ATP敏感性钾通道在异氟醚预处理减轻大鼠脑缺血再灌注损伤中的作用：与JNK信号通路的关系  

Role of mitochondrial ATP-sensitive potassium channel in mitigation of cerebral ischemia-reperfusion injury by isoflurane preconditioning in rats： the relationship with JNK signaling pathway

作　　者：章放香[1] 张竟超[1] 罗云鹏[1] 赵倩[1] 张伟晶[1] 刘承铭 邱冰 

机构地区：[1]贵州省人民医院麻醉科,贵阳市550001 [2]贵州省人民医院急诊科,贵阳市550001 [3]贵州省骨科医院骨科

出　　处：《中华麻醉学杂志》2014年第11期1389-1392,共4页Chinese Journal of Anesthesiology

基　　金：贵州省科技厅社会发展科技攻关项目[黔科合SY（2012）3045]

摘　　要：目的 评价线粒体ATP敏感性钾通道在异氟醚预处理减轻大鼠脑缺血再灌注损伤中的作用及其与c-Jun氨基末端激酶（JNK）信号通路的关系.方法 清洁级健康成年雄性SD大鼠32只,体重280 ～ 320 g,采用随机数字表法分为4组（n=8）：假手术组（S组）、脑缺血再灌注组（I/R组）、异氟醚预处理组（Ⅰ-pre组）和5-羟葵酸组（5-HD组）.I/R组采用改良Pulsinelli四血管阻断法制备大鼠全脑缺血再灌注模型;Ⅰ-pre组在缺血前每天吸入1.5％异氟醚1h,连续5 d;5-HD组处理同Ⅰ-pre组,在缺血处理前30 min腹腔注射5-羟癸酸15 mg/kg.于再灌注24 h时行神经行为学评分,于再灌注72h时处死大鼠取海马组织,采用TUNEL法检测凋亡神经元,计算神经元凋亡率,免疫组化法检测caspase-3表达,Westem blot法检测磷酸化JNK （p-JNK）蛋白表达.结果 与S组比较,I/R组、Ⅰ-pre组和5-HD组格子爬行数减少,悬挂时间缩短,海马神经元凋亡率升高,caspase-3表达上调,I/R组和5-HD组p-JNK蛋白表达上调（P＜0.05）,Ⅰ-pre组p-JNK蛋白表达差异无统计学意义（P＞0.05）;与I/R组比较,Ⅰ-pre组格子爬行数增多,悬挂时间延长,海马神经元凋亡率降低,caspase-3和p-JNK蛋白表达下调（P＜0.05）,5-HD组上述指标差异无统计学意义（P＞0,05）;与Ⅰ-pre组比较,5-HD组格子爬行数减少,悬挂时间缩短,海马神经元凋亡率升高,caspase-3和p-JNK蛋白表达上调（P＜0.05）.结论 线粒体ATP敏感性钾通道可通过阻断JNK信号通路参与异氟醚预处理减轻大鼠脑缺血再灌注损伤的过程.Objective To evaluate the role of mitochondrial ATP-sensitive potassium （mitoKATP） channel in mitigation of cerebral ischemia-reperfusion （I/R） injury by isoflurane preconditioning in rats and the relationship with c-Jun N-terminal kinase （JNK） signaling pathway.Methods Thirty-two male Sprague-Dawley rats,weighing 280-320 g,were randomly divided into 4 groups （n =8 each） using a random number table：sham operation group （group S）,group I/R,isoflurane preconditioning group （group Ⅰ-pre）,and 5-hydroxydecanoate （5-HD,a selective mitoKATP channel antagonist） group.Cerebral I/R was produced by modified 4-vessel technique described by Pulsinelli in anesthetized rats.In group Ⅰ-pre,the rats were exposed to 1.5％ isoflurane for 1 h everyday for 5 consecutive days before ischemia.In group 5-HD,5-HD 15 mg/kg was injected intraperitoneally at 30 min before ischemia and the other procedures were similar to those previously described in group Ⅰ-pre.Neurological behavior was evaluated at 24 h of reperfusion.The rats in each group were sacrificed at 72 h of reperfusion,and the brains were removed for determination of neuronal apoptosis （by TUNEL） and expression of caspase-3 and phosphor-JNK （p-JNK） protein （using Western blot） in hippocampal tissues.Apoptotic rate was calculated.Results Compared with group S,the number of grid cross was significantly decreased,hanging time was shortened,apoptotic rate was increased,and caspase-3 expression was up-regulated in I/R,Ⅰ-pre and 5-HD groups,the expression of p-JNK protein was up-regulated in IR and 5-HD groups,and no significant change was found in the expression of p-JNK protein in group Ⅰ-pre.Compare with group I/R,the number of grid cross was significantly increased,hanging time was prolonged,apoptotic rate was decreased,and the expression of caspase-3 and p-JNK protein was downregulated in group Ⅰ-pre,and no significant change was found in the parameters mentioned above in group 5-HD.Compared with group Ⅰ-pre,the number of g

关 键 词：KATP通道 JNK丝裂原活化蛋白激酶类 异氟醚 再灌注损伤 脑 缺血预处理 

分 类 号：R742[医药卫生—神经病学与精神病学]