他克莫司对创伤性脑损伤大鼠神经行为和记忆能力的影响(英文)

Effect of tacrolimus on neural behaviors and memory of rats with traumatic brain injury

背景:目前他克莫司对周围神经保护作用的研究比较多,而其是否具有促进中枢神经再生和保护作用。目的:通过神经行为学观察和检测突触素的表达及细胞凋亡数量,探讨他克莫司对大鼠脑液压伤后神经行为和记忆的影响。设计:随机分组,平行对照(自我前后对照、相互对照)实验。单位:大连医科大学附属第二医院神经外科和复旦大学附属中山医院神经外科。材料:实验于2002-11在上海长征医院神经外科实验室完成。选择由复旦大学实验动物科学部提供的雄性SD 大鼠24只为观察对象,体质量(200±20)g,用抽签法随机分为对照组、损伤组和治疗组,每组8只。干预:伤前24h 以20g/L 戊巴比妥钠(40m g/kg)腹腔麻醉,损伤组和治疗组模型制备参照美国Dixon 建立的大鼠脑液压损伤模型,打击能量为151.95～172.21kPa,相当于中度颅脑损伤,对照组只埋管不打击。治疗组伤后5m in 以1mg/kg 腹腔注射他克莫司,1次/d,连续7d。损伤组和对照组腹腔注射生理盐水。伤前、伤后3d 和伤后1周分别进行行走试验、平衡试验和记忆试验。于伤后1周全部大鼠断头取脑,分别取海马、伤灶周围额叶皮质、基底核部位进行免疫组织化学检测,图像定量分析技术检测突触素在海马各区以及伤灶周围的额叶皮质与基底节的分布,用流式细胞仪检测凋亡细胞计数。主要观察指标:①各组动物于伤前、伤后3d 和伤后1周时行走试验、平衡试验和记忆试验成绩。②各组动物皮质、海马、基底核区突触素表达定量分析结果。③各组动物皮质、海马、基底核区细胞凋亡百分率。结果:24只大鼠全部进入结果分析。①治疗组伤后3d 和伤后1周时行走试验成绩和平衡试验成绩低于损伤组犤行走试验成绩:(7.5±2.5)s,(5.5±2.1)s,(10.5±2.5)s,(8.2±2.5)s。平衡试验成绩:(3.4±0.5)分,(2.5±0.2)分,(5.7±0.2)分,(5.0±0.5)分,P <0.05～0.01犦,治疗组伤后3d和伤后1周时记忆实验成绩高于损伤组犤(4.9±1.7)s,(6.2±2.3)s,(4.0±1.5)s,(4.4±2.6)s,P <0.05～0.01犦。②治疗组皮质、海马、基底核区突触素表达量显著高于损伤组(140.36±3.87,45.52±2.16,31.67±2.35,96.25±2.85,24.35±2.47,20.49±2.08,P <0.01);对照组显著高于治疗组(162.24±3.52,50.58±2.31,42.69±2.53,P <0.01)。③治疗组海马、皮质、基底节的细胞凋亡率显著低于损伤组犤(10.37±2.12)%,(18.39±2.87)%,(12.78±2.45)%,(21.14±4.85)%,(38.57±3.78)%,(21.18±4.59)%,P <0.01犦。损伤组海马、皮质、基底核的细胞凋亡率显著高于对照组犤(3.85±2.56)%,(4.96±2.15)%,(3.52±2.17)%,P <0.01犦。结论:他克莫司治疗后能促进大鼠海马、皮质及基底核区突触素表达,减少海马和皮质及基底核区神经细胞凋亡,对实验动物脑损伤后神经行为和记忆的恢复有促进作用。

BACKGROUND：At present, many researches have shown that tacrolimus has protecting effect on peripheral nerves,however,whether tacrolimus has a similar effect on central neural system has not been known yet. OBJECTIVE：To investigate the effect of tacrolimus on neural behaviors and memory by observing the neural behaviors and detecting the expression of synaptophysin in cortex,hippocampus and basipodite. DESIGN：Parallel and before and post contrast study. SETTING：Department of neurosurgery, affiliated hospital of the two medicad universities. MATERIALS：The experiment was carried out in the Laboratory of Department Neurosurgery,Shanghai Changzheng Hospital in November 2002. Twenty-four qualified Sprague-Dawley rats supplied by Shanghai Experimental Animal Center of medical institution of Fudan University,with body mass of 180-220 g,were randomly divided into three groups：control, injury and treatment,with 8 rats in each group. INTERVENTIONS：Anesthesia was induced by administration of sodium pentobarbital（with the concentration of 20 g/L,40 mg/kg） twenty-four hours before the operation.The coup energy was 151.95-172.21 kPa that was equal to medium injury.The control rats did not receive the coup.Tacrolimus was given to the treatment group in the dosage of 1 mg/kg body weight by intraperitoneal injection at 5 minutes post trauma.The drug was given once a day till the seventh day post trauma.The injury and control rats received physiological saline.Before and post trauma （on the third and seventh day） all rats received Beam Walking Test,Beam Balance Test and Memory Test, respectively.One week after trauma,all rats＇ heads was removed and the brain was taken out within seconds.The hippocampus,cortex and basipodite of the traumatic hemisphere were kept in methanal with the concentration of 4 percent.Immunohistochemistry and image analysis system were used to study the expression of synaptophysin in cortex,hippocampus and basipodite of the traumatic hemisphere. MAIN OUTCOME MEASURES： ①The performance records of neural behaviors and memory of the three groups. ②Resuhs of the expression of synaptophysin in cortex, hippocampus and basipodite of the three groups. ③Apoptotic cellscalculation of synaptophysin in cortex, hippocampus and basipodite. RESULTS：Totally 24 rats entered the final analysis. ①On the third day and the seventh day,the scores recorded in Walking Test and Balance Test of the treatment group were less than those in the injury group[Walking Test： （7.5±2.5） s,（5.5±2.1） s,（10.5±2.5） s,（8.2±2.5） s.Balance Test：（3.4±0.5） seore,（2.5±0.2） score,（5.7±0.2） score,（5.0±0.5） seore,P 〈 0.05-0.01].But the scores recorded in memory test of the treatment group were still more than those in the control group,memory test[（4.9±1.7） s,（6.2±2.3） s,（4.0±1.5） s, （4.4±2.6） s,P 〈 0.05-0.01]. ②The expression of synaptophysin in cortex, hippocampus and basipodite in the treatment group was still more than that in the injury group（140.36±3.87, 45.52±2.16,31.67±2.35,96.25±2.85, 24.35 ±2.47,20.49±2.08,P 〈 0.01）;The expression of synaptophysin control group was especially more than that in treatment group （162.24±3.52, 50.58±2.31,42.69±2.53, P 〈 0.01）.Apoptotic ceils calculation of cortex,hippocampus and basipodite in treatment group was far less than those in injury group,（10.37 ±2.12）%,（18.39 ±2.87）%,（12.78 ±2.45）%,（21.14 ±4.85）%, （38.57±3.78）%, （21.18±4.59）%,P 〈 0.01].And apoptotic cells calculation im injury group was more than those in control group [（3.85±2.56）%, （4.96±9.15）%, （3.52±2.17）%,P 〈 0.01]. CONCLUSION：Tacrolimus can facilitate the expression of synaptophysin, refrain from calcineurin involving in the process of transcription-dependent and nontranscription-dependent apoptosis,and can also contribute to the recovery of memory and neural behaviors post trauma.