRNA表达情况；B：Col-III mRNA 表达情况。与百草枯染毒组比较，^*P < 0.05，^**P < 0.01。

Figure 2. Expression of Col-I mRNA and Col-III mRNA in lung tissues in different groups

下载: 全尺寸图片幻灯片

图 3 不同处理组P62的表达和LC3-II/LC3-I比值变化

A：LC3与P62 WesternBlot结果；B：P62表达情况；C：LC3-II/LC3-I比值。与百草枯染毒组比较，^*P < 0.05，^**P < 0.01，^***P < 0.001。

Figure 3. Expression of P62 and change of LC3-II/LC3-I ratio in different treatment groups

下载: 全尺寸图片幻灯片

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