过表达B细胞淋巴瘤/白血病-2对重度创伤性脑损伤大鼠的治疗作用及其机制  

Therapeutic effect of overexpression of B cell lymphoma/leukemia-2 for severe traumatic brain injury in rats and its mechanism

作　　者：张辉[1] 杨林[1] 陈培莉[1] 杨帆[2] Zhang Hui;Yang Lin;Chen Peili;Yang Fan(Department of Critical Care Medicine,Shangqiu First People′s Hospital and Xuzhou Medical University Shangqiu Clinical College,Shangqiu 476100,China;Department of Critical Care Medicine,Neurosurgery,Henan Provincial People′s Hospital,Zhengzhou 450000,China)

机构地区：[1]商丘市第一人民医院重症医学科,徐州医科大学商丘临床学院,476100 [2]河南省人民医院神经外科重症医学科,450000

出　　处：《中华实验外科杂志》2021年第1期92-95,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察过表达B细胞淋巴瘤/白血病-2(bcl-2)对重度创伤性脑损伤大鼠神经保护作用。方法2018年1月至2019年9月,60只SD大鼠采用随机数字法分为对照组、模型组和bcl-2组。对照组和模型组大鼠经脑室注射对照腺相关病毒1×10^11 v.g/只,bcl-2组经脑室注射bcl-2过表达腺相关病毒1×10^11 v.g/只,注射24 d后,模型组和bcl-2组大鼠采用自由落体撞击法建立重度创伤性脑损伤模型,对照组大鼠不处理。建模24 h后,采用神经行为学评分评价大鼠的神经功能;采用原位缺口末端标记法(TUNEL)染色分析脑组织的凋亡水平;采用JQ1法检测脑组织细胞线粒体膜电位;采用蛋白质印迹法(Western blot)分析bcl-2、bcl-2相关X蛋白(bax)和半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3蛋白表达水平,计量资料比较采用单因素方差分析。结果bcl-2组大鼠神经功能评分(5.72±1.27)低于模型组大鼠神经功能评分(8.59±1.67),差异有统计学意义(t=3.114,P<0.05)。bcl-2组大鼠脑组织细胞线粒体膜电位(1.59±0.13)高于模型组大鼠脑组织细胞线粒体膜电位(1.01±0.10),差异有统计学意义(t=2.011,P<0.05)。bcl-2组大鼠神经细胞凋亡水平[(16.59±3.09)%]显著低于模型组[(39.54±4.09)%],差异有统计学意义(t=3.103,P<0.05)。bcl-2组细胞bcl-2蛋白表达水平(1.79±0.14)高于对照组和模型组(0.96±0.11和1.00±0.12),差异有统计学意义(t=2.019、1.978,P<0.05)。bcl-2组大鼠脑组织Caspase-3和bax蛋白表达水平(1.17±0.09和1.05±0.17)低于模型组(1.32±0.11和1.74±0.12),差异有统计学意义(t=1.280、2.016,P<0.05)。结论过表达bcl-2可显著抑制重度创伤性脑损伤大鼠神经细胞凋亡,提高线粒体功能,保护大鼠神经功能。Objective To investigate the neuroprotective effect of overexpression of B cell lymphoma/leukemia-2(bcl-2)on rats with severe traumatic brain injury.Methods From January 2018 to September 2019,a total of 60 SD rats were purchased from Shanghai SLAC Laboratory Animal Co.,Ltd and randomly divided into control group,model group and bcl-2 group.Rats in control group and model group were intraventricularly injected with 1×10^11 v.g control adeno-associated virus,and those in bcl-2 group were injected with 1×10^11 v.g bcl-2.After 24 days of injection,severe traumatic brain injury model was established in model group and bcl-2 group by free falling impact method,while the animals in control group were not treated.At 24 h after modeling,the neurological function of rats was evaluated by neurobehavioral score.The apoptosis level of brain tissue was analyzed by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining.The mitochondrial membrane potential of brain tissue was detected by JC-1 method.The protein expression levels of bcl-2,bcl-2 associated X protein(bax)and cysteinyl aspartate specific proteinase-3(Caspase-3)were analyzed by Western blotting.One-way analysis of variance(ANOVA)was used to compare the data between groups.Results Compared with the model group(8.59±1.67),the neurological function score in bcl-2 group was significantly decreased(5.72±1.27,t=3.114,P<0.05).Compared with the model group(1.01±0.10),the mitochondrial membrane potential(1.59±0.13)in bcl-2 group was significantly increased(t=2.011,P<0.05).Compared with the model group[(39.54±4.09)%],the apoptosis level in bcl-2 group[(16.59±3.09)%]was significantly decreased(t=3.103,P<0.05).Compared with control group and model group[(0.96±0.11)and(1.00±0.12)],the expression level of bcl-2 protein in bcl-2 group was significantly increased(1.79±0.14,t=2.019,1.978,P<0.05).Compared with the expression levels of bax and Caspase-3 protein in the model group(1.32±0.11,1.74±0.12),the expression levels of Caspase-3 and bax prot

关 键 词：B细胞淋巴瘤/白血病-2 脑损伤 凋亡 神经功能 

分 类 号：R651.15[医药卫生—外科学]