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目的:探讨力竭运动对大鼠心室肌蛋白质组表达的影响,初步筛选出对运动应激具有重要意义的心室肌目标蛋白质。方法:将10只雄性SD大鼠随机分为安静对照组和运动组(n=5)。运动组大鼠采用三级递增运动负荷跑台训练建立一次性力竭运动实验动物模型。取心室肌,提取心室肌组织全蛋白进行双向凝胶电泳(2-DE)分离。通过图像分析软件PD Quest进行分析后,选择运动后消失和表达量上调5倍或下调4/5以上的蛋白质点作为备选质谱鉴定蛋白点。结果:在电泳图谱上对照组平均检测到蛋白质点332±17个,运动组平均检测到347±22个。运动后共有47个蛋白质点发生了变化:5个点在运动后消失;表达量上调3倍以上的点有28个,下调2/3以上的点有14个,其中上调5倍以上的点14个,下调4/5以上的点4个。对6个备选蛋白质点进行了质谱鉴定,共鉴定出心肌α-肌球蛋白重链、原肌球蛋白-1、甘露糖结合蛋白C前体、腈水解酶和一个未知的分子量为21kD的蛋白质。结果表明,力竭运动后大鼠心室肌蛋白质组发生了明显变化;本研究为寻找新的运动性疲劳的生物标志物提供了新的思路。
OBJECTIVE: To investigate the effect of exhaustive exercise on the expression of ventricular muscle proteome in rats and to screen out the target protein of ventricular muscle, which is important for exercise stress. Methods: Ten male Sprague-Dawley rats were randomly divided into the control group and the exercise group (n = 5). Rats in exercise group were subjected to one-time exhaustive exercise animal model by three-stage progressive exercise load treadmill training. Take ventricular muscle, ventricular muscle extract from whole protein for two-dimensional gel electrophoresis (2-DE) separation. After analysis by PD Quest, the image analysis software PDQuest was used to select protein spots that disappeared after the exercise and whose expression levels were up-regulated by 5-fold or down by 4/5 or more as alternative mass spectrometry spots. RESULTS: On the electrophoretogram, an average of 332 ± 17 protein spots were detected in the control group and 347 ± 22 in the exercise group. A total of 47 protein spots changed after exercise: 5 spots disappeared after exercise; 28 spots were up 3 times more, 14 spots more than 2/3 down, and 5 points more than 14 spots A, 4 points down more than 4 points. Six candidate protein spots were identified by mass spectrometry, and cardiac muscle α-myosin heavy chain, tropomyosin-1, mannose binding protein C precursor, nitrilase and an unknown protein with a molecular weight of 21 kD protein. The results show that the ventricular muscle proteome changes significantly after exhaustive exercise. This study provides a new idea for finding new biomarkers of exercise-induced fatigue.