二氮嗪后处理对大鼠缺血再灌注心肌损伤和糖原合成酶激酶-3β表达的影响  

Effects of diazoxide-postcondition on myocardial ischemia reperfusion injury and expression of glycogen synthase kinase-3 βin rats

作　　者：赵其宏[1] 张颖[1] 栾恒飞[2] 叶英[2] 曾因明[2] 

机构地区：[1]蚌埠医学院第一附属医院麻醉科,安徽蚌埠233004 [2]徐州医学院江苏省麻醉学重点实验室

出　　处：《上海医学》2014年第12期1028-1031,I0001,共5页Shanghai Medical Journal

摘　　要：目的探讨激活磷脂酰肌醇-3-激酶(PI3K)/蛋白质丝氨酸苏氨酸激酶(Akt)信号通路对二氮嗪后处理大鼠缺血再灌注(I/R)心肌保护作用的影响及其机制。方法60只雄性Sprague-Dawley大鼠随机分入假手术组、I/R组、二氮嗪后处理组(予二氮嗪7mg/kg)、渥曼青霉素组(予渥曼青霉素15μg/kg)和二氮嗪+渥曼青霉素组(复合组)。结扎左冠状动脉前降支30min、再开放120min建立心肌I/R损伤模型。再灌注末,采用比色法检测血浆乳酸脱氢酶(LDH)、肌酸激酶(CK)表达水平,应用电子显微镜观察心肌超微结构变化,采用比色法测定心肌组织内半胱氨酸天冬氨酸特异性蛋白酶-3(Caspase-3)活性,Western免疫印迹试验检测糖原合成酶激酶-3β(GSK-3β)和磷酸化GSK-3β(p-GSK-3β)的表达水平。结果I/R组、二氮嗪后处理组、渥曼青霉素组、复合组的LDH、CK表达水平,心肌组织内Caspase-3的活性值均显著高于假手术组(P值均<0.01);二氮嗪后处理组和复合组均显著低于I/R组(P值均<0.01);复合组均显著高于二氮嗪后处理组(P值均<0.05);I/R组与渥曼青霉素组间的差异均无统计学意义(P值均>0.05)。二氮嗪后处理组和复合组的p-GSK-3β表达水平显著高于假手术组和I/R组(P值均<0.01),复合组显著低于二氮嗪后处理组(P<0.01),假手术组、I/R组和渥曼青霉素组间的差异均无统计学意义(P值均>0.05)。结论二氮嗪后处理通过激活PI3K/Akt信号通路减轻大鼠心肌I/R损伤的机制与p-GSK-3β的表达增加有关。Objective To study the role of phosphatidylinositol 3-kinase/protein serine threonine kinase （PI3K/Akt） signal pathway in myocardial preservation induced by diazoxide-postcondition against ischemia reperfusion in rats. Methods Sixty male Sprague-Dawley rats were randomly divided into five groups： sham operation group, ischemia reperfusion group, diazoxide group （7 mg/kg diazoxide）, wortmannin group （15 μg/kg wortmannin） and diazoxide＋ wortmannin group. Myocardial ischemia reperfusion injury was induced by ligating left anterior descending coronary artery for 30 min followed by reperfusion for 120 min. At the end of reperfusion, plasma lactate dehydrogenase （LDH） and creatine kinase （OK） were measured by colorimetry. Myocardial ultrastructure was observed under electron microscope. The activity of cysteinyl aspartate specific protease 3 （Caspase-3） in myocardium was detected by colorimetry. The expression of glycogen synthase kinase-3β （GSK- 313） and phosphorylation of GSK-3β （p-GSK-I3） were measured by western blotting analysis. Results The expression of LDH and OK and the activity of caspase-3 in myocardium in the sham operation group were significantly lower than those in the other four groups （all P〈0.01 ） ; the above-mentioned values in the diazoxide group and diazoxide＋ wortmannin group were significantly lower than those in the ischemia reperfusion group （all P 〈0.01 ） ; the values in the diazoxide ＋ wortmannin group were significantly higher than those in the diazoxide group（all P〈0.05）; but there were no significant differences between ischemia reperfusion group and wortmannin group （all P〉0.05）. The expression of p-GSK-3β were decreased significantly in the diazoxide group and diazoxide＋ wortmannin group as compared with sham operation group and ischemia reperfusion group （all P 〈0.01 ）; the expression of p-GSK-3β in the diazoxide ＋ wortmannin group was significantly lower than that in the diazoxide group （ P〈0.01） ; but

关 键 词：心肌再灌注损伤 二氮嗪 磷脂酰肌醇3-激酶 蛋白质丝氨酸苏氨酸激酶 糖原合成酶激酶-3Β 

分 类 号：R614[医药卫生—麻醉学]