移植神经生长因子和脑衍生神经生长因子基因修饰的嗅鞘细胞对脊髓损伤后细胞凋亡的影响  

Transplantation of nerve growth factor and olfactory ensheathing cells modified by brain-derived nerve growth factor gene in cell apoptosis following spinal cord injury

作　　者：布林[1] 郑鸿[1] 姜汉国[2] 袁即山[3] 

机构地区：[1]广东医学院附属医院骨科,广东省湛江市524001 [2]广东医学院病理教研室,广东省湛江市524001 [3]中山大学第一附属医院烧伤科,广东省广州市510060

出　　处：《中国临床康复》2005年第18期124-125,共2页Chinese Journal of Clinical Rehabilitation

基　　金：广东省卫生厅科研基金课题(A2003552)~~

摘　　要：目的:探讨大鼠脊髓损伤后神经生长因子和脑衍生神经生长因子基因修饰的嗅神经鞘细胞移植对脊髓损伤后细胞凋亡及促进神经功能恢复的作用。方法:实验于2003-09/2004-04在广东医学院实验动物中心完成,SD大鼠48只,雌雄不限,体质量240～260g。随机分为3组:基因修饰组,嗅鞘细胞移植组,脊髓损伤组,每组16只。均首先行大鼠脊髓损伤造模。基因修饰组和嗅鞘细胞移植组分别移植神经生长因子、脑衍生神经生长因子修饰的嗅鞘细胞和单纯嗅鞘细胞,脊髓损伤组不做治疗。移植后12周,采用免疫组织化学法和原位末端标记法对损伤区的脊髓组织切片进行细胞凋亡的检测,主要观察bcl-2(细胞凋亡抑制基因),bax和fas(细胞凋亡诱导基因)阳性表达的情况评估经基因修饰和未经基因修饰的单纯嗅鞘细胞对脊髓神经凋亡的抑制及诱导作用。结果:48只大鼠均进入结果分析。①原位末端标记法检测结果:基因修饰组细胞凋亡数低于嗅鞘细胞移植组,脊髓损伤组最高。②免疫组织化学结果:Bcl-2蛋白表达的顺序为基因修饰组>嗅鞘细胞移植组>脊髓损伤组(20.79±6.40,13.54±3.66,9.34±2.12,P<0.05);Fas,Bax蛋白表达的顺序均为脊髓损伤组>嗅鞘细胞移植组>基因修饰组[(24.98±4.32,40.48±2.05),(18.92±4.74,25.54±3.82),(11.78±3.81,16.87±3.30),(P<0.05)]。结论:单纯移植嗅鞘细胞其存活时间与分泌有限。神经生长因子和脑衍生神经生长因子是神经营养因子的主要成员,用神经生长因子和脑衍生神经生长因子基因修饰嗅鞘细胞后可提高嗅鞘细胞的生存时间和提高分泌神经营养因子功能,移植后具有抑神经细胞凋亡的作用。AIM:To study the effect of intra- cord transplantation of nerve growth facto r(NGF) and brain- derived neurotrophic factor(BDNF) genes modified olfactory en sheathing cells(OECs) on apoptosis after spinal cord injury(SCI) and in improvin g recovery of nerve function. METHODS:The experiment was conducted in the Center for Experimental Animals o f Guangdong Medical College from September 2003 to April 2004.Forty- eight SD r ats in either gender weighing from 240 to 260 g were randomly divided into three groups:gene modification group,OEC transplantation group and SCI group with 16 rats in each group.All the rats were made into the models of spinal cord.Rats in gene modification group were transplanted with NGF and BDNF genes modified OECs ,and those in OEC transplantation group with OECs alone.The rats in SCI group we re treated with nothing.At 12 weeks after transplantation,the protein expression of Bcl- 2(an apoptosis suppressor gene),bax and Fas(two apoptosis induction ge nes) were mainly observed by detecting the apoptosis of spinal sections from the injured spinal cord with immunohistochemical method and terminal deoxynucleotid yl transferase- mediated dUTP nick end labeling(TUNEL) so as to evaluate the ef fect of NGF and BDNF genes modified OECs or modified OECs in suppressing and ind ucing apoptosis. RESULTS:All the 48 rats were included in the result analysis.① The results s hown by TUNEL:The quantity of apoptotic cells in the gene modification group was lower than that in OEC transplanation group,and the highest in the SCI group.② The results shown by immunohistochemistry:The protein expression of Bcl- 2 was the highest in the gene modification group(20.79± 6.40),followed by OEC transp lantation group(13.54± 3.66),and the lowest in the SCI group(9.34± 2.12)(P< 0. 05);and the protein expression of Fas and bax was the highest in SCI group(24.98 ± 4.32,40.48± 2.05),followed by OEC transplantation group(18.92± 4.74,25.54± 3.82) and the lowest in gene modification group(11.78± 3.81,16.87± 3.30)(P C

关 键 词：神经生长因子 基因修饰 脊髓损伤 脑衍生 嗅鞘细胞移植 原位末端标记法 Bcl-2蛋白表达 嗅神经鞘细胞移植 细胞凋亡抑制基因 神经营养因子 免疫组织化学法 BAX蛋白表达 神经功能恢复 实验动物中心 凋亡诱导基因 神经细胞凋亡 

分 类 号：R651.3[医药卫生—外科学] R735.7[医药卫生—临床医学]