大鼠肢体缺血预处理延迟相心肌蛋白质组学的变化  

Changes in myocardial proteomics in late phase of limb ischemic preconditioning in rats

作　　者：郑晓春[1] 高飞[1] 陈江湖[1] 涂文劭[1] 张家平[1] 樊梦莹 曾星星[1] 

机构地区：[1]福建医科大学省立临床医学院福建省立医院麻醉科,福州市350001

出　　处：《中华麻醉学杂志》2016年第4期421-426,共6页Chinese Journal of Anesthesiology

基　　金：福建省卫生系统中青年骨干人才培养项目（2014-ZQN-JC-01）;福建省卫生厅中医药科研课题（wzsy201302）;福建省自然科学基金（2013J01116）

摘　　要：目的探讨大鼠肢体缺血预处理延迟相心肌蛋白质组学的变化。方法清洁级雄性成年SD大鼠12只，8～9周龄，体重260～280g，采用随机数字表法分为2组（n=6）：肢体缺血预处理组（LIP组）和对照组（c组）。LIP组用橡皮筋环扎右后肢根部，阻断血流5rain，开放再灌注5min，共循环3次，实施肢体缺血预处理。预处理后24h时，取左心室组织，采用同位素相对标记与绝对定量技术（iTRAQ）和液相色谱一质谱法检测蛋白表达谱的差异（组间表达差异大于1．2倍且P〈0．05）。对鉴定得到的差异表达蛋白进行生物信息学分析，并用Western blot法验证部分差异表达蛋白。结果筛选得到55个差异表达蛋白，其中35个蛋白表达上调，20个蛋白表达下调。生物信息学分析发现，这些蛋白多为细胞器、细胞膜及大分子复合物，广泛参与代谢、生物调节、应激反应及信号传导等过程，具有分子结合活性、催化活性、抗氧化及调节酶活性等功能。Western blot法的验证结果与iTRAQ分析结果一致。结论肢体缺血预处理延迟相可诱发与心肌细胞调节能量代谢、抗氧化、调控基因表达及参与蛋白质折叠与降解等有关的55个蛋白表达变化，这可能是其心肌保护效应的机制。Objective To investigate the changes in myocardial proteomics in the late phase of limb ischemic preconditioning （LIP） in rats. Methods Twelve pathogen-free adult male Sprague-Dawley rats, aged 8-9 weeks, weighing 260-280 g, were randomly assigned into LIP group （n= 6） and control group （group C, n= 6） using a random number table. Limb ischemia was preceded by 3 cycles of 5-rain ischemia which was induced by ligation of the root of the right hindlimb with a rubber band followed by 5-min reperfusion in group LIP. At 24 h after LIP, the tissues were obtained from the left ventricle, and the isobaric tags for relative and absolute quantification technique and liquid chromatography-mass spectrometry were applied to detect the differences in protein expression profiles between the two groups （the difference in expression between the two groups〉1.2 times and P〈0.05）. The identified differentially expressed proteins were analyzed using the bioinformatics, and some were further verified by Western blot. Results A total of 55 proteins were identified to be differentially expressed, and among the 55 proteins, the expression of 35 proteins was up-regulated, and the expression of 20 proteins was down-regulated. Bioinformatics analysis showed that most of the 55 proteins were organellcs, cell membrane or macromolecular compounds, were involved in the process such as metabolism, biological regulation, stress response and signal transduction, and showed functions such as the binding affinity to molecules, catalytic activity, anti-oxidant aetivity, and modulation of the activity of enzyme. The results verified by Western blot were consistent with those shown by using the isobaric tags for relative and absolute quantification analysis. Conclusion The late phase of LIP can induce changes in the expression of the 55 proteins involving regulation of energy metabolism, anti-oxidant action, regulation of gene expression, and protein folding and degradation in the rnyocardiurn, which may be the mechanism of myocardi

关 键 词：四肢 缺血预处理 心肌 蛋白质组学 

分 类 号：R614[医药卫生—麻醉学]