4周中等强度有氧运动诱导大鼠心房肌蛋白质组差异表达的研究

doi:10.12047/j.cjap.5683.2018.102

中国应用生理学杂志 ›› 2018, Vol. 34 ›› Issue (5): 450-456.doi: 10.12047/j.cjap.5683.2018.102

4周中等强度有氧运动诱导大鼠心房肌蛋白质组差异表达的研究

彭勇¹, 史绍蓉², 黄思敏³, 郭远盘⁴, 徐哲⁵, 万莉莉⁶, 雷雄⁶

1. 南京体育学院运动健康科学系, 江苏南京 210014;
2. 湖南师范大学体育学院, 长沙 410012;
3. 肇庆学院体育与健康学院, 广东肇庆 526061;
4. 湖南外国语职业学院, 长沙 410116;
5. 湖南城市学院体育系, 益阳 413039;
6. 长沙医学院基础医学系, 湖南长沙 410219

收稿日期:2018-02-11 修回日期:2018-09-25 出版日期:2018-09-28 发布日期:2019-02-21
通讯作者: 史绍蓉,Tel:13975167236;E-mail:Shisrfl@126.com E-mail:Shisrfl@126.com
基金资助:
湖南省科技厅计划项目（2007FJ3075）；江苏省教育厅高校自然科学研究面上项目（14KJB310002）；江苏高校运动人体科学品牌专业建设工程资助项目（TAPP）

Study on 4 weeks medium intensity aeraobic exercise induce the difference expression of the proteomics in rat atrial muscle

PENG Yong¹, SHI Shao-rong², HUANG Si-min³, GUO Yuan-pan⁴, XU Zhe⁵, WAN Li-li⁶, LEI Xiong⁶

1. Department of Health Sports Science, Nanjing Sport Institute, Nanjing 210014;
2. College of Physical Education, Hunan Normal University, Changsha 410012;
3. College of Physical Education and Health, Zhaoqing University, Zhaoqing 526061;
4. Hunan College of Foreign Studies, Changsha 410116;
5. Department of Physical Education, Hunan City University, Yiyang 413039;
6. Department of Basic Medicine, Changsha Medical University, Changsha 410219, China

Received:2018-02-11 Revised:2018-09-25 Online:2018-09-28 Published:2019-02-21
Supported by:
湖南省科技厅计划项目（2007FJ3075）；江苏省教育厅高校自然科学研究面上项目（14KJB310002）；江苏高校运动人体科学品牌专业建设工程资助项目（TAPP）

摘要/Abstract

摘要： 目的：探讨中等强度有氧运动对大鼠心房肌蛋白质组及其基因差异表达的影响，为运动心脏重塑和慢性心血管疾病康复研究提供研究依据。方法：20只雄性SD大鼠按照体重随机配对分为对照组、实验组（n=10），实验组大鼠每次按照速度24 m·min^-1、持续训练40 min （负荷强度相当于60%~70% VO_2max），每周训练6 d，持续训练4周中等强度有氧运动。应用双向凝胶电泳技术（2-DE）分离心房肌蛋白质点，串联飞行时间质谱仪技术鉴定电泳结果中表达量上调≥5倍以上，下调至1/5以下的13个备选目标蛋白质点。并对其中6个目标蛋白质用逆转录-聚合酶链式反应（RT-PCR）技术检测其mRNA。结果：通过软件分析，实验组与对照组比较，其中表达量下调至20%以下的点8个，上调5倍及以上点有5个，质谱鉴定分析其中的13个蛋白质点，最终鉴定出8种蛋白质和一个分子量为54 KDa的未知蛋白，包括：丙酮酸脱氢酶E1α1、线粒体乌头酸水合酶、蛋白质二硫键异构酶A3、甲基丙二酸半醛脱氢酶、线粒体二氢硫辛酸脱氢酶、异戊酰辅酶A脱氢酶、谷胱甘肽合成酶、丝裂素活化蛋白激酶3等。RT-PCR检测结果表明，与对照组相比，4周中等强度有氧运动后，大鼠心房肌中甲基丙二酸半醛脱氢酶的mRNA表达量降低（P﹤0.05），线粒体二氢硫辛酸脱氢酶、蛋白质二硫键异构酶A3、线粒体乌头酸水合酶、谷胱甘肽合成酶的mRNA表达量降低（P>0.05）；异戊烯辅酶A脱氢酶的mRNA表达量增高（P>0.05），表明mRNA表达水平与质谱鉴定结果的变化不完全一致。结论：4周的中等强度有氧运动诱导大鼠心房肌蛋白质组发生显著变化，有13个明显变化的目标蛋白，多数为能量物质代谢酶，这些目标蛋白质的变化与其mRNA表达量的变化并不完全一致，表明中等强度运动可能影响这些目标蛋白质上游基因转录的调控，也可影响下游翻译﹑修饰等的调控，导致表达的差异变化。

