BMSCs移植治疗大鼠脊髓损伤效果及局部细胞因子表达变化  

EFFECTS OF BONE MARROW MESENCHYMAL STEM CELLS TRANSPLANTATION FOR TREATING RAT SPINAL CORD INJURY AND CYTOKINE EXPRESSION AT INJURY SITES

作　　者：莫翠萍 任力杰[2] 赵振富[1] 周光前[1] 姚晓璐[3] 龚飞鹏[3] 陈钢[3] 

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,广东深圳518054 [2]深圳大学第一附属医院深圳市第二人民医院神经内科 [3]江西省人民医院骨一科

出　　处：《中国修复重建外科杂志》2016年第3期265-271,共7页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(81160156);江西省自然科学基金资助项目(20114BAB205091);深圳市重点实验室课题资助项目(zdsys20140509100746679);广东省自然科学基金资助项目(2013B021800102)~~

摘　　要：目的探讨BMSCs移植治疗大鼠脊髓损伤的效果及局部细胞因子的变化、作用。方法取SD大鼠采用全骨髓培养法分离培养BMSCs并传代,取第5代细胞采用腺病毒r Ad-EGFP进行转染标记。取12只SD大鼠,随机分为实验组及对照组(n=6)。两组大鼠采用改良Allen撞击装置制备T10脊髓损伤模型后,分别于脊髓损伤处注射10μL 1×106个标记的BMSCs(实验组)及PBS(对照组)。术后观察大鼠一般情况,于术后即刻及1、2、3、4、5周采用BBB评分法进行运动功能评分;术后5周处死大鼠取脊髓样本进行HE染色及神经巢蛋白(Nestin)、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)、神经元特异性核蛋白(neuron-specific nuclear protein,Neu N)免疫荧光染色观测,以及细胞因子抗体芯片检测。结果术后5周实验组及对照组各2只大鼠因尿路感染死亡,其余大鼠均存活至实验完成。除术后即刻两组BBB评分比较差异无统计学意义(P>0.05)外,1、2、3、4、5周实验组均显著高于对照组,比较差异有统计学意义(P<0.05)。术后5周,组织学观察示实验组细胞较对照组多且排列整齐;实验组Nestin、Neu N表达较对照组显著增加、GFAP表达较对照组显著降低,比较差异均有统计学意义(P<0.05)。细胞因子检测示,两组6个细胞因子表达存在差异;其中,实验组瘦素、睫状神经营养因子高于对照组,粒细胞和巨噬细胞集落刺激因子、TNF-α、IL-1β与基质金属蛋白酶组织抑制剂1低于对照组。结论 BMSCs移植可促进大鼠脊髓损伤后神经细胞的存活与再生,并调控细胞因子影响脊髓组织功能的恢复。Objective To investigate the effects of bone marrow mesenchymal stem cells（BMSCs） transplantation for treating spinal cord injury（SCI） in rat and the cytokine expression changes in the local injury tissues. Methods BMSCs were separated from Sprague Dawley（SD） rat and cultured with the whole bone marrow culture method. r Ad-EGFP was used to transfect the 5th generation BMSCs for green fluorescent protein（GFP） label. Twelve SD rats were randomly divided into experimental group（n=6） and control group（n=6）. After the T10 SCI model was established with Allen＇s impact device in 2 groups, 1×10^6 GFP-labeled BMSCs and PBS were administered by subarachnoid injection in situ in experimental group and control group, respectively. Basso-Beattie-Bresnahan（BBB） score was used to detect the motor function at immediat, 1, 2, 3, 4, and 5 weeks after SCI. At 5 weeks, the spinal cord tissues were harvested for the histological and immunofluorescent staining examinations to measure the expressions ofneural marker molecules, including Nestin, glial fibrillary acidic protein（GFAP）, and neuron-specific nuclear protein（Neu N）. Cytokine was analyzed with antibody array. Results At 5 weeks, 2 rats died of urinary tract infection in 2 groups respectively, the other rats survived to the end of experiment. BBB score of experimental group was significantly higher than that of control group at 1, 2, 3, 4, and 5 weeks（P〈0.05）. At 5 weeks, histological results showed that there were many cells with regular arrangement in the experimental group; there were less cells with irregular arrangement in the control group. Compared with the control group, Nestin and Neu N expressions significantly increased（P〈0.05）, and GFAP expression significantly decreased（P〈0.05） in the experimental group. Leptin and ciliary neurotrophic factor levels were higher in the experimental group than the control group, but granulocyte-macrophage colony-stimulating factor, tumor necrosis factor α, interleukin 1 β, an

关 键 词：脊髓损伤 BMSCS 细胞因子 神经再生 大鼠 

分 类 号：R651.2[医药卫生—外科学]