CAJ | 학술논문

目的探讨异氟烷治疗对大鼠脑外伤后继发性脑损伤的保护作用及机制.方法采用改良Feeney自由落体法制作大鼠创伤性脑损伤模型,将80只雄性SD大鼠按随机数字法分为4组:假手术组、脑外伤组、异氟烷预处理组(脑外伤前12 h给予2％的异氟烷60 min处理)和异氟烷后处理组(脑外伤后10 min中给予2％异氟烷处理60 min),伤后12 h和24 h行神经功能评分,Western blot法检测脑外伤后24 h大脑伤灶周围皮层中磷酸化蛋白激酶B(p-Akt)、磷酸化糖原合成酶激酶3β(p-GSK3β)、活化半胱氨酰天冬氨酸特异性蛋白酶(cleaved-Caspase-3)和B细胞淋巴瘤/白血病-2(bcl-2)和bcl-2相关X蛋白(bax)的表达,原位缺口末端标记法(TUNEL)检测神经细胞的凋亡.结果伤后12 h和24h,与脑外伤组(15.03 ±-3.98、15.12±4.01)比较,异氟烷预处理组(12.58±3.41、12.39±3.13)和异氟烷后处理(12.67±3.07、12.51±3.31)组中大鼠的神经功能缺损评分显著降低,差异有统计学意义(P＜0.05).与脑外伤组比较,异氟烷预处理组和异氟烷后处理组均可以显著上调p-Akt(0.11±0.07、0.98±0.13、1.01±0.17)、p-GSK3β(0.18±0.06、0.56±0.19、0.61±0.18)和bcl-2(0.13±0.05、0.41±0.13、0.49士0.15)的表达,差异有统计学意义(P＜0.05);与脑外伤组(0.78±0.18)比较,脑外伤后伤灶周围脑组织中的cleaved-Caspas-3在异氟烷预处理组(0.32±0.12)和异氟烷后处理组(0.21±0.09)被显著抑制,差异有统计学意义(P＜0.05);4组TUNEL阳性细胞个数比较,差异有统计学意义(F=7.092,P＜0.05),异氟烷预处理组和异氟烷后处理组治疗可显著减少凋亡神经细胞的个数.结论异氟烷可能通过激活Akt/GSK3β信号通路从而在脑外伤中发挥神经保护作用.
목적 탐토이불완치료대대서뇌외상후계발성뇌손상적보호작용급궤제.방법 채용개량Feeney자유락체법제작대서창상성뇌손상모형,장80지웅성SD대서안수궤수자법분위4조:가수술조、뇌외상조、이불완예처리조(뇌외상전12 h급여2％적이불완60 min처리)화이불완후처리조(뇌외상후10 min중급여2％이불완처리60 min),상후12 h화24 h행신경공능평분,Western blot법검측뇌외상후24 h대뇌상조주위피층중린산화단백격매B(p-Akt)、린산화당원합성매격매3β(p-GSK3β)、활화반광안선천동안산특이성단백매(cleaved-Caspase-3)화B세포림파류/백혈병-2(bcl-2)화bcl-2상관X단백(bax)적표체,원위결구말단표기법(TUNEL)검측신경세포적조망.결과 상후12 h화24h,여뇌외상조(15.03 ±-3.98、15.12±4.01)비교,이불완예처리조(12.58±3.41、12.39±3.13)화이불완후처리(12.67±3.07、12.51±3.31)조중대서적신경공능결손평분현저강저,차이유통계학의의(P＜0.05).여뇌외상조비교,이불완예처리조화이불완후처리조균가이현저상조p-Akt(0.11±0.07、0.98±0.13、1.01±0.17)、p-GSK3β(0.18±0.06、0.56±0.19、0.61±0.18)화bcl-2(0.13±0.05、0.41±0.13、0.49사0.15)적표체,차이유통계학의의(P＜0.05);여뇌외상조(0.78±0.18)비교,뇌외상후상조주위뇌조직중적cleaved-Caspas-3재이불완예처리조(0.32±0.12)화이불완후처리조(0.21±0.09)피현저억제,차이유통계학의의(P＜0.05);4조TUNEL양성세포개수비교,차이유통계학의의(F=7.092,P＜0.05),이불완예처리조화이불완후처리조치료가현저감소조망신경세포적개수.결론 이불완가능통과격활Akt/GSK3β신호통로종이재뇌외상중발휘신경보호작용.
Objective To evaluate the neuro-protective effects and mechanism of isoflurane therapy on secondary brain injury in rats with traumatic brain injury (TBI).Methods The TBI model was established according to Feeney' s method.Eighty rats were randomly assigned to four groups : sham group, traumatic brain injury group, isoflurane preconditioning group and isoflurane postprocessing group.Neurological Severity Scores was performed at 12 and 24 h post TBI.Western blotting was used to examine the expression of phosphorylated protein kinase B (p-Akt), phosphorylated glycogen synthase kinase 3β (p-GSK3β), cleaved-cysteinyl aspartate-specific protease (Caspase)-3, B cell lymphoma/leukemia-2 (bcl-2) and bcl-2 associated X protein (bax) in the injured cerebral cortex 24 h post TBI.TdT-mediated dUTP nick end labeling (TUNEL) method was used to detect the apoptosis of nerve cells at the same time.Results Compared with traumatic brain injury group, the Neurological Severity Scores of isoflurane preconditioning and postprocessing group were decreased at 12 and 24 hours post TBI (15.03 ± 3.98, 15.12 ±4.01;12.58 ±3.41, 12.39 ±3.13;12.67 ±3.07, 12.51 ±3.31, P＜0.05).Isoflurane preconditioning and postprocessing could significantly increase the expression of and the expression of p-Akt (0.11 ±0.07, 0.98±0.13, 1.01 ±0.17), p-GSK3β (0.18±0.06, 0.56±0.19, 0.61 ±0.18) and reduce the expression of cleaved-caspas-3 (0.78 ±0.18, 0.32 ±0.12, 0.21 ±0.09) at 24 hours post TBI (P ＜ 0.05).Furthermore, isoflurane preconditioning and postprocessing treatment significantly attenuated neurological deficits and reduced apoptosis after TBI (P ＜ 0.05).Conclusion Isoflurane therapy could exert neuroprotective effect in rats with TBI through stimulating the Akt/GSK3β signaling pathway.