草酰乙酸对心肌缺血再灌注损伤的保护作用及机制研究  

Protective Effect and Regulation Mechanism of Oxaloacetate on Myocardial Ischemia Reperfusion Injury in Rats

作　　者：丁士骜 梅举[1] 鲍春荣[1] 张俊文[1] 杨阳[1] 袁源[1] 尹航[1] 

机构地区：[1]上海交通大学医学院附属新华医院心胸外科,上海200092

出　　处：《中国胸心血管外科临床杂志》2016年第3期274-279,共6页Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

基　　金：上海市科学技术委员会资助项目(13XD1403200)~~

摘　　要：目的 探索草酰乙酸对大鼠心肌缺血-再灌注损伤的保护作用。方法 将60只体重200~250 g雄性大鼠随机分为6组：阴性对照组、假手术组、模型组、草酰乙酸预处理后心肌缺血-再灌注（OAA）模型组（三个亚组：高,中,低剂量组）。建立大鼠心肌缺血再灌注模型,造模期间记录左心室内压（LVSP）,左心室内压最大变化速率（±dp/dt_（max））以及左心室舒张期末压力（LVEDP）,左冠状动脉前降支结扎30 min后实现再灌注180 min后采血,测定肌钙蛋白Ⅰ（c Tn-Ⅰ）,乳酸脱氢酶（LDH）,超氧化物歧化酶（SOD）,谷胱甘肽过氧化物酶（GSH-Px）。取心脏组织,染色,计算梗死范围。使用WB方法检测心肌标本的NF-E2相关因子2（Nrf2）、Kelch样环氧氯丙烷相关蛋白-1（Keap1）及Nrf2下游的血红素氧合酶1（HO-1）的表达。结果 与假手术组比较,阴性对照组各项指标差异均无统计学意义（P〉0.05）;而模型组血清SOD、GSH-Px活力显著降低（P〈0.01）,血清LDH、c Tn-I含量显著升高（P〈0.01）,心肌组织梗死面积显著升高（P〈0.01）;与模型组相比,草酰乙酸预处理组血清SOD、GSH-Px活力显著升高（P〈0.05）,LDH、c Tn I含量显著降低（P〈0.05）,心肌组织梗死面积显著降低（P〈0.05）,其中高剂量组上述各项指标改变更加明显,差异有统计学意义（P〈0.01）;与模型组相比,OAA预处理组的Keap1显著下调（P〈0.001）,总Nrf2表达增加,并且Nrf2的核转移及下游HO-1表达显著上调（P〈0.001）,提示草酰乙酸可通过（至少部分可通过）Keap1-Nrf2通路减少缺血再灌注氧化应激损伤,减轻心肌损害,起到心肌保护作用。结论 草酰乙酸可通过Keap1-Nrf2通路减少缺血再灌注氧化应激损伤,对心肌缺血-再灌注损伤具有保护作用。Objective To investigate the protective effect and the regulation mechanism of oxaloacetate （OAA） on myocardial ischemia reperfusion injury in rats. Methods Sixty rats, weight ranged from 200 to 250 grams, were randomly divided into 6 groups： a negative control group, a sham operation control group, a model control group, an OAA pretreatment myocardial ischemia-reperfusion model group （three subgroups： 15 mg/kg, 60 mg/kg, 240 mg/kg）. We established the model of myocardial ischemia reperfusion of rats and recorded the internal pressure of left ventricle （LVSP）, the maximal rate of left ventricular pressure change （＋dp/dtmax） and left ventricular end diastolic pressure （LVEDP）. We restored reperfusion 180 minutes after ligating the left anterior descending coronary artery 30 minutes and determinated cardiac troponin I （cTn-I）, lactate dehydrogenase （LDH）, superoxide dismutase （SOD）, glutathione peroxidase （GSH-Px）. We took out heart tissues, stained it and calculated the infarcted size. We used the Western blot to detect the expression of NF-E2 related factor 2 （Nrf2）, Kelch-like ECH-associated protein-1 （Keapl） and heme oxygenase-1 （HO-1） . Results Compared with the sham operation group, heart function indexes in the negative control group had no significant difference （P〉0.05）. But in the model control group there was a decrease （P〈0.05） And the serum levels of LDH, cTn-I, and myocardial infarcted size were significantly increased （P〈0.01）. Compared with the model control group, heart function indexes in the OAA pretreatment groups improved, the serum LDH, cTn-I activity, and infarct size decreased （P〈0.05）, SOD and GSH-Px activity increased （P〈0.05）. And these results were statistically different （P〈0.01） in the high dose OAA pretreatment groups. Compared with the model control group, the expression of Keapl in the OAA pretreatment group was down-regulated （P〈0.001） while total Nrf2, nucleus Nrf2 and its downstream HO

关 键 词：草酰乙酸(OAA) 心肌缺血再灌注损伤 Kelch样环氧氯丙烷相关蛋白-1(Keap1) NF-E2相关因子2(Nrf2) 血红素氧合酶1(HO-1) 

分 类 号：R541[医药卫生—心血管疾病]