摘要
目的探讨非阿片类阿片受体激动剂Herkinorin后处理的脑保护作用以及典型蛋白激酶Cγ(conventional protein kinase Cγ,c PKCγ)的作用。方法成年雄性C57BL/6小鼠按数字表法随机分为对照组(Naive),假手术组(Sham),模型组(ischemia/reperfusion,I/R),溶剂组(I/R+D,再灌注前腹腔注射同等剂量的DMSO),Herkinorin组(I/R+H,再灌注前腹腔注射10 mg/kg Herkinorin)。应用小鼠脑中动脉阻塞(middle cerebral artery occlusion,MCAO)诱导缺血性脑卒中模型,通过神经行为和运动功能检测评估脑损伤程度,借助2,3,5-氯化三苯基四氮唑(2,3,5-triph-enyltetrazolium chloride,TTC)染色观察脑梗死体积,蛋白印迹检测c PKCγ膜转位(激活)水平。结果与I/R组比较,I/R+H组缺血再灌注后24 h和7 d的神经行为评分明显降低,爬杆实验、圆柱体实验和步错实验测评明显改善(P<0.05,n=6)。TTC染色显示I/R组梗死体积24 h为31.44%±5.44%,7 d为23.44%±7.95%,I/R+H组(24 h:17.19%±3.23%,7 d:13.26%±2.71%)脑梗死体积明显减少(P<0.05,n=6)。Western blotting结果显示,I/R组缺血核心区(Ic)和半影区(P)中c PKCγ膜转位水平都明显下降,而Herkinorin可明显减轻MCAO小鼠半影区内c PKCγ膜转位水平的降低(P<0.05,n=6)。结论 10 mg/kg Herkinorin腹腔注射能减轻MCAO小鼠皮质的缺血再灌注损伤,c PKCγ膜转位水平的变化可能参与Herkinorin后处理脑保护的分子机制。
Objective To evaluate the effects of Herkinorin postconditioning against ischemia/reperfusion injury in middle cerebral artery occlusion （MCAO） mice and the role of conventional protein kinase Cγ （cPKCγ）. Methods Healthy adult male C57BL/6 mice were randomly divided into five groups：control group （Naive）, sham-operation group （Sham）, ischemia for 1 h and reperfusion group （I/R）, I/R and DMSO intraperitoneal injection group （I/R＋D）,I/R and 10 mg/kg Herkinorin intraperitoneal injection group （I/R＋H）. Neurobehavioral score, Pole test, Wire hang test, Cylinder test and Foot fault test were performed at 24 h and 7 d after reperfusion. Using MCAO induced ischemic stroke mouse model, cerebral infarct volume was evaluated with TTC staining, the cPKCγ membrane translocation levels were detected with Western blotting. Results MCAO induced ischemia/reperfusion injury resulted in increased neurobehavioral scores in mice. Compared with I/R group, neurobehavioral scores of I/R＋H group were decreased significantly. Ischemia/reperfusion injury affected sensorimotor function. Pole test, Cylinder test and Foot fault test of I/R＋H group were relieved significantly compared with I/R group （P〈0.05,n=6）. TTC staining showed that infarct volumes in I/R group were 31.44%±5.44% at 24 h and 23.44%±7.95% at 7 d, infarct volumes were decreased in I/R＋H group （24 h：17.19%±3.23%,7 d：13.26%±2.71%） （P〈0.05,n=6）. Western blotting showed cPKCγ membrane translocation levels of ischemic core and peri-infarct regions in MCAO mice cortex were decreased significantly after ischemia for 1 h and reperfusion. Herkinorin alleviated the decrease of cPKCγ membrane translocation levels in cortex peri-infarct region （P〈0.05,n=6）. Conclusion 10 mg/kg Herkinorin could decrease neurobehavioral score, alleviate Pole test, Cylinder test and Foot fault test evaluation, decrease infarct volumes in MCAO mice. cPKCγ membrane translocation in cortex peri-infarct region may participate molecular
出处
《首都医科大学学报》
CAS
北大核心
2017年第3期336-341,共6页
Journal of Capital Medical University
基金
国家自然科学基金(81301065)
北京市优秀人才培养资助计划(D003034000031)
首都医科大学附属北京同仁医院科研骨干培育基金(2014-YJJ-GGL-044)~~
