MSCs移植并GS对大鼠脊髓损伤后期修复的影响  

EFFECT OF MESENCHYMAL STEM CELL TRANSPLANTATION COMBINED WITH GROWTH FACTOR ADMINISTRATION ON LATE-PHASE RECOVERY OF RATS WITH SPINAL CORD INJURY

作　　者：倪博文[1] 邹云雯[1] 王志杰[1] 褚言琛[2] 姚如永[3] 

机构地区：[1]青岛大学医学院附属医院脊柱外科,山东青岛266003 [2]青岛大学医学院附属医院创伤外科,山东青岛266003 [3]青岛大学医学院附属医院中心实验室,山东青岛266003

出　　处：《齐鲁医学杂志》2011年第4期319-321,324,共4页Medical Journal of Qilu

基　　金：国家自然科学基金资助项目(81071482)

摘　　要：目的观察骨髓间充质干细胞（MSCs）移植联合应用促细胞生长物质（GS）对脊髓损伤后期大鼠运动功能修复的影响。方法应用全骨髓法分离培养MSCs,10 mg/L的BrdU标记细胞核。将成年雄性Wistar大鼠36只,以改良Allen打击法制备T10脊髓损伤模型,制模后2周随机分为MSCs＋GS组、MSCs组与对照组,每组12只,伤后2周各组损伤处分别注入MSCs＋GS、单纯MSCs、培养液。分别于伤后1、2、3、4、5、6周进行BBB评分;损伤后6周处死大鼠,取损伤段脊髓及其上下各1 cm组织,行苏木精-伊红染色及SABC免疫组织化学方法染色,观察损伤脊髓组织病理变化和BrdU阳性细胞及GAP-43的相对表达。结果制模后1~2周3组大鼠后肢运动功能BBB评分未见明显差异（F=0.322、0.044,P〉0.05）,3~6周MSCs＋GS组大鼠后肢运动功能BBB评分较MSCs组和对照组高（F=13.729~97.187,P〈0.05）;MSCs＋GS组大鼠脊髓损伤中心及头、尾端均可见BrdU染色阳性细胞。同时,MSCs＋GS组及MSCs组损伤节段脊髓内GAP-43的表达面积明显多于对照组,MSCs＋GS组多于MSCs组。结论 MSCs移植联合GS促进大鼠损伤脊髓结构和功能恢复的效果明显优于单纯细胞移植,两者联用具有协同效应。Objective To investigate the effect of mesenchymal stem cell（MSCs） transplantation combined with growth factor（GS） administration on the recovery of rat motor function at late stage of spinal cord injury. Methods MSCs were isolated and cultured with cell attachment method.BrdU（10 mg/L） was used for labeling the nucleus of MSCs.Modified Allen＇s method was used to create a model of T10 spinal cord injury in 36 adult male Wistar rats.Two weeks later,the models were equally randomized to MSCs＋GS group,MSCs group and control group,and an injection of MSCs＋GS,MSCs and culture medium was given to the sites of injury,respectively.The rats were scored with Basso,Beattie and Bresnahan（BBB） locomotor rating scale 1,2,3,4,5 and 6 weeks after injury,and sacrificed after the last scoring.Tissue specimens from injured segment and from the area 1 cm above and below the injured region were processed for HE staining and SABC immunohistochemistry staining.Pathological changes in injured spinal cord,BrdU positive cells and GAP-43 relative expression were observed. Results One to two weeks after model establishment,there was no significant difference in BBB scores of rat hindlimb motor function between the three groups（F=0.322,0.044;P〉0.05）;three to six weeks after injury,BBB scores of rat hindlimb motor function in MSCs＋GS group showed greater improvement compared with MSCs group and control group（F=13.729-97.187,P〈0.05）.BrdU-positive cells were found at the center and both ends of the injured spinal cord in MSCs＋GS group.The expression of growth associated protein-43（GAP-43） in the injured spinal segment in both MSCs＋GS group and MSCs group were greater than that of the control group.Furthermore,GAP-43 expression was greater in MSC＋GS group than that in MSCs group. Conclusion The efficacy of combination therapy of MSCs transplantation and GS treatment is superior to MSCs transplantation alone in terms of promoting the structure and function of spinal cord.

关 键 词：脊髓损伤 间质干细胞移植 大鼠 

分 类 号：R349.7[医药卫生—基础医学]