大鼠痉挛性脑损伤模型的建立  

Establishment of spastic brain injury model in rats

作　　者：周彤 步玮[3] 张旭[1] 于晓飞[1] 白延彬[1] 李洪杰[1] 曹冉 于亚东[1] Zhou Tong;Bu Wei;Zhang Xu;Yu Xiaofei;Bai Yanbin;Li Hongjie;Cao Ran;Yu Yadong(Department of Hand Surgery,The Third Hospital of Hebei Medical University,Shijiazhuang 050051,China;Department of Hand Surgery,The Second Hospital of Tangshan,Tangshan 063000,China;Department of Neurosurgery,The Third Hospital of Hebei Medical University,Shijiazhuang 050051,China;Department of Orthopaedics,Shijiazhuang People's Hospital,Shijiazhuang 050036,China)

机构地区：[1]河北医科大学第三医院手外科,石家庄050051 [2]河北省唐山市第二医院手外科,唐山063000 [3]河北医科大学第三医院神经外科,石家庄050051 [4]河北省石家庄市人民医院骨外科,石家庄050036

出　　处：《中华解剖与临床杂志》2022年第3期195-200,共6页Chinese Journal of Anatomy and Clinics

基　　金：河北省财政厅优秀人才支撑项目(361005)。

摘　　要：目的探讨无水乙醇内囊靶向注射建立大鼠痉挛性脑损伤模型的可行性。方法选择无特定病原体级雄性SD大鼠60只,鼠龄2~3月龄,体质量290 g~320(304.2±11.6)g。按数字表法随机分为无水乙醇组(ETH组)、生理盐水对照组(NS组)和空白对照组(CK组),每组20只。参照大鼠脑立体定位图谱,使用脑立体定位仪确定大鼠右侧内囊位置后,ETH组注射80μL无水乙醇,NS组注射80μl生理盐水,CK组不注射任何试剂。术后观察各组大鼠左侧上下肢痉挛状态的开始时间、持续时间;术后第3、7、14、21天采用神经系统安全性评分(NSS)标准评估大鼠脑损伤程度,Faden评分标准评估大鼠运动损伤状态,改良Ashworth评分标准评价肌肉痉挛状态。术后第21天取各组大鼠脑组织制备切片,HE染色观察内囊损伤情况;取左侧上肢指总屈肌、左侧下肢腓肠肌组织,免疫荧光法检测Ⅰ型肌纤维百分比。结果术后3天内,ETH组大鼠死亡5只,NS组死亡2只,CK组死亡1只。ETH组大鼠术后第1天即出现左侧上、下肢痉挛状态,症状持续直至术后第21天处死前;NS组、CK组大鼠均无左侧上、下肢痉挛症状。NS组、CK组大鼠术后第3、7、14、21天NSS、改良Ashworth评分均为0分,Faden评分均为22分;ETH组第3、7、14、21天NSS评分分别为(15.5±1.9)、(15.8±1.8)、(15.4±1.7)、(15.1±1.8)分,Faden评分分别为(3.5±1.8)、(3.2±1.7)、(3.7±1.9)、(3.9±1.8)分,改良Ashworth评分分别为(3.5±0.5)、(3.5±0.5)、(2.9±0.7)、(2.9±0.6)分。术后第21天脑组织形态学检测提示:ETH组右侧内囊部位空洞样坏死,神经细胞显著减少,神经细胞结构松散,排列紊乱,细胞核固缩、消失,未累及其他脑区;NS组、CK组脑部神经细胞结构完整,排列有序,细胞核无扭曲、变形,核仁清楚。免疫荧光检测结果显示,ETH组、NS组、CK组左侧上肢指总屈肌和左侧下肢腓肠肌Ⅰ型肌纤维百分比分别为10.2%±6.3%、6.5%±2.9%、6.7%±2.9%和13Objective To explore a method of establishing a rat spastic brain injury model.Methods Sixty specified pathogen-free grade male SD rats aged 2-3 months,with a body mass of 290-320(304.2±11.6)g,were divided into the ethanol group(ETH group,n=20),normal saline group(NS group,n=20),and blank control group(CK group,n=20)by random number method.According to the stereotaxic map of the rat brain,the location of the right internal capsule of the rat was determined by the brain stereotaxic instrument.Then,we injected 80μL of ethanol into the right internal capsule in the ETH group.Analogously,80μL of normal saline and no reagent were injected into the same target in the NS group and CK group,respectively.The general conditions of the rats in the three groups were observed after operation.The onset time and duration of symptoms and the mortality were recorded.On the 3rd,7th,14th,and 21st day after operation,the degree of brain injury was evaluated by neurological severity scores(NSS).The state of motor injury was evaluated by Faden scores,and muscle spasm was evaluated by modified Ashworth scores.On the 21st day after operation,the brain tissues of rats in each group were taken to prepare sections,and HE staining was used to observe the injury of the internal capsule.The total flexor muscle of the left forelimb and the gastrocnemius muscle of the left hindlimb were taken to prepare sections,and the percentage of type Ⅰ muscle fibers was detected by immunofluorescence.Results Within 3 days after surgery,5 rats in ETH group,2 in NS group and 1 in CK group died.The rats in the ETH group showed spasm in the left upper and lower limbs one day after operation,and the symptoms lasted for 21 days.However,no spasm of the left upper and lower limbs was observed in the NS and CK group.The NSS and modified Ashworth scores were all 0 in the NS and CK groups on the 3rd,7th,14th,and 21st day after operation,while their Faden scores were 22 points.In the ETH group,their NSS scores were(15.5±1.9),(15.8±1.8),(15.4±1.7),and(15.1±1.8

关 键 词：脑损伤 脑性瘫痪 痉挛 模型 动物 无水乙醇 内囊 大鼠 

分 类 号：R651.15[医药卫生—外科学] R-332[医药卫生—临床医学]