递增负荷运动对大鼠心房肌蛋白质组研究的影响

关键词：蛋白质组  大鼠  心房肌  负荷运动  2-DE  质谱

Keywords:proteomics  rat  auricular muscle  sport  2-DE  mass-spectrometry

摘要：

      目的：通过探讨递增负荷运动对大鼠心房肌蛋白质组的影响，以筛选出心房肌组织中对运动应激具有重要意义的差异性表达的目标蛋白质，在蛋白质组学水平上研究运动对心脏功能与结构影响的规律。方法：10只雄性SD大鼠，分为运动组（n=5）和对照组（n=5）。递增负荷运动训练7周，力竭运动后6h，运动组和对照组同时麻醉处死。提取心房肌组织的全蛋白，依次对样品蛋白进行pH梯度-SDS双向凝胶电泳分离蛋白、改良考马斯亮蓝方法染色、基质辅助激光解吸离子化飞行时间串联质谱分析。结果：运动组和对照组蛋白质斑点数目分别是354±40个和382±24个；共获得具有两倍以上差异性表达的蛋白质点104个；运动后上调两倍以上的蛋白质点66个，运动后下调两倍以上的蛋白质点38个。选择差异性表达5倍以上的蛋白质点为目标蛋白进行质谱分析，共鉴定出5个蛋白质点。结论：在运动应激状态下，大鼠心房肌蛋白质发生了明显的差异性表达，这些差异性表达的蛋白质点为进一步深入运动对心脏结构与功能影响的新的目标蛋白提供了实验依据。

Abstract:

      Objective: To discuss the influence on auricular muscle proteomics after increasing progressively exercises-load-training, Select targeted protein of variability expression and important significance in sport, investigate rule of influence of sports on auricular muscle function. Method: Ten male SD rats were divided into two groups randomly： the rest group and the exercises group. After increasing progressively exercises-load-training for 7 weeks and exhausting training 6 hours, they were narcotized and executed simultaneously. Proteins of auricular muscle were extracted and separated by 2-DE, stained by Coomassie brilliant blue dye and analyzed by MALDI-TOF-TOF-MS in turn. Result: There were 354±40 and 382±24 protein spots found in the sport group and test group, matching rate were 68.67％±9.87% and 72.67％±8.62%,coefficient correlation were 0.71; In 104 differentially expressed spots after traning, there were 66 in up-regulation 2 times,including 18 in up-regulation 5 times, none protein spot increasing; 38 in down-regulation 2 times,including 23 in down-regulation 5 times. Protein spots of differential expressed more than 5 times were selected as object proteinum in mass chromatographic analysis, and 5 protein spots of which were assess. Conclusion: Experimental results indicated that auricular muscle protein were obviously variously expressed， and it was a sufficient experiment base in order to profoundly study original objective proteinum of influence of sports on auricular muscle of structure and function.