不同途径联合移植骨髓间充质干细胞-硫酸软骨素酶治疗家兔脊髓损伤的疗效比较  

Co-transplantation of bone mesenchymal stem cells with chondroitin sulfate enzymes through different routes in treating spinal injuries in rabbits

作　　者：王晶[1] 蔡颖谦[2] 周晓忠[1] 王业杨[1] 李超[1] 徐汪洋[1] 孙鸿涛[1] 李高祥[1] 姜晓丹[2] 李贵涛[1] 

机构地区：[1]南方医科大学附属广东省第二人民医院骨科,广州510317 [2]南方医科大学珠江医院神经外科,广州510280

出　　处：《中华神经医学杂志》2014年第3期217-223,共7页Chinese Journal of Neuromedicine

基　　金：广东省科技计划项目（2011A032100001,20128031800266）;广东省医学科研基金（A2012138）

摘　　要：目的比较不同途径联合移植骨髓间充质干细胞一硫酸软骨素酶修复家兔脊髓损伤（sci）的疗效差异。方法24只SCI家兔按照随机数字表法分为损伤对照（IC）ffi、椎管内移植（IS）组、静脉移植（Ⅳ）组和动脉移植（认）组，每组6只。后3组分别于SCI后1周经相应途径联合移植骨髓间充质干细胞一硫酸软骨素酶，IC组仅制作损伤模型，不行其他处理。SCI前及SCI后1、2、3、4周通过动物行为学评估、神经电生理检测、荧光示踪技术及免疫组化等方法，比较4组家兔疗效差异。结果（1）改良Tarlov评分：SCI后3周，IA组Tarlov评分与IC组比较明显升高，差异有统计学意义（P〈0．05）；术后4周，各移植组Tarlov评分明显优于IC组，差异均有统计学意义（P〈0．05）。（2）HE染色：各组损伤处均有大量空泡形成，相对于IC组，各移植组损伤处空泡较小，并可见部分神经纤维束通过。（3）免疫组化：各移植组神经丝蛋白表达量均明显强于IC组；IS组及IA组表达量明显高于Ⅳ组，差异均有统计学意义（P〈0．05）。胶质纤维酸性蛋白阳性表达面积方面，各移植组均低于IC组；而与IS和队组相比Ⅳ组表达面积进一步减少，差异均有统计学意义（P〈0．05）。（4）荧光双染比较：SCI后4周，各移植组镜下均可见较多的PKH．DAPI阳性细胞。其中IS和认组定植率（28．50％、27．75％）相比Ⅳ组（21．75％）明显增高，差异有统计学意义（P〈0．05）；（5）皮层感觉诱发电位潜伏期及波幅比较：SCI后第3、4周，各移植组潜伏期及波幅均优于IC组。差异有统计学意义（P〈0．05）；其中SCI后4周，IS组及认组潜伏期及波幅更优于Ⅳ组，差异有统计学意义（P〈0．05）。结论3种途径移植治疗对于SCI均有明显疗效，其中肋间动脉移植途径及椎管内移植疗效更为明显。[Abstract] Objective To compare the effects of different treatment routes on spinal cord injuries （SCI） in rabbits by co-transplantation of bone mesenchymal stem cells （BMSCs） with chondroitin sulfate enzymes （ChABC）. Methods Twenty-four SCI rabbits were randomized into injury control group （IC）, intraspinal treatment group （IS）, intravenous treatment group （IV） and intraarterial treatment group （IA, n=6）; BMSCs combined with ChABC were transplanted to the spinal cord tissue through the above three routes one week after SCI, respectively. Animal behavior evaluation, neural electrophysiology examination, and fluorescence tracer and immunohistochemical methods were used to compare with curative effects of co-transplantation via different routes in rabbit spinal cord injuries before SCI, and 1, 2, 3 and 4 weeks after SCI. Results Modified Tarlov scale scores： three weeks after SCI, Tarlov scale scores in IA group significantly increased as compared with that of IC group （P〈0.05）; four weeks after SCI, three treatment groups recovered more significantly than IC group （P〈0.05）. Hematoxylin-Eosin （HE） staining showed that large vacuoles were found at the spinal cord tissues of the IC group, while smaller vacuoles and visible part of the nerve fibers passing through were noted at the three treatment groups. Immunohistochemical method indicated that the expression of neurofilament 200 （NF200） was obviously more intense and that of glial fibrillary acidic portein （GFAP） was significantly weaker in the three treatment groups as compared with those in IC group （P〈0.05）; IS group and IA group had more intense NF200 expression than that in IV group （P〈0.05）; for GFAP expression, the highest expression was found in IC group, while that expression in IS and IA group was weaker and group 1V was much obvious, with significant differences （P〈0.05）. Double staining comparison： four weeks after SCI, many PKH-DAPI positive cells were seen in the three treatment

关 键 词：脊髓损伤 骨髓间充质干细胞 硫酸软骨素酶 移植途径 

分 类 号：R651.2[医药卫生—外科学]