两种干细胞对颅脑创伤炎症反应调控作用的对比研究  

Comparison on immunomodulatory effects of 2 kinds of transplanted stem cells on inflammation secondary to traumatic brain injury in mice

作　　者：高谋[1] 徐如祥[2] 杨志军[2] 董勤[3] 姚慧[2] 张洪钿[2] 杨阳[2] 刘宁[2] 陈晨[2] 朱建伟[2] 邹明明[1] 籍新潮[2] 

机构地区：[1]第三军医大学研究生管理大队,重庆400038 [2]北京军区总医院附属八一脑科医院神经外科,北京100700 [3]首都医科大学附属复兴医院神经内科,北京100038

出　　处：《第三军医大学学报》2015年第17期1697-1703,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81271316);军队"十二五"重大科技研究项目(BWS12J010)~~

摘　　要：目的比较神经干细胞（neural stem cells,NSCs）与间充质干细胞（mesenchymal stem cells,MSCs）移植对颅脑创伤（traumatic brain injury,TBI）后炎症反应的调控作用。方法将264只成年雄性C57BL/6小鼠按随机数字表法分为假手术组（54只）和TBI组（162只）,采用自由落体脑打击装置制备TBI模型并于伤后12 h进行NSS评分,将3~7分者按随机数字表法分为3组,每组54只,于伤后24 h将离体培养的同品系小鼠成体NSCs、MSCs和PBS分别定向注射到创伤模型鼠脑内。于移植后24、72 h和7 d处死动物,取脑组织病理切片染色,观察脑损伤修复和免疫细胞浸润情况;分别提取脑组织mRNA和蛋白检测细胞因子表达量。结果 1NSCs或MSCs移植7 d后,脑损伤较PBS处理组局限,组织内渗出减少。2NSCs或MSCs移植72 h和7 d后,脑组织内浸润的小胶质细胞/巨噬细胞数量明显少于PBS处理组（P〈0.05）;NSCs移植7 d后,脑内浸润的T细胞数量明显少于PBS处理组（P〈0.05）。3MSCs移植后IL-6和TNF-αmRNA和蛋白水平在各时间点均低于PBS处理组（P〈0.05）;NSCs移植后IL-10 mRNA水平在24 h和72 h明显高于PBS处理组（P〈0.05）,其蛋白水平在72 h和7 d也较PBS处理组和MSCs移植组明显升高（P〈0.05）。结论 NSCs与MSCs可影响TBI后炎症反应中免疫细胞浸润和细胞因子表达,二者调控作用既有相同之处,也有明显差异。Objective To compare the immunomodulatory effects of transplanted neural stem cells（ NSCs） and mesenchymal stem cells（ MSCs） on inflammation secondary to traumatic brain injury（ TBI）.Methods A total of 264 adult male C57 BL /6 mice were randomly assigned into sham operation group（ n =54） and TBI group（ n = 162）. Animal models of TBI were established using a free-falling weight drop device,and after 12 h,the mice with neurological severity score scores of 3- 7 points were selected for further research. The syngeneic adult NSCs,MSCs and PBS were injected into the brain of TBI mice with aid of a stereotaxic apparatus at 24 h after injury. Then,the mice were randomly sacrificed at 24 and 72 h and 7 d after injection,and the brain sections were stained for morphological analysis and evaluation of infiltrated immune cells. Besides,the expression of cytokines in the brain tissues were detected. Results In contrast to the PBS-treated group,the brain damage and tissue exudation were reduced obviously in the NSCs- or MSCstransplanted group after 7 d. The number of microglias / macrophages in the NSCs- or MSCs-transplanted group was significantly less than that in the PBS-treated group at 72 h and on 7 d（ P 0. 05）,and the number of T cells in the NSCs-transplanted group was significantly less than that in the PBS-treated group on the 7th day（ P 0. 05）. The mRNA and protein levels of interleukin（ IL）-6 and tumor necrosis factor（ TNF）-α in the MSCs-transplanted group were significantly lower than those in the PBS-treated group（ P 0. 05）. The IL-10 mRNA levels in the NSCs-transplanted group were significantly higher than those in the PBS-treated group at24 and 72 h（ P 0. 05）,and the IL-10 protein levels were also significantly higher than those in the PBStreated group and the MSCs-transplanted group at 72 h and on 7 d（ P 0. 05）. Conclusion Transplanted NSCs and MSCs can affect the infiltration of immune cells and the expression of cytokines so as to regulate the inflammation aft

关 键 词：神经干细胞 间充质干细胞 颅脑创伤 炎症反应 细胞因子 

分 类 号：R318.2[医药卫生—生物医学工程] R392.3[医药卫生—基础医学]