脑穿刺损伤并局部注药大鼠模型的构建初探  

Establishment of Rat Model of Stab Wound Brain Injury and Partly Disemboguing Drug

作　　者：李孝生[1] 刘址忠[2] 陈鹏[1] 谢艳[1] 黄文英[3] 陈家玉[1] 林云东[1] 欧阳兆书[1] 

机构地区：[1]湘南学院附属医院神经外科,湖南郴州423000 [2]南方医科大学南方医院消化科,广州510515 [3]郴州市第一人民医院高压氧科,湖南郴州423000

出　　处：《南昌大学学报（医学版）》2011年第6期1-5,共5页Journal of Nanchang University：Medical Sciences

基　　金：湖南省自然科学基金(09JJ6029);湖南省高等学校科学研究项目[(2008)A065];湖南省医药卫生科研计划项目(B2009122)

摘　　要：目的建立脑穿刺损伤并局部注药大鼠模型,为进一步研究奠定基础。方法成熟的wistar雌性大鼠20只,按随机数字表法分为正常对照组和脑穿刺损伤组,每组10只。脑穿刺损伤组大鼠麻醉后,微型钻钻孔、将针刺入颅内,经顶叶皮层插入脑组织中4 mm,伤及皮层,刺入位置为中线旁开3 mm,额状缝前3 mm,留针2 min,并用X线侧位片和正位片检查刺入位置和深度后退出。正常对照组仅开颅,不致伤。Wistar大鼠脑穿刺后,消毒断头取大鼠皮层细胞原代培养,经纯化后星形细胞传代培养,取第3代星形胶质细胞进行GFPA的免疫组织化学检查鉴定细胞形态,观察脑穿刺损伤组与正常对照组星形胶质细胞的光密度OD值、数密度值。结果经原代及传代培养,获得大量几乎为单一种类细胞,其特点为胞体较大,形状不规则,胞质较丰富,细胞核圆形或卵圆形,常偏于胞体一侧,细胞突起较多较长,符合星形胶质细胞形态。免疫荧光检测显示GFAP阳性细胞为97%,脑穿刺损伤组原代培养细胞数量增多,脑穿刺损伤组与正常对照组星形胶质细胞的数密度分别为73.41±6.53和51.37±5.36,2组比较差异有统计学意义(P<0.05)。脑穿刺损伤组与正常对照组星形胶质细胞的光密度值分别为2.41±0.34和1.37±0.38,2组比较差异有统计学意义(P<0.01)。结论脑穿刺损伤并局部注药大鼠模型可以观察穿刺损伤部位的星形胶质细胞的变化,为在细胞水平研究创伤性脑损伤提供了平台。Objective To establish a rat model of stab wound brain injury and partly disemboguing drug,and to lay the background for further study.Methods Twenty mature female Wistar rats were randomly divided into control group and brain stab trauma group,with 10 rats in each group.In brain stab trauma group,a hole was drilled with a micro-drill after anaesthesia and a needle was inserted 4 mm into the brain in the parietal cortex.The puncture site was settled in the region 3 mm lateral to midline and 3 mm anterior to frontal suture.The needle was retained for 2 minutes,and then was drawed after lateral and anteroposterior X-ray examination for the location and depth of the insertion.Rats in control group underwent craniotomy,but received no injury.After encephalopuncture,Wistar rats were sterilized and decapitated,and cortical cells were obtained and cultured.Following purification,astrocytes were harvested and subcultured.Cultured astrocytes at passage 3 were identified by GFAP immunohistochemistry and the changes in cell morphology were observed with the microscope.The optical density values and numerical density values were compared between the two groups.Results A large number of pure cells were obtained after primary culture and subculture.The cells had the morphological characteristics of astrocytes：large cell body,irregular shape,abundant cytoplasm,many long cellular processes,and round or oval nuclei which biased towards one side of cell body.Immunohistochemical staining demonstrated that 97% of the cells were GFAP positive.The number of cells in primary culture increased in brain stab trauma group.Compared with control group,both optical density values and numerical density values significantly increased in brain stab trauma group（optical density values：2.41±0.34 vs 1.37±0.38,P0.01;numerical density values： 73.41±6.53 vs 51.37±5.36,P0.05）.Conclusion The established rat model of stab wound brain injury and partly disemboguing drug may be used to observe the changes in morphology of astrocytes,and can

关 键 词：脑穿刺损伤 局部注药 模型 原代培养 胶质细胞 动物 实验 大鼠 

分 类 号：R-332[医药卫生]