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 cPKCγmembrane 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％ at24 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 cPKCγmembrane translocation levels of ischemic core and periinfarct 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.cPKCγmembrane translocation in cortex peri-infarct region may participate molecular mechanism of Herkinorin induced neuroprotection.%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 proteinkinase Cγ,cPKCγ)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/kgHerkinorin).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-enyhetrazolium chloride,TTC)染色观察脑梗死体积,蛋白印迹检测cPKCγ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7d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).TrC染色显示I/R组梗死体积24h为31.44％ ±5.44％,7d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ePKCγ膜转位水平都明显下降,而Herkinorin可明显减轻MCAO小鼠半影区内cPKCγ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,cPKCγ膜转位水平的变化可能参与Herkinorin后处理脑保护的分子机制.
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