重组质粒pCDNA3.1(+)/Ngb对局灶性脑缺血大鼠脑保护作用的研究  

Protective role of recombinant plasmid pCDNA3.1(+)/Ngb during focal cerebral ischemia of rat brain

作　　者：朱继[1] 唐文渊[1] 

机构地区：[1]重庆医科大学附属第一医院神经外科,重庆400016

出　　处：《第三军医大学学报》2007年第11期1024-1027,共4页Journal of Third Military Medical University

基　　金：重庆市卫生局资助项目(06-2-177)~~

摘　　要：目的研究重组神经球蛋白表达质粒pCDNA3．1（＋）／Ngb对缺血中的脑保护作用。方法54只雄性大鼠按随机数字表法分为生理盐水组、空质粒组和重组神经球蛋白组，每组18只分别于皮质区两部位注射生理盐水、质粒pCDNA3．1（＋）和重组质粒pCDNA3．1（＋）／Ngb，并于注射后24h采用改良线栓法制备大鼠大脑中动脉缺血24h模型。采用TTC染色、原位细胞凋亡检测、间接免疫荧光法和Western blot检测各组大鼠脑梗死面积，神经细胞凋亡状况和bcl-2蛋白表达变化。结果经重组质粒pCDNA3．1（＋）／Ngb处理的大鼠，其脑梗死面积和注射处缺血半暗带凋亡细胞数较其他两组显著减少（P〈0．01），注射处缺血半暗带bcl-2阳性细胞面积百分比显著增高（P〈0．01），bcl-2蛋白的表达上升约40％-50％。结论质粒pCDNA3．1（＋）介导大鼠神经球蛋白基因转入脑内可通过上调bcl-2蛋白表达抑制局灶性脑缺血神经细胞凋亡，起到局灶性脑缺血脑保护作用。Objective To observe the protective effect of recombinant plasmid pCDNA3.1 （ ＋ ）/Ngb during focal cerebral ischemia in rat brain. Methods Fifty-four male Wistar rats were randomly divided into three groups： normal saline（NS） control group, plasmid control group,and recombinant neuroglobulin group [ pCDNA3.1 （ ＋ ）/Ngb]. NS, plasmid pCDNA3.1 （ ＋ ） and recombinant plasmid pCDNA3.1 （ ＋ ）/Ngb were respectively injected into two sites of the rat cerebral cortex 24 hours before induction of neocortical focal ischemia by occlusion of the right middle cerebral artery for 24 hours. The condition of local ischemic damage, expression of bcl-2 and the apoptosis in neural cells were confirmed by staining with 2% 2.3.5-triphenyltetrazolium chloride,in-site cell apoptosis detection, indirect immunofluorescent staining and Western blotting, respectively. Results The extent of cerebral infarction tissue and the apoptosis cells in the pCDNA3.1 （ ＋ ）/Ngb group were significantly reduced than those in other control groups （ P 〈 0.01 ）. The number of expression of bcl-2 cells in pCDNA3.1 （ ＋ ）/Ngb group was much more than that in other control groups （P 〈0.01 ）. The relative expression level of bcl-2 protein was elevated by 40% -50%. Conclusion The neural cells could be protected by recombinant plasmid pCDNA3.1 （ ＋ ）/Ngb during focal cerebral ischemia through up-regulating the expression of bcl-2 gene and retarding the apoptosis of neural cells.

关 键 词：神经球蛋白 脑缺血 神经细胞 BCL-2蛋白 

分 类 号：R394.6[医药卫生—医学遗传学] R741.05[医药卫生—基础医学]