高血压大鼠左室肥厚差异蛋白质组学研究

Differential Proteomics Analysis of Left Ventricular Hypertrophy in Hypertensive Rats

目的运用蛋白质组学技术探讨腹主动脉缩窄(AAC)高血压大鼠左室肥厚差异蛋白质表达及其在左室肥厚发生机制中的作用。方法 8周龄 SD 大鼠随机分为 AAC 组及假手术组,9周后行心脏超声、血流动力学、病理学检查,并对二组心肌组织行双向电泳(2DE)找出差异点,MALDI-TOF-MS 质谱鉴定蛋白质及 West-ern-blot 验证。结果 AAC 组心率、收缩压、舒张压、左室收缩末压、左室压力变化速率、室间隔厚度与左室后壁厚度及心质量、心质量/体质量较假手术组均显著增加(P<0.05),心肌细胞排列紊乱、细胞核增大、畸形、染色加深。2DE 中以浓度相差2倍以上为差异点,鉴定的21个蛋白质中,AAC 组相对于假手术组有14个蛋白质上调,4个蛋白质下调,3个蛋白为 AAC 组新出现,上调蛋白质包括:肌球蛋白轻链、心肌α肌动蛋白1蛋白原、β肌球蛋白重链、3磷酸甘油醛脱氢酶、线粒体核糖体蛋白 L15、G 蛋白偶联受体34、酪氨酸蛋白激酶 ZAP-70、RIKENcDNA 9030617003、天冬氨酸 tRNA 合成酶等,下调蛋白为 H^-转运 ATP 合成酶、L-3脂酰辅酶 A 脱氢酶等,两组蛋白表达差异有统计学意义(P<0.05)。结论 AAC 大鼠心肌中参与糖酵解蛋白、信号转导通路蛋白、基因表达调控蛋白增加,最终心肌细胞结构蛋白增加导致心肌肥厚。

Objective To identify the different proteins expressed in the left ventricular hypertrophy（LVH） induced by abdominal aortic constriction（AAC） rats using proteomic approach and analyze the functions of these various proteins associated with the mechanism of hypertensive hypertrophy. Methods AAC and sham operation were performed on 8 week old male Sprague Dawley rats. Echocardiography, hemodynamie, cardiac morphological changes were investigated 9 weeks after operation. Differential proteins between AAC and sham operation rats were separated by 2DE and identified by MODI-TOF-MS. Changed protein were further verified by Western-blot. Results ACC rats had significant higher, HR, SBP, DBP, LVESP, dP/dt, IVSd, PWTd, HM, HM/BM in relative to sham operated rats（P〈0.05）. Histological examination revealed that myocardial cells were disarranged and cell nuclei were enlarged, malformed and densely stained. Twenty-one differential proteins in myoeardium were identified with 14 proteins up-regulation, 4 proteins down-regulation, and 3 proteins were newly found. Myosin light polypeptide, cardiac muscle α actin 1 proprotein, β-myosin heavy chain, mitochondrial ribosomal protein L15, G protein-coupled receptor 34, tyrosine-protein kinase ZAP-70, aspartyl- and tRNA synthetase were up-regulated while ATP synthase, L-3-hydroxyacyl-Coenzyme A dehydrogenase, 3-hydroxybu-tyrate dehydrogenase were down-regulated. Conclusion Various proteins involved in glycolysis, cell signaling and communication, gene expression and regulation, myocardial remodeling were increased left ventricular hypertrophy.