骨髓间充质干细胞移植治疗大鼠脊髓损伤的神经分化研究  

Effect of mesenchymal stem cells on spinal cord injury transplantation in a rat model

作　　者：莫翠萍 王家传[1] 周光前[1] 姚晓璐[2] 龚飞鹏[2] 陈钢[2] 

机构地区：[1]深圳大学医学部深圳抗衰老与再生医学重点实验室,518000 [2]江西省人民医院骨一科,南昌330000

出　　处：《中华细胞与干细胞杂志（电子版）》2015年第4期33-39,共7页Chinese Journal of Cell and Stem Cell（Electronic Edition）

摘　　要：目的探讨骨髓间充质干细胞(BMSC)对移植脊髓损伤(SCI)模型神经再生修复的作用及其机制。方法 (1)分离原代SD大鼠BMSC;(2)体外诱导BMSC向神经分化,应用免疫荧光技术检测神经诱导分化后的BMSC神经标志Nestin、Neu N的表达;(3)运用改良的Allen撞击装置制备SD大鼠SCI模型,成年雌性SD大鼠12只,随机分组:损伤对照(n=6),BMSC细胞移植组(n=6),并选择在SCI后半小时内在蛛网膜下腔原位移植1×106 BMSC细胞注射治疗,对照组原位注射10μl PBS作为对照。每周对SCI大鼠进行BBB运动功能行为学评价。(4)在治疗后1个月处死SCI大鼠取脊髓样本进行冰冻切片检测Nestin、NeuN神经标记物表达情况,从而评判BMSC对SCI的治疗效果。计量资料结果服从正态分布、方差齐性时,采用t检验;若不服正态分布,采用KruskalWallis H秩和检验。结果 (1)分离的BMSC纯度高、生物学特征稳定。(2)BMSC在体外神经诱导环境下可分化为神经细胞,对比正常对照组,神经诱导组Nestin与Neu N的表达具有统计学差异(t=11.49、6.76,P<0.05)。(3)BMSC移植治疗SCI大鼠运动行为学功能显著改善,移植组比对照组治疗5周后的BBB评分具有统计学意义(t=5.59,P<0.05);损伤导致组织形态学出现脊髓白质灰质结构性损毁,神经细胞大量丢失,而BMSC移植组Nestin、GFAP与Neu N表达细胞均较损伤组差异有统计学意义(t=4.74、6.59、15.46,均P<0.05)。结论 BMSC移植可促进SCI后神经细胞的存活与再生分化,在一定程度上促进SCI脊髓组织功能的恢复。Objective To investigate the effects of marrow mesenchymal stem cells （BMSC） on spinal cord injury （SCI） and the underlying mechanisms. Methods BMSC were induced for neural differentiation in vitro for 7 days and Nestin and NeuN were detected. Spinal cord injury model was established by Allen＇s impact device. 1 × 10^6 BMSC were administered by subarachnoid injection in 30 minutes after SCI, followed by perfusion at 1 month. Spinal cord tissues were obtained and frozen tissue sections were prepared for the histological and immunochemical examinations for the expression of neural marker molecules, including Nestin and NeuN.The results of measurement obey the normal distribution and homogeneity of variance, using t test. If refuse to obey normal distribution, use Kruskal Wallis H rank and inspection. Results BMSC could be obtained with stable biological properties. A significant differences of Nestin and NeuN expression in BMSC-induced cells were detected （P 〈 0.05）. In the vivo experiments, BMSC infusion significantly improved spinal cord motor function accompanied by higher expression of Nestin and NeuN levels in the BMSC group compared to the vehicle group （P 〈 0.01）.Conclusions BMSC transplantation improved the outcomes of SCI probably via regeneration of neural cells from grafted BMSC.

关 键 词：骨髓 间质干细胞 脊髓损伤 神经再生 

分 类 号：R651.2[医药卫生—外科学]