心肌缺血早期表达蛋白的质谱研究

目的用SELDI-TOFMS观察大鼠急性心肌缺血后心肌蛋白质谱的变化规律,筛选心肌缺血的蛋白标记物,为法医学心源性猝死的诊断提供依据。方法SD大鼠分成手术组,假手术组和非手术对照组。在结扎大鼠冠脉前降支后5min～6h内不同的时间段取大鼠心肌组织。蛋白提取后用CM10芯片结合SELDI-TOFMS质谱仪检测。结果血清CM10芯片检测到3个缺血后蛋白峰,质荷比(m/z)分别为6304Da、8337Da、8375Da,在手术组中阳性率和特异性均为100%,假手术组和非手术组中均没有发现。特异性升高的峰有3个:峰6658Da、6876Da,都是在实验组中≥15min以后升高2倍以上,8577Da峰则在缺血≥5min组升高。结论大鼠心肌组织中6304Da、8337Da、8375Da蛋白峰都是急性心肌缺血后的较为特异性改变,可在心肌缺血后5min内出现,有可能做为心肌缺血的蛋白标记,亦可能用于心肌缺血和法医学心源性猝死的诊断。

Proteomic study of early expressed protein of myocardial ischemia

Objective To observe the protein change and find potential biomarkers of myocardial ischemia in a rat model using surface-enhanced laser desorption /ionization time of flight mass spectrometry(SELDI-TOF MS).Such research may be useful for the diagnosis of acute cardiac ischemia and sudden cardiac death.Methods Sprague-Dawley rats were divided as three groups:operation group,sham operation group,non-operation group.The myocardiums were collected 5min~6h after the coronary artery ligation.Protein extracts were processed by CM10 ProteinChip and analyzed by SELDI-TOF MS.Results Three novel ischemia related peaks with m /z 6304Da,8337Da and 8375Da were only found in operation groups with the 100% positive rate and 100% specificity.Three ischemia related peak height increasing peaks were found:two of them,peak 6658Da and 6876Da increased in ≥15min groups,while peak 8577Da increased in ≥5 min groups.Conclusion Three novel myocardial ischemia related peaks 6304Da、8337Da、8375Da are specific for the myocardium ischemia.They appeared as early as 5 min after the onset of LAD liagation and may serve as the bismarkers of acute myocardial ischemia or ancillary ischemia sign for SCD.