CAJ | 학술논문

目的评价异氟醚后处理对大鼠脑缺血再灌注损伤的影响.方法健康雄性SD大鼠32只,体重280～320 g,采用随机数字表法,将其分为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)和异氟醚后处理组(Ⅰ-post组).采用改良Pulsinelli四血管阻断法制备大鼠全脑缺血再灌注损伤模型.Ⅰ-pre组缺血前吸入1.5％异氟醚2h,Ⅰ-post组再灌注即刻吸入1.5％异氟醚30 min.于再灌注24h时行神经行为学评分,再灌注72h时处死大鼠取海马,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,Western blot法检测磷酸化c-Jun氨基末端激酶(p-JNK)表达.结果与S组比较,I/R组、Ⅰ-pre组和Ⅰ-post组格子爬行数减少,转体时间延长,悬挂时间缩短,海马神经元凋亡率升高,caspase-3和p-JNK表达上调(P＜0.05)；与I/R组比较,Ⅰ-pre组和Ⅰ-post组格子爬行数增多,转体时间缩短,悬挂时间延长,海马神经元凋亡率降低,caspase-3和p-JNK表达下调(P＜0.05)；与Ⅰ-pre组比较,Ⅰ-post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(P＜0.05),p-JNK表达差异无统计学意义(P＞0.05).结论异氟醚后处理可减轻大鼠脑缺血再灌注损伤,且其效果弱于预处理,其机制与激活海马JNK信号转导通路抑制神经元凋亡有关.
목적 평개이불미후처리대대서뇌결혈재관주손상적영향.방법 건강웅성SD대서32지,체중280～320 g,채용수궤수자표법,장기분위4조(n=8):가수술조(S조)、뇌결혈재관주조(I/R조)、이불미예처리조(Ⅰ-pre조)화이불미후처리조(Ⅰ-post조).채용개량Pulsinelli사혈관조단법제비대서전뇌결혈재관주손상모형.Ⅰ-pre조결혈전흡입1.5％이불미2h,Ⅰ-post조재관주즉각흡입1.5％이불미30 min.우재관주24h시행신경행위학평분,재관주72h시처사대서취해마,채용TUNEL법검측조망신경원,계산신경원조망솔,면역조화법검측caspase-3표체,Western blot법검측린산화c-Jun안기말단격매(p-JNK)표체.결과 여S조비교,I/R조、Ⅰ-pre조화Ⅰ-post조격자파행수감소,전체시간연장,현괘시간축단,해마신경원조망솔승고,caspase-3화p-JNK표체상조(P＜0.05)；여I/R조비교,Ⅰ-pre조화Ⅰ-post조격자파행수증다,전체시간축단,현괘시간연장,해마신경원조망솔강저,caspase-3화p-JNK표체하조(P＜0.05)；여Ⅰ-pre조비교,Ⅰ-post조격자파행수감소,현괘시간축단,해마신경원조망솔승고,caspase-3표체상조(P＜0.05),p-JNK표체차이무통계학의의(P＞0.05).결론 이불미후처리가감경대서뇌결혈재관주손상,차기효과약우예처리,기궤제여격활해마JNK신호전도통로억제신경원조망유관.
Objective To evaluate the effects of isoflurane postconditioning on cerebral ischemia-reperfusion (I/R) injury in rats.Methods Thirty-two male Sprague-Dawley rats,weighing 280-320 g,were randomly divided into 4 groups (n=8 each):group sham operation (group S),group I/R,group isoflurane preconditioning (group Ⅰ-pre),and group isoflurane postconditioning (group Ⅰ-post).Global cerebral I/R was induced by 4-vessel occlusion method described by Pulsinelli.1.5％ isoflurane was inhaled for 2h before ischemia in group Ⅰ-pre.1.5％ isoflurane was inhaled for 30 min immediately after onset of reperfusion in group Ⅰ-post.Neurological function was assessed and scored at 24h of reperfusion.The rats were sacrificed at 72h of reperfusion and hippocampi were isolated for determination of neuronal apoptosis (by TUNEL),caspase-3 expression (by immuno-histochemistry),and phosphorylated c-Jun N-terminal kinase (p-JNK) protein expression (by using Western blot) in hippocampal tissues.Apoptotic rate was calculated.Results Compared with S group,the number of grid cross was decreased,twist time was prolonged,hanging time was shortened,apoptotic rate was increased,and the expression of caspase-3 and p-JNK protein was up-regulated in I/R,I-pre and Ⅰ-post groups (P ＜ 0.05).Compared in I/R group,the number of grid cross was increased,twist time was shortened,hanging time was prolonged,apoptotic rate was decreased,and the expression of caspase-3 and p-JNK protein was down-regulated in Ⅰ-pre and Ⅰ-post groups (P ＜ 0.05).Compared with I/R group,the number of grid cross was decreased,hanging time was shortened,apoptotic rate was increased,and the expression of caspase-3 was up-regulated (P ＜ 0.05),and no significant changes in the expression of p-JNK protein were found in Ⅰ-post group (P ＞ 0.05).Conclusion Isoflurane postconditioning can reduce cerebral I/R injury,the efficacy is weaker than that of preconditioning and the mechanism is related to activation of JNK signal transduction pathway and inhibition of neuronal apoptosis in hippocampus of rats.