人羊膜间充质干细胞对大鼠实验性自身免疫性脑脊髓炎的疗效及免疫调节作用  

Therapeutic effects and immunological regulation of human amniotic mesenchymal stem cells on rats with experimental autoimmune encephalomyelitis

作　　者：方宁[1] 李丹[1] 余丽梅[1] 章涛[1] 赵春华[2] 陈代雄[1] 

机构地区：[1]遵义医学院附属医院贵州省细胞工程重点实验室,563003 [2]中国医学科学院基础医学研究所组织工程中心,北京100005

出　　处：《免疫学杂志》2014年第1期6-12,共7页Immunological Journal

基　　金：国家科技重大专项(2009ZX09503-025);贵州省科技计划(黔科合计字2051)

摘　　要：目的探讨人羊膜间充质干细胞(hAMSCs)对大鼠实验性自身免疫性脑脊髓炎(EAE)模型的治疗效应和免疫调节作用。方法分离、培养hAMSCs,流式细胞术(FCM)检测其免疫表型;用豚鼠脊髓匀浆+完全弗氏佐剂+百日咳毒素复制大鼠EAE模型;模型动物随机分为正常对照组、模型组、培养基组、hAMSCs治疗组和泼尼松治疗组,每组6只。hAMSCs治疗组经双侧侧脑室移植5×105个hAMSCs;采用Kono评分法动态观察行为学变化;采用HE染色、免疫荧光染色观察脑和脊髓病理改变以及hAMSCs在脑和脊髓内的分布;采用FCM检测外周血淋巴细胞亚群,采用流式微球阵列法(CBA)检测血浆细胞因子的含量。结果 hAMSCs治疗后EAE大鼠行为学评分逐渐减低(P<0.01),脑和脊髓炎性细胞浸润减轻。hAMSCs移植第3周,大鼠脑和脊髓组织可见较多人细胞核抗原阳性细胞即hAMSCs存活;与模型组和培养基组相比,hAMSCs治疗组外周血Treg、Th2、Tc1、Tc2淋巴细胞百分比明显升高(P<0.01或P<0.05),而Th17细胞显著减少(P<0.01);血浆Th2型细胞因子IL-4和IL-10含量升高(P<0.05或P<0.01),而Th1型细胞因子IL-2和IFN-γ含量降低(P<0.05或P<0.01)。结论 hAMSCs能改善大鼠EAE的神经功能,减轻神经组织的免疫损伤,其机制可能主要与上调FoxP3+Treg细胞和下调Th17细胞有关。This study aimed to explore the therapeutic effects and immunological regulation of human amniotic mesenehymal stem cells （hAMSCs） on rats with experimental autoimmune encephalomyelitis （EAE）. Firstly, the hAMSCs were isolated from human amnion and cultured in vitro, and their phenotype was analyzed by flow cytometry （FCM）, while EAE was induced in Wistar rats with guinea pigs-spinal cord homogenate, complete Freund＇s adjuvant, and Bordetella pertussis toxin. Then rats were randomly divided into 5 groups： normal controls, untreated group, hAMSCs treated group, medium treated group, and prednisone treated group. In hAMSCs treated group, 5 xl05 of hAMSCs was injected into the lateral cerebroventricle of the rats with EAE. Therapeutic effects of hAMSCs on EAE were assessed by observing the behavior changes and the pathological changes in the brain and spinal cord of the rats, while the distributions of hAMSCs in injury brain and spinal cord were detected by immunofluorescence staining. The lymphocyte subsets in peripheral blood mononuclear cells were determined by flow cytometry; and the concentrations of plasma cytokines were determined by cytometric bead array. Results showed that the behavior scores of hAMSCs treated rats were reduced gradually （P 〈 0.01）. At week 3 post treatment, hAMSCs treated animals showed an improvement in inflammatory reaction in the brain and spinal cord, and the percentages of Treg, Th2, Tc｜ and Tc2 lymphocyte subsets were increased obviously （P 〈 0.05 or P 〈 0.0!）, while Thl7 cells were decreased significantly （P〈0.01）; furthermore, the concentrations of IFN--~ and IL-2 in the plasma were decreased （P〈 0.05or P〈0.01）, but IL-4 and IL-10 increased （P〈0.05 or P〈O.01）, as compared with other group. can improve neural function of rats with EAE and alleviate immunopathologic damage of mechanism might relate to Foxp3~ Treg up-regulation, and Th 17 cells down-regulation. In conclusion, hAMSCs neural tissues, and the

关 键 词：人羊膜间充质干细胞 实验性自身免疫性脑脊髓炎 免疫调节 大鼠 

分 类 号：R744[医药卫生—神经病学与精神病学]