摘要
目的探讨腹主动脉狭窄诱导大鼠心肌肥大后心肌组织蛋白质表达谱的变化。方法健康雄性Wistar大鼠12只,随机分为模型组和对照组(均n=6)。模型组行腹主动脉缩窄术,对照组仅分离腹主动脉不行缩窄术。术后4周结束实验并提取左心室组织蛋白质,双向电泳分离心肌组织蛋白质,分析差异显示的蛋白质并选取9个差异显著的蛋白点进行胶内酶切和质谱分析。结果双向电泳可分离(454±13)个蛋白质,点匹配率为(78.7±1.4)%,心肌组织13种蛋白质出现明显差异表达,与对照组比较,模型组4种蛋白质表达上调,9种蛋白质表达下调。质谱分析鉴定出7个蛋白质为:与能量代谢相关的丙酮酸脱氢酶E1和β-烯醇酶、与细胞收缩功能相关的α-肌球蛋白重链和心脏α-肌动蛋白前体、与甲状腺激素代谢相关的甲状腺素结合蛋白等5种蛋白质在模型组表达下调,肌球蛋白轻链和与细胞内源性保护相关的热休克蛋白B6等2种蛋白质在模型组表达上调。结论腹主动脉缩窄致大鼠心肌肥大后,心肌组织蛋白质组变化显著,这些变化涉及与心肌细胞能量代谢、细胞骨架以及心肌收缩等有关的几组蛋白质,提示上述蛋白质组改变可能参与了心肌肥大过程。
Objective To explore proteomic changes in rat hypertrophic myocardium induced by abdominal aortic constriction. Methods Healthy male Wistar rats were randomly divided into model group and control group（ n = 6）, and the rats in model group were operated on abdominal aortic constriction. After extracting protein of left ventricular tissue at 4 weeks after surgery, 2-D electrophoresis（2-DE） was employed to separate proteins, and 7 protein spots expressed differentially were picked out and subjected to tryptic in-gel digest and matrix-assisted laser desorption/ionization-time of flight mass spectrometry （MALDI-TOF） for identification. Results 454 ± 13 proteins were detected and the match rate was（78.7 ± 1.4） %. 13 proteins displayed significant changes in rat hypertrophic myocardium, of which 4 proteins increased and 9 proteins decreased inexpression. 7 proteins identified after mass spectrometry are as follows： pyruvate dehydrogenase El（PDH） and β- enolase associated with energy metabolism,α-actin proprotein＇and a-myosin heavy chain（α-MHC） associated with cell contraction, transthyretin（TTR） linked with thyroid hormone, the expression in all of above decreased in model group, and the expression of both myosin light chain （MLC） related with cell contraction and heat shock protein B6（HspB6） related with cell protection increased in hypertrophic myocardium. Conclusion The proteome in hypertrophyic myocardium induced by abdominal aortic constriction alters dramatically, the changes of energy metabolism, cell skeleton and contraction function in myocytes may be involved in hypertrophic process.
出处
《中国微循环》
2009年第2期84-89,共6页
Journal of Chinese Microcirculation
基金
国家自然科学基金(No.30770902)
国家自然科学基金重大国际合作(No.30620130111)
关键词
心肌肥大
蛋白质组
双向电泳
质谱
Hypertrophy
Proteome
Two-dimensional electrophoresis
Mass spectrometry
