Peng Yong, Shi Shao-Rong, Huang Si-Min, Guo Yuan-Pan, Xu Zhe, Wan Li-Li, Lei Xiong
Department of Health Sports Science, Nanjing Sport Institute, Nanjing 210014.
College of Physical Education, Hunan Normal University, Changsha 410012.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2018 May 8;34(5):450-456 463. doi: 10.12047/j.cjap.5683.2018.102.
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.
Twenty male SD rats were randomly divided into control group and experimental group (=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 for 40 min (load intensity equivalent to 60%~70% VO). 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).
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 (<0.05), the mRNA expression levels of mitochondria Ⅱ lipoic acid dehydrogenase, protein disulfide isomerase A3, mitochondrial aconitate hydratase and glutathione synthetase were decreased (>0.05); The mRNA expression level of isopentenyl-CoA dehydrogenase was increased (>0.05). The results indicated that the mRNA expression level was not completely consistent with the changes in mass spectrometry identification results.
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.
探讨中等强度有氧运动对大鼠心房肌蛋白质组学及基因差异表达的影响，为慢性心血管疾病康复及运动性心脏重塑研究提供理论依据。
将20只雄性SD大鼠按体重随机分为对照组和实验组（每组10只）。实验组大鼠进行训练（每周6天），持续4周的中等强度有氧运动，速度为24 m·min，持续40 min（负荷强度相当于60%~70%VO）。采用双向凝胶电泳分离蛋白质，运用串联飞行时间质谱技术鉴定出13个候选靶蛋白点，这13个蛋白点的表达水平上调超过5倍或下调至1/5以下。采用逆转录聚合酶链反应（RT-PCR）检测6个靶蛋白的mRNA。
通过软件分析，实验组与对照组相比，有8个蛋白点表达降低超过4/5，5个蛋白点上调超过5倍，质谱鉴定出13个蛋白点，最终鉴定结果获得8种蛋白质和1种分子量为54 KDa的未知蛋白质，如：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、丝裂原活化蛋白激酶3等。与对照组相比，中等强度有氧运动4周后大鼠心房肌中甲基丙二酸半醛脱氢酶的mRNA表达降低（<0.05），线粒体Ⅱ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和谷胱甘肽合成酶的mRNA表达水平降低（>0.05）；异戊烯基辅酶A脱氢酶的mRNA表达水平升高（>0.05）。结果表明mRNA表达水平与质谱鉴定结果变化不完全一致。
4周中等强度有氧运动可引起大鼠心房肌蛋白质组学显著变化。本实验鉴定出的13个靶蛋白中大多数为能量代谢酶。靶蛋白表达与心房肌中mRNA表达多数不一致且不同。运动可能影响这些靶蛋白基因转录调控或下游翻译及修饰，导致差异表达改变。
