CAJ | 학술논문

目的评价c-Jun氨基末端激酶(Jun N-terminal kinase,JNK)信号通路在异氟烷(isoflurane,ISO)预处理减轻脑缺血再灌注(ischemia/reperfusion,I/R)损伤中的作用,探讨JNK信号通路与细胞凋亡的关系. 方法采用四血管阻断法制作全脑缺血再灌注模型.选择清洁级雄性SD大鼠40只,按随机数字表法将其分为假手术组(S组)、缺血再灌注伤组(I/R组)、JNK信号通路抑制剂SP600125+ I/R组(SP+ I/R组)、ISO预处理组(ISO组)和ISO预处理+JNK信号通路抑制剂SP600125组(ISO+ SP组),每组8只.预处理方法为连续5d吸入15 g/L ISO,1 h/d.末次预处理24h后行I/R,72 h后取脑组织作常规HE染色观察其组织形态学变化,并用TUNEL法检测海马中神经细胞的凋亡率,用免疫组化染色检测caspase-3在海马神经细胞中的表达,采用Western blot法检测海马组织p-JNK蛋白表达. 结果与S组比较,其余各组的脑损伤评分、神经细胞凋亡率、caspase-3表达均升高(P＜0.05),I/R组的p-JNK蛋白表达升高(P＜0.05),而SP+ I/R组、ISO组、ISO+ SP组的p-JNK蛋白表达与S组比较差异无统计学意义(P＞0.05)；与I/R组比较,SP+ I/R组、ISO组、ISO+ SP组的脑损伤评分、神经细胞凋亡率、caspase-3表达、p-JNK蛋白表达均降低(P＜0.05)；与SP+ I/R组和ISO组比较,ISO+ SP组脑损伤评分、神经细胞凋亡率、caspase-3表达进一步降低(P＜0.05),p-JNK蛋白表达差异无统计学意义；SP+ I/R组和ISO组比较各项指标差异无统计学意义. 结论 ISO预处理可通过抑制JNK信号通路、下调caspase-3的表达来减轻脑缺血再灌注损伤.
목적 평개c-Jun안기말단격매(Jun N-terminal kinase,JNK)신호통로재이불완(isoflurane,ISO)예처리감경뇌결혈재관주(ischemia/reperfusion,I/R)손상중적작용,탐토JNK신호통로여세포조망적관계. 방법 채용사혈관조단법제작전뇌결혈재관주모형.선택청길급웅성SD대서40지,안수궤수자표법장기분위가수술조(S조)、결혈재관주상조(I/R조)、JNK신호통로억제제SP600125+ I/R조(SP+ I/R조)、ISO예처리조(ISO조)화ISO예처리+JNK신호통로억제제SP600125조(ISO+ SP조),매조8지.예처리방법위련속5d흡입15 g/L ISO,1 h/d.말차예처리24h후행I/R,72 h후취뇌조직작상규HE염색관찰기조직형태학변화,병용TUNEL법검측해마중신경세포적조망솔,용면역조화염색검측caspase-3재해마신경세포중적표체,채용Western blot법검측해마조직p-JNK단백표체. 결과 여S조비교,기여각조적뇌손상평분、신경세포조망솔、caspase-3표체균승고(P＜0.05),I/R조적p-JNK단백표체승고(P＜0.05),이SP+ I/R조、ISO조、ISO+ SP조적p-JNK단백표체여S조비교차이무통계학의의(P＞0.05)；여I/R조비교,SP+ I/R조、ISO조、ISO+ SP조적뇌손상평분、신경세포조망솔、caspase-3표체、p-JNK단백표체균강저(P＜0.05)；여SP+ I/R조화ISO조비교,ISO+ SP조뇌손상평분、신경세포조망솔、caspase-3표체진일보강저(P＜0.05),p-JNK단백표체차이무통계학의의；SP+ I/R조화ISO조비교각항지표차이무통계학의의. 결론 ISO예처리가통과억제JNK신호통로、하조caspase-3적표체래감경뇌결혈재관주손상.
Objective To evaluate the role of c-Jun N-terminal kinase (JNK) signaling pathway in isoflurane (ISO) preconditioning against cerebral ischemia/reperfusion (I/R) injury and investigate the relationship between JNK signaling pathway and apoptosis.Methods Global cerebral I/R models were made by 4-artery occlusion technique.Forty male SD rats of clean grade were divided into sham operation group (S group),I/R injury group (I/R group),SP600125 (an inhibitor of JNK signaling pathway) + I/R group (SP + I/R group),ISO preconditioning group (ISO group),and ISO preconditioning + SP600125 group (ISO + SP group) according to the random number table.Preconditioning protocol was successive inhalation of 15 g/L ISO for 5 days,1 h/d.I/R was induced at 24 hours after the end of preconditioning.Brain tissues were harvested at 72 hours later to take histomorphological examination by HE staining as well as detect apoptosis of hippocampal nerve cells by TUNEL method,expression of caspase-3 in hippocampal nerve cells by immuno-histochemistry,and expression of protein p-JNK in hippocampal tissues by Western blot.Results Compared with S group,brain injury score,apoptosis ratio of nerve cells,and expression of caspase-3 were significantly increased in the other groups (P ＜ 0.05).Moreover,p-JNK protein had a higher expression in IR group than in S group (P ＜ 0.05),but no significant difference was observed in SP + I/R group,ISO group,and ISO + SP group as compared with S group (P ＞ 0.05).Compared with I/R group,brain injury score,apoptosis ratio of nerve cells,expression of caspase-3,and expression of p-JNK protein were all declined in SP + I/R group,ISO group,and ISO + SP group (P ＜ 0.05).Moreover,brain injury score,apoptosis ratio of nerve cells,and expression of caspase-3 had further decline in ISO + SP group as compared with SP + I/R group and ISO group (P ＜ 0.05),but the difference in expression of p-JNK protein was insignificant among the three groups.Compared with SP + I/R group,no significant changes of each index were found in ISO group.Conclusion ISO preconditioning alleviates cerebral I/R injury through down-regulating expression of caspase-3 and inhibiting JNK signaling pathway.