JNK和p38MAPK信号通路在吗啡后处理减轻大鼠心肌再灌注损伤中的作用：离体实验  

Role of JNK and p38MAPK signaling pathways in attenuation of myocardial ischemia-reperfusion injury by morphine postconditioning： an in vitro experiment

作　　者：申恒花[1] 陈作雷[2] 唐润栋 刘英志[2] 褚海辰[2] 

机构地区：[1]青岛市第八人民医院麻醉科,266003 [2]青岛大学附属医院麻醉科,266071

出　　处：《中华麻醉学杂志》2016年第11期1349-1352,共4页Chinese Journal of Anesthesiology

摘　　要：目的评价c-Jun氨基末端激酶（JNK）和p38丝裂原活化蛋白激酶（p38MAPK）信号通路在吗啡后处理减轻心肌再灌注损伤中的作用。方法健康成年雄性SD大鼠，体重180～240 g，采用Langendorff灌注装置建立大鼠离体心脏灌注模型。平衡灌注15 min后，取离体心脏52个，采用随机数字表法分为4组（n＝13）：对照组（C组）K-R液持续灌注105 min；缺血再灌注组（I/R组）停止K-R液灌注造成全心缺血45 min，恢复K-R液灌注60 min；吗啡后处理组（MP组）缺血45 min，再灌注即刻灌注含3.0 μmol/L吗啡的K-R液10 min，然后灌注K-R液50 min；吗啡后处理＋茴香霉素组（MP＋A组）缺血45 min，再灌注即刻灌注含3.0 μmol/L吗啡和1.0 μmol/L茴香霉素（JNK和p38MAPK的激动剂）的K-R液10 min，然后灌注K-R液50 min。再灌注60 min时每组取8个心脏，测定心肌CK-MB释放量，观察心肌梗死情况，计算心肌梗死体积。再灌注20 min每组取5个心脏，采用Western blot法测定心肌组织磷酸化JNK（p-JNK）、磷酸化p38MAPK（p-p38MAPK）和细胞色素c （Cyt c）的表达水平，分光光度法检测烟酰胺腺嘌呤二核苷酸（NAD＋）含量。结果与C组比较，I/R组、MP组和MP＋A组心肌梗死体积和心肌CK-MB释放量增加，心肌组织p-JNK、p-p38MAPK和Cyt c的表达上调，NAD＋含量降低（P〈0.05）；与I/R组比较，MP组和MP＋A组心肌梗死体积和心肌CK-MB释放量减少，MP组心肌组织p-JNK、p-p38MAPK和Cyt c表达下调，NAD＋含量升高（P〈0.05）；与MP组比较，MP＋A组心肌梗死体积和心肌CK-MB释放量增加，心肌组织p-JNK、p-p38MAPK和Cyt c的表达上调，NAD＋含量降低（P〈0.05）。结论吗啡后处理减轻大鼠心肌缺血再灌注损伤的机制与抑制JNK和p38MAPK信号通路的激活有关。ObjectiveTo evaluate the role of c-Jun N-terminal kinase （JNK） and p38 mitogen-activated protein kinase （p38MAPK） signaling pathways in attenuation of myocardial ischemia-reperfusion （I/R） injury by morphine postconditioning.MethodsHealthy adult male Sprague-Dawley rats, weighing 180-240 g, were used in the study.Their hearts were excised and retrogradely perfused in a Langendorff apparatus with Krebs-Ringer （K-R） buffer saturated with 95% O2 -5% O2 at 37 ℃.After 15 min of equilibration, 52 isolated hearts were divided into 4 groups （n=13 each） using a random number table： control group （group C）, I/R group, morphine postconditioning group （group MP）, and morphine postconditioning plus anisomycin group （group MP＋ A）. The hearts were continuously perfused with K-R buffer for 105 min in group C. In group I/R, the hearts were subjected to 45 min of global ischemia by stopping perfusion with K-R buffer, followed by 60 min of reperfusion by restoration of perfusion with K-R buffer.In group MP, the hearts were subjected to 45 min of global ischemia, followed by 10 min of reperfusion with K-R buffer containing 3.0 μmol/L morphine and then by 50 min of reperfusion with K-R buffer.In group MP＋ A, the hearts were subjected to 45 min of global ischemia, followed by 10 min of reperfusion with K-R buffer containing 3.0 μmol/L morphine and 1.0 μmol/L anisomycin （an activator of JNK and p38MAPK） and then by 50 min of reperfusion with K-R buffer.At 60 min of reperfusion, 8 hearts in each group were selected for measurement of the myocardial infarction and amount of creatine kinase-MB （CK-MB） released from the myocardium, and the myocardial infarct size was calculated.At 20 min of reperfusion, 5 hearts in each group were selected to detect the expression of phosphorylated JNK （p-JNK）, phosphorylated p38MAPK （p-p38MAPK） and cytochrome c （Cyt c） in myocardial tissues （by Western blot） and content of nicotinamide adenine dinucleotide （NAD＋ ） in myocardial tissu

关 键 词：JNK丝裂原活化蛋白激酶类 P38丝裂原活化蛋白激酶类 信号传导 吗啡 缺血后处理 心肌再灌注损伤 

分 类 号：R614[医药卫生—麻醉学]