基于体外循环冠脉搭桥术中冷灌缺血心脏筛选人心肌早期缺血损伤特异生物标记的研究

项目名称： 基于体外循环冠脉搭桥术中冷灌缺血心脏筛选人心肌早期缺血损伤特异生物标记的研究

项目编号： No.30871061

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 生物科学

项目作者： 杨天德

作者单位： 中国人民解放军第三军医大学

项目金额： 31万元

中文摘要： 目前肌钙蛋白（cTn）I、T等特异性指标只能反映心肌缺血坏死的不可逆损伤，而反映心肌缺血可逆性损伤的缺血修饰白蛋白（IMA）等指标均非心肌来源，因而特异性差。本项目拟选用体外循环冠脉搭桥术中的晶体停搏液灌注缺血心脏, 从冠状静脉窦收集组份简单且不含血液成分的回流液。通过收集在心肌细胞释放其坏死标记cTnT和cTnI之前各时段经冠脉回流液，采用双向电泳、鸟枪法等技术,对回流液中的蛋白进行分离和鉴定，经数据库查询，从中筛选出与心肌细胞结构组成和功能代谢密切相关的蛋白，然后采用ELISA法分别检测这些蛋白在不同时间段留取的回流液中的浓度，筛选出释放量与心肌缺血时间呈正相关的生物标记；通过对比研究不稳定性心绞痛患者与正常人的运动实验，观察人急性心肌缺血过程中该生物标记的血液浓度变化及其与IMA和cTnI变化的关系，从而筛选出特异性高且敏感性好的心肌缺血生物标记，为最终实现临床早期特异诊断心肌缺血损伤打下基础。

中文关键词： 生物标记;人心肌缺血;冷灌缺血心脏;体外循环;

英文摘要： It is well known that cardiac troponin (cIn) I and cTnT are specific indexes of myocardial ischemic necrosis detecting irreversible cardiac injury . But the known indexes detecting reversible cardiac injury such as ischemia modified albumin (IMA) are poor specificity because of not origin of myocardium. The study will collect the return flow with simple components from coronary sinus during coronary artery surgery under cardiopulmonary bypass with cold crystalloid cardioplegia solution, which will be separated and identified by the technologies of proteoms such as two-dimensional electrophoresis (2-DE) , shot-gun and mass spectrometry. Firstly,the proteins closely related to structure composition, function and mebabolism of myocardium will be found out from those identified proteins by databass query. Secondly , proteins with the release amount positively related to the time of myocardial ischemia will be picked out by ELISA. Thirdly, the blood concentration of those proteins will be detected in both the health and the patients with unstable angina pectoris and compared with that of IMA and cTnI. Finally, biological markers with good specificity and high sensitivity will be obtained, which lay a good foundation for eraly and specific diagnosis of cardiac ischemia in clinic.

英文关键词： biological markers;human cardiac ischemia;cold crystalloid cardioplegia solution;cardiopulmonary bypass