关键词: 蛋白质组, 有氧运动, 大鼠, 心房肌, 质谱鉴定, 双向凝胶电泳

Abstract: Objective: To investigate the effect of moderate-intensity aerobic exercise on the differential expression of rat atrial muscle Proteomics and genes, which provide research basis for the rehabilitation of chronic cardiovascular diseases and exercise -induced cardiac remodeling research. Methods: Twenty male SD rats were randomly divided into control group and experimental group (n=10) according to body weight. Rats in the experimental group were trained (6 days per week),which lasted for 4 weeks of moderate-intensity aerobic exercise at a rate of 24 m·min^-1 for 40 min (load intensity equivalent to 60%~70% VO_2max). The proteins were separated by two dimensional gel electrophoresis, and the tandem time-of- flight mass spectrometer technique was used to identify 13 candidate target protein spots. The expression levels of these 13 protein spots were up-regulated more than 5 times or down -regulated to below 1/5. The mRNA of six target proteins were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results: By software analysis, the experimental group compared with the control group, there were 8 protein points which their expression reduced more than 4/5 and 5 protein points up-regulated more than 5 times, 13 proteins were identified by mass spectrometry protein spots, the final identification results acquired 8 proteins and a unknown protein of molecular mass 54 KDa, such as:pyruvate dehydrogenase E1α1, mitochondrial aconitate hydratase, protein disulfide isomerase A3, methylmalonic acid semialdehyde dehydrogenase, mitochondrial dihydrolipoic acid dehydrogenase, isovaleryl coenzyme A dehydrogenase, glutathione synthetase, mitogen-activated protein kinase 3 and so on. Compared with the control group, the mRNA expression of methylmalonic acid semialdehyde dehydrogenase in the atrial muscle of rats was decreased after 4 weeks of moderate aerobic exercise (P<0.05), the mRNA expression levels of mitochondria Ⅱ lipoic acid dehydrogenase, protein disulfide isomerase A3, mitochondrial aconitate hydratase and glutathione synthetase were decreased (P>0.05); The mRNA expression level of isopentenyl-CoA dehydrogenase was increased (P>0.05). The results indicated that the mRNA expression level was not completely consistent with the changes in mass spectrometry identification results. Conclusion: The 4 weeks moderate-intensity aerobic exercise induced ignificant changes of rats atrial muscle protemics. The majority of the 13 identified target proteins in this experiment are energy metabolism enzymes. The majority of the expression of the target protein and the mRNA expression in the atrial muscle is inconsistent and different. Exercise may affect the regulation of gene transcription or downstream translation and modification of these target proteins, resulting in the change of differential expression.

Key words: proteomics, aerobic exercise, rat, myocardial energy metabolism, mass spectrometry 2-DE

彭勇, 史绍蓉, 黄思敏, 郭远盘, 徐哲, 万莉莉, 雷雄. 4周中等强度有氧运动诱导大鼠心房肌蛋白质组差异表达的研究[J]. 中国应用生理学杂志, 2018, 34(5): 450-456.

PENG Yong, SHI Shao-rong, HUANG Si-min, GUO Yuan-pan, XU Zhe, WAN Li-li, LEI Xiong. Study on 4 weeks medium intensity aeraobic exercise induce the difference expression of the proteomics in rat atrial muscle[J]. CJAP, 2018, 34(5): 450-456.

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4周中等强度有氧运动诱导大鼠心房肌蛋白质组差异表达的研究

Study on 4 weeks medium intensity aeraobic exercise induce the difference expression of the proteomics in rat atrial muscle

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