递增负荷运动后大鼠心房肌蛋白质组的差异性表达

目的：以往的许多研究都是针对心房的单个蛋白质进行的，没有系统地对心房的“全部”蛋白质进行研究，因而不能全面地对心房在运动性心脏重塑或运动性心脏疲劳中的作用做出解释。本实验应用双向凝胶电泳技术，认识递增运动负荷训练后，大鼠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，从蛋白质组学水平上探讨运动性疲劳的发生及发展的规律。 方法：实验于2006-11／2007-03在湖南师范大学生命科学学院蛋白质化学与蛋白质组学国家教育部重点实验室和省级运动人体科学实验室完成。①实验动物：10只雄性SD大鼠，随机分为运动组5只和对照组5只。实验过程中对动物处置符合动物伦理学要求。②实验方法：递增运动负荷训练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梯度一SDS双向凝胶电泳分离蛋白、改良考马氏亮蓝方法染色。运用Bio PD quest图像分析软件对双向凝胶电泳图谱进行分析。结果：①运动组和对照组蛋白质斑点数目分别是（354±40）个和（382±24）个，匹配率分别为（68．67±9．87）％和（72．67±8．62）％，相关系数为0．71。②共获得具有2倍以上差异性表达的蛋白质点104个。③运动后上调2倍以上的蛋白质点66个，其中运动后上调5倍以上的蛋白质点18个，未见运动后新增的蛋白质点。④运动后下调2倍以上的蛋白质点38个，其中运动后下调5倍以上的蛋白质点23个，运动后消失的蛋白质点7个。 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观察具有运动性疲劳特征的新的目标蛋白提供了充分的实验依据。

Differential expression of rat auricular muscle proteome after increasing intensity training

AIM： Previous studies focus on single protein in ventricles, but not the whole proteins, thus they could not completely explain the role of ventricles in exercise-induced heart remodeling or exercise-induced heart fatigue. Using two-dimensional gel electrophoresis, this study discussed the targeted protein of differentially expressed and important significance in auricular muscle after increasing intensity training, so as to investigate the rule of development of sports fatigue at proteome level.
METHODS： The experiment was carried out in the Key Laboratory of Protein Chemistry and Proteome, Ministry of Education, College of Human Movement Science of Hunan Normal University and Provincial Laboratory of Human Movement Science from November 2006 to March 2007. ①Ten male SD rats were randomly divided into exercise group and control group （n =5）. All the procedure was accorded with Animal Ethical Standards. ②After increasing intensity training for 7 weeks and exhausted training for 6 hours, the animals were executed under anesthesia. The whole protein of auricular muscle was extracted, separated by pH gradient-SDS two dimensional gel electrophoresis, and stained by Coomassie brilliant blue method. The electrophoregram was analyzed using Bio PD quest image analysis software.
RESULTS： ①The protein spots in the exercise group and control group were （354±40） and （382±24）, respectively; the matching rates were （68.67 ± 9.87）% and （72.67 ± 8.62）%, and the coefficient correlation were 0.71. ②104 differentially expressed spots were found. ③There were 66 protein spots in up-regulation 2 times above, including 18 in up-regulation 5 times. No protein spot was found after exercisein. ④There were 38 protein spots in down-regulation 2 times above, including 23 in down-regulation 5 times. Seven protein sports disappeared after exercise.
CONCLUSION： Auricular muscle proteins are obviously differentially expressed under sports stress. The protein spots provide sufficient experimental