器官型脑片培养OGD复氧复糖模型中丁苯酞对于bcl-2/bax、及bim蛋白的调节作用  

The effect of NBP on bcl-2 / bax and bim proteins in oxygen-glucose deprivation model of organotypic brain slices

作　　者：董梅[1] 程树彬[2] 郭彦芳 李钊[4] 李世平[1] 张祥建[1] 李春岩[1] DONG Mei CHENG Shubin GUO Yanfang et al(Department of Neurology, Key Laboratory of Neurology of Hebei Province, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, Chin)

机构地区：[1]河北医科大学第二医院神经内科,神经病学河北省重点实验室,河北石家庄050000 [2]河北工程大学附属医院神经内科,河北邯郸056002 [3]邢台市一院神经内科,河北邢台059001 [4]河北医科大学第三医院神经外科,河北石家庄050051

出　　处：《中风与神经疾病杂志》2017年第1期16-20,共5页Journal of Apoplexy and Nervous Diseases

摘　　要：目的观察正丁基苯酞(NBP)对乳大鼠大脑皮质器官型脑片的氧糖剥夺模型(OGD)中细胞凋亡bcl-2、bax及bim蛋白的调节作用。方法用生后7 d的SD乳大鼠制备大脑皮质运动区器官型脑片,将培养2 w的脑片随机分为对照组(C组)、模型组(M组)、溶剂对照组(Sc组)和实验组(T组),T组在OGD干预30 min后给予NBP 10μM。将脑片制备OGD模型30 min,部分脑片在NBP干预前后不同时间点(0 h、1 h、24 h、72 h)进行PI染色;部分脑片在NBP干预后6 h用戊二醛固定,做石蜡切片,行HE染色及bcl-2、bax、bim免疫组化染色。结果 PI染色可见C组在不同时间点荧光强度无明显变化,M组、Sc组和T组荧光强度均随时间有不同程度增强,M组和Sc组各时间点荧光强度无明显差异,T组荧光强度较前两者降低,随时间延长,降低幅度增大,尤其是72 h时间点。HE染色C组可见皮质运动区特有的大锥体细胞,结构清晰,M组和T组可见细胞肿胀,细胞核溶解,M组相对明显。免疫组化染色可见在3种染色中M组和T组阳性细胞数均较C组增多,其中bcl-2染色中T组阳性细胞增多更明显,bax和bim染色中M组阳性细胞增多更明显,3种染色中C、M、T各组之间差异均有统计学意义。结论NBP对OGD后的细胞损伤具有保护作用,可能是通过对bcl-2、bax以及bim蛋白的调节减少细胞凋亡。Objective To establish the oxygen and glucose deprivation（ OGD） model of organotypic brain slices of rat cerebral cortex and observe the effects of n-butyl phthalide（ NBP） on bcl-2、bax、bim protein in apoptosis. Method We chose the post-natal 7-day rats（ SD） to culture the brain slices on the membrane insert and they were divided into four groups after having been cultured for two weeks： control group（ group C）,model group（ group M）,the solvent control group（ group Sc） and test group（ group T）. Group T were given NBP 10μM to brain slices after treating with OGD30 min. Some brain slices were observed by PI dyeing before and after being gived NBP 1 h,24 h and 72 h. Other brain slices were fixed by glutaral after 6 h treated with NBP and were dying by HE and assessed the positive cells of bcl-2、bax、bim in brain slices of different groups. Results Brain slices showed no different brightness of red fluorescence in group C and showed increased brightness according to different time in group M,group Sc and group T by PI staining. There were no variety in group M and group Sc. But the brightness was dicreased in group T than that of group M especially in 72 h（ P〈0. 05）.There were typital giant pyramidal cells in cortical motor area with normal structures. There were much more swelling cells with karyolysis in group M than that in group T. There were more positive cells in group M and group T than that in group C. Among them there were more positive cells of bcl-2 in group T and more positive cells of bax and bim in group M. There were statistic variance in three groups with each dying of bcl-2,bax and bim. Conclusion NBP had protective effect on damaged brain slice after OGD. NBP could decrease apoptosis by controlling the protein of bcl-2,bax and bim.

关 键 词：器官型脑片 OGD 丁苯酞 细胞凋亡 bcl-2 BAX bim 

分 类 号：R965[医药卫生—药理学]