心肌缺血小型猪模型差异蛋白质组学研究

Quantitative and comparative proteomics analysis of pig models with chronic myocardial Ischemia

目的:筛选心肌缺血相关的蛋白质,以期发现心肌缺血特征性的差异蛋白质或蛋白质群。方法:对模型动物小型猪施行冠脉Ameroid环缩术,通过对模型动物的动态观察,综合评价,并参照WHO冠心病心绞痛诊断标准,明确术后28天符合心肌缺血诊断。术后28天,动物前腔静脉取血,血样处理后进行荧光差异蛋白电泳(2D-DIGE),对差异蛋白点进行MALDI-TOF/TOF分析,获取蛋白样品的肽质量指纹图。结果:共筛选出31个差异表达蛋白质点,其中17个蛋白质点在4周模型组中下调,14个蛋白点上调。对其中15个蛋白点采用质谱技术成功鉴定。白蛋白、血红素蛋白、烟酸受体在4周模型组中低表达,CH4 and secrete domain of Swine IgM、mutated immunoglobulin heavy chain、肌球蛋白、磷脂酶C、白细胞抗原相关酪蛋白磷酸酶相关蛋白、磷酸核糖焦磷酸相关蛋白-1、Ig gamma 4 chain constant region在4周模型组中高表达。结论:初步发现碱性磷酸酶、肌球蛋白、白细胞抗原相关酪蛋白磷酸酶相关蛋白、磷酸核糖焦磷酸合成酶相关蛋白1、血红素蛋白、烟酸受体可能与心肌缺血发生发展相关。

Objective：To explore the differences in protein with animal model of myocardial ischemia. Methods：Healthy pigs were used in the experiment. After thoracotomy and pericardieetomy, ameroid constrictor was implanted around the left anterior descending artery. After operation, observation was taken continuously. Then the chronic myocardial isehemia was evaluated by coronary arteriog raphy and electrocardiogram. It is proved that 28 days after operation is the point to stable stage of myocardial ischemia with the reference to WHO diagnostic criteria. The plasma samples of model group and control group were extracted from the the superior vena cava and examined by the fluorescence differential in-gel eleetrophoresis （2D-DIGE） technique. Protein spots detected differential with statistical significance were identified by MALDI-TOF/TOF. Results：Intensity changes of 31 spots were detected with statistical significance. 17 protein spots of them down-regulated otherwise 14 up-regulated in model group. And 15 of them were identified by MALDI-TOF/TOF successfully. Several proteins down-regulated such as albumin, hemopexin, nicotinic acid receptor. While, CH4 and secrete domain of Swine IgM,mutated immunoglobulin heavy chain, myosin heavy chain,Ppfibpl ,PRPS-assoeiated protein 1 ,Ig gamma 4 chain constant region were up-regulated. Conclusion：It is proved that alkaline phosphatase, myosin heavy chain,Ppfibpl ,PRPS-assoeiated protein 1, hemopexin, nicotinic acid receptor are possibly the specific proteins for chronic myocardial ischemia. But the conclusion needs further validation and mechanism studies.