有氧运动抑制炎症反应改善ApoE-/-动脉粥样硬化小鼠心肌纤维化机制研究

doi:10.12114/j.issn.1007-9572.2023.0516

摘要/Abstract

摘要： 背景动脉粥样硬化（AS）会引发心肌梗死、心肌纤维化等心血管病变，随着全球人口老龄化加重，发病人群逐渐增多。炎症反应是心肌纤维化的关键因素，规律的有氧运动可以减轻炎症保护心肌功能。但有氧运动对AS心肌纤维化的保护机制尚不清楚。目的本研究旨在探讨有氧运动对AS小鼠心肌纤维化的影响机制。方法 2022年2—8月选取27只8周龄的雄性ApoE-/-小鼠作为实验对象，将小鼠随机分为对照组、模型组和有氧运动组，每组9只。制备AS小鼠模型，对小鼠进行运动训练，Masson染色、苏木素-伊红（HE）染色观察心肌组织情况，蛋白质印迹法（Western blotting）检测心肌组织NOD受体3（NLRP3）、白介素1β（IL-1β）、转化生长因子β1（TGF-β1）蛋白表达情况，检测超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）表达量。结果 Masson染色结果示模型组心肌组织纤维化较对照组明显加重；有氧运动组心肌组织纤维化较模型组明显改善。HE染色结果示模型组小鼠心肌细胞排列较错乱，细胞形态、大小异常，细胞间隙增大，存在炎性细胞浸润；有氧运动组小鼠心肌细胞排列尚整齐，细胞形态、大小异常，细胞间隙基本正常。模型组NLRP3、IL-1β、TGF-β1蛋白表达高于对照组，有氧运动组NLRP3、IL-1β、TGF-β1蛋白表达低于模型组、高于对照组（P<0.05）。模型组SOD、GSH-Px表达量低于对照组，有氧运动组SOD、GSH-Px表达量高于模型组、低于对照组（P<0.05）。结论有氧运动明显改善ApoE-/- AS小鼠心肌纤维化，其机制可能与抑制心肌炎性反应、激活抗氧化水平有关。

关键词: 动脉粥样硬化, 纤维化, 有氧运动, 炎症, ApoE^-/-小鼠

Abstract:

Background

Atherosclerosis (AS) causes cardiovascular diseases such as myocardial infarction and myocardial fibrosis. Incidence of the population gradually increases with the aging of the global population. The inflammatory response is a key factor in myocardial fibrosis, and regular aerobic exercise can reduce inflammation and protect myocardial function. However, the protective mechanism of aerobic exercise against AS myocardial fibrosis is unclear.

Objective

To investigate the mechanism of the effect of aerobic exercise on myocardial fibrosis in AS mice.

Methods

From February to August 2022, twenty-seven 8-week-old male ApoE^-/- mice were selected as the experimental subjects and randomly divided into the control group, model group and aerobic exercise group, with 9 mice in each group. The mouse model of AS was prepared, and the mice were trained with exercise. The myocardial tissue was observed by hematoxylin-eosin (HE) and Masson staining. The protein expressions of NOD receptor 3 (NLRP3), interleukin1β (IL-1β) and transforming growth factor β1 (TGF-β1) in myocardial tissue were detected by Western blotting. The expressions of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected.

Results

Masson staining results showed that myocardial fibrosis in the model group was significantly worse than that in the control group. Myocardial fibrosis in the aerobic exercise group was significantly improved compared with the model group. HE staining showed that myocardial cells in the model group were disorganized, with abnormal cell morphology and size, enlarged cell gaps and inflammatory cell infiltration. Cardiomyocytes of mice in the aerobic exercise group were still neatly arranged, with abnormal cell morphology and size, and the cell gap was basically normal. The expressions of NLRP3, IL-1β and TGF-β1 in the model group were higher than those in the control group, while the expressions of NLRP3, IL-1β and TGF-β1 in the aerobic exercise group were lower than those in the model group, and higher than the control group (P<0.05). The expressions of SOD and GSH-Px in the model group were lower than those in the control group, and the expressions of SOD and GSH-Px in the aerobic exercise group were higher than those in the model group, and lower than those in the control group (P<0.05) .

Conclusion

Aerobic exercise significantly improved myocardial fibrosis in ApoE^-/- AS mice, and the mechanism may be related to inhibiting myocardial inflammatory response and activating antioxidant levels.

Key words: Atherosclerosis, Fibrosis, Aerobic exercise, Inflammation, ApoE^-/- mice

秦芳,马甜甜,于子夫,等. 有氧运动抑制炎症反应改善ApoE^-/-动脉粥样硬化小鼠心肌纤维化机制研究[J]. 中国全科医学, 2024, 27(05): 557-562. DOI: 10.12114/j.issn.1007-9572.2023.0516.
QIN Fang,MA Tiantian,YU Zifu, et al. Study on Mechanism of Myocardial Fibrosis in ApoE^-/- Atherosclerotic Mice Inhibited by Aerobic Exercise[J]. Chinese General Practice, 2024, 27(05): 557-562. DOI: 10.12114/j.issn.1007-9572.2023.0516.

图/表 5

图1 心肌组织Masson染色结果（400×）注：A为对照组，B为模型组，C为有氧运动组。

Figure 1 Results of Masson staining of myocardial tissue

图2 HE染色心肌组织形态变化（400×）注：A为对照组，B为模型组，C为有氧运动组。

Figure 2 HE staining results of morphological changes in heart tissues

表1 3组小鼠心肌组织中NLRP3、IL-1β、TGF-β1蛋白表达情况（±s）

Table 1 Expression of NLRP3，IL-1β，TGF-β1 protein in myocardial tissue of 3 groups of mice

组别	只数	NLRP3	IL-1β	TGF-β1
对照组	9	1.123±0.110	1.136±0.119	1.162±0.184
模型组	9	5.741±0.220^ab	6.447±0.524^ab	7.906±0.658^ab
有氧运动组	9	2.851±0.279^a	3.724±0.140^a	2.624±0.244^a
F值		353.02	201.52	215.98
P值		<0.001	<0.001	<0.001

图3 3组小鼠NLRP3、IL-1β、TGF-β1蛋白检测结果注：NLRP3=NOD受体3，IL-1β=白介素1β，TGF-β1=转化生长因子β1；A为NLRP3蛋白，B为IL-1β蛋白，C为TGF-β1蛋白；a为对照组，b为模型组，c为有氧运动组。

Figure 3 Results of NLRP3，IL-1β，TGF-β1 protein detection in 3 groups of mice

表2 3组小鼠心肌组织SOD、GSH-Px表达量比较（±s，U/mg）

Table 2 Comparison of SOD and GSH-Px expression in myocardial tissue of mice in 3 groups

组别	只数	SOD	GSH-Px
对照组	9	32.325±0.754	25.464±1.067
模型组	9	12.879±0.133^ab	8.672±1.056^ab
有氧运动组	9	21.615±0.432^a	15.990±0.608^a
F值		314.13	729.53
P值		<0.001	<0.001

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