硫酸软骨素酶ABC联合骨髓间充质干细胞移植对脊髓损伤后神经功能的恢复作用  

Effect of bone mesenchymal stem cell transplantation combined with chondroitinase ABC on nerve function recovery in rats after spinal cord injury

作　　者：李敬花[1] 张涛[1] 张红军[1] 余雨[1] 卢旻鹏[1] 邵高海[1] 

机构地区：[1]重庆医科大学附属永川医院骨科,重庆402160

出　　处：《第三军医大学学报》2015年第2期122-127,共6页Journal of Third Military Medical University

基　　金：重庆市卫生局医学重点研究项目(2012-1-060)~~

摘　　要：目的探讨骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)移植联合硫酸软骨素酶ABC(chondro-itinrase ABC,Ch ABC)降解对脊髓损伤后神经功能修复作用。方法成年雄性SD大鼠60只,制作脊髓损伤模型并按随机数字表法分成4组(n=15):假手术组(A组)、Ch ABC降解组(B组)、BMSCs移植组(C组)、Ch ABC联合BMSCs移植组(D组)。4组分别在术后7、14 d观察记录动物情况并处死动物进行标本检测。伤后7、14 d,BBB评分评价大鼠后肢运动感觉功能;伤后7 d TUNEL法检测神经细胞凋亡情况;伤后14 d,HE染色测损伤区总面积;免疫组化法观察胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)、神经丝蛋白-200(NF-200)和勿动蛋白A(Nogo-A)的表达。结果伤后7 d各组BBB评分比较差异无统计学意义(P>0.05);伤后14 d,4组BBB评分分别为(6.33±1.00)、(7.25±0.96)、(7.33±0.58)、(9.75±0.96)分。D组BBB评分明显高于其他3组(P<0.05);伤后14 d,HE染色结果显示,4组损伤区均形成空洞,4组空洞相对面积分别为(4.44±0.18)、(3.70±0.21)、(3.23±0.22)、(2.33±0.31)mm2,D组空洞面积显著小于其他3组(P<0.05);伤后14 d GFAP/Nogo-A免疫组化染色结果显示D组GFAP/Nogo-A表达低于他3组(P<0.05);NF-200免疫组化染色结果显示D组阳性表达显著高于其他3组(P<0.05);伤后7 d TUNEL法检测结果显示D组阳性细胞数显著低于其他3组(P<0.05)。结论 Ch ABC联合BMSCs移植可以溶解胶质瘢痕,减少神经元细胞凋亡,促进神经纤维再生并越过脊髓损伤区域,对脊髓损伤后有明显的修复作用。Objective To explore the effect of bone mesenchymal stem cells （BMSCs） transplantation combined with chondro-itinrase ABC （ChABC） on the recovery of nerve function after spinal cord injury in rats. Methods A total of 60 adult male SD rats were selected, and the spinal cord injury model was established. The rats were randomly divided into 4 groups （ 15 rats per group） ＋ a control group （group A）, a BMSCs transplantation group （group B）, a ChABC injection group （group C）, and a ChABC ＋ BMSCs transplantation group （group D）. The rat conditions on the 7th and 14th days after injury were recorded, and the rats were executed to detect specimens. Basso-beattie-bresnahan （BBB） score was used to evaluate the rat hind leg movement functions. On the 7tb day after injury, the level of neuron apoptosis was detected with TUNEL assay, and on the 14th day after injury, HE staining was used to calculate the total area of the injured zones. Immunohistochemical assay was employed to examine the expression of glial fibrillary acidic protein （GFAP）, neurofilament-200 （NF-200） and Nogo-A protein in the injured spinal cord. Results On the 7th day after injury, no significant differences in the BBB scores wre found among the 4 groups （P= 0. 892）. On the 14th day after injury, the BBB scores of groups A to D were 6.33 ＋ 1.00, 7.25 ＋0. 96, 7.33 ＋ 0.58, and 9.75 _＋ 0.96, respectively, and the BBB score of group D was obviously higher than that of the other three groups （P 〈 0. 05 ）. On the 14th day after injury, HE staining showed that the cavities were formed in the injured zones, and the cavity areas of groups A to D were： 4.44 ±0.18, 3.70 ±0.21, 3.23 ± 0.22, and 2.33 ±0.31, respectively. The injured area of group D was significantly smaller than that of the other three groups （P 〈 0. 05 ）. On the 14th day after injury, the immunohistochemical assay showed that the expression of GFAP/Nogo-A in group D was obviously lower than that of the other three groups （ P �

关 键 词：脊髓损伤 ChABC BMSCS GFAP NF-200 细胞凋亡 

分 类 号：R345[医药卫生—基础医学] R617