CAJ | 학술논문

目的探讨七氟醚预处理对大鼠创伤性脑损伤后自噬的影响及c-Jun氨基末端激酶(JNK)信号通路在其中的作用.方法健康雄性成年SD大鼠60只,体重220 ～ 250 g,采用随机数字表法分为4组(n=15):假手术组(S组)、创伤性脑损伤组(TBI组)、创伤性脑损伤+七氟醚预处理组(刨伤性脑损伤+ Sev组)和创伤性脑损伤+七氟醚预处理+JNK选择性拮抗剂SP600125组(TBl+ Sev+ SP组).采用Feeney法建立大鼠创伤性脑损伤模型.TBI+ Sev组和TBI+Sev+SP吸入2.4％七氟醚30 min,1次/d,连续4d,行七氟醚预处理,七氟醚预处理结束后24 h时制备创伤性脑损伤模型.TBI+ Sev+ SP组于脑损伤模型制备后30 min时腹腔注射SP600125 6 mg/kg.于模型制备后1、3、7d随机取5只大鼠进行神经功能缺陷评分,评分结束后,处死大鼠,取脑组织,计算脑水含量,PCR检测脑组织LC3 Ⅱ和Beclin-1的mRNA表达,Western blot检测脑组织LC3Ⅱ蛋白、Beclin-l蛋白、JNK和磷酸化JNK(p-JNK)的表达水平.结果与S组比较,其余3组脑水含量和神经功能缺陷评分升高,脑组织LC3 Ⅱ和Beclin-1的mRNA及其蛋白、JNK和p-JNK表达上调(P＜0.05);与TBI组比较,TBI+ Sev组和TBI+ Sev+ SP组脑水含量和神经功能缺陷评分降低,脑组织LC3 Ⅱ和Beclin-1的mRNA及其蛋白、JNK和p-JNK表达下调(P＜0.05或0.01);与TBI+ Sev组比较,TBI+ Sev+ SP组脑水含量和神经功能缺陷评分降低,脑组织LC3 Ⅱ和Beclin-1的mRNA及其蛋白、JNK和p-JNK表达下调(P＜0.05).结论七氟醚预处理减轻大鼠创伤性脑损伤的机制与抑制脑组织JNK信号通路活化,从而降低自噬有关.
목적 탐토칠불미예처리대대서창상성뇌손상후자서적영향급c-Jun안기말단격매(JNK)신호통로재기중적작용.방법 건강웅성성년SD대서60지,체중220 ～ 250 g,채용수궤수자표법분위4조(n=15):가수술조(S조)、창상성뇌손상조(TBI조)、창상성뇌손상+칠불미예처리조(포상성뇌손상+ Sev조)화창상성뇌손상+칠불미예처리+JNK선택성길항제SP600125조(TBl+ Sev+ SP조).채용Feeney법건립대서창상성뇌손상모형.TBI+ Sev조화TBI+Sev+SP흡입2.4％칠불미30 min,1차/d,련속4d,행칠불미예처리,칠불미예처리결속후24 h시제비창상성뇌손상모형.TBI+ Sev+ SP조우뇌손상모형제비후30 min시복강주사SP600125 6 mg/kg.우모형제비후1、3、7d수궤취5지대서진행신경공능결함평분,평분결속후,처사대서,취뇌조직,계산뇌수함량,PCR검측뇌조직LC3 Ⅱ화Beclin-1적mRNA표체,Western blot검측뇌조직LC3Ⅱ단백、Beclin-l단백、JNK화린산화JNK(p-JNK)적표체수평.결과 여S조비교,기여3조뇌수함량화신경공능결함평분승고,뇌조직LC3 Ⅱ화Beclin-1적mRNA급기단백、JNK화p-JNK표체상조(P＜0.05);여TBI조비교,TBI+ Sev조화TBI+ Sev+ SP조뇌수함량화신경공능결함평분강저,뇌조직LC3 Ⅱ화Beclin-1적mRNA급기단백、JNK화p-JNK표체하조(P＜0.05혹0.01);여TBI+ Sev조비교,TBI+ Sev+ SP조뇌수함량화신경공능결함평분강저,뇌조직LC3 Ⅱ화Beclin-1적mRNA급기단백、JNK화p-JNK표체하조(P＜0.05).결론 칠불미예처리감경대서창상성뇌손상적궤제여억제뇌조직JNK신호통로활화,종이강저자서유관.
Objective To investigate the effects of sevoflurane preconditioning on autophagy after traumatic brain injury (TBI) in rats and the role of C-Jun N-terminal kinase (JNK) signaling pathway.Methods Sixty adult male Sprague-Dawley rats,weighing 220-250 g,were randomly divided into 4 groups (n =15 each) using a random number table:sham operation group (group S),group TBI,TBI + sevoflurane preconditioning group (group TBI + Sevo) and TBI + sevoflurane preconditioning + JNK inhibitor SP600125 group (group TBI + Sev + SP).TBI models were established using Feeney' s method.In TBI + Sev and TBI + Sev + SP groups,the rats inhaled 2.4％ sevoflurane for 30 min once a day for 4 concecutive days,and TBI was produced at 24 h after the end of sevoflurane preconditioning.In TBI + Sev + SP group,SP600125 (6 mg/kg) was injected intrapetitoneally at 30 min after TBI.Five rats were chosen at day 1,3,and 7 after TBI,and neurological deficit score (NDS) was measured.The rats were then sacrificed and brains were removed to measure brain water content,expression of LC3 lⅡ and Beclin-1 mRNA (using PCR),and expression of LC3 Ⅱ,Beclin-1,JNK and phosphorylated JNK (p-JNK) (by Western blot).Results Compared with group S,brain water content and NDS were significantly increased,and the expression of LC3 Ⅱ and Beclin-1 protein and mRNA,JNK,and p-JNK was up-regulated in the other three groups.Brain water content and NDS were significantly decreased,and the expression of LC3 Ⅱ and Beclin-1 protein and mRNA,JNK,and p-JNK was down-regulated in TBI + Sev and TBI + Sev + SP groups as compared with group TBI,and in TBI + Sev + SP group as compared with TBI + Sev group.Conclusion The mechanism by which sevoflurane preconditioning mitigates TBI is related to inhibiton of activation of JNK signaling pathway and decreased autophagy in rats.