间充质干细胞改善自身免疫性脑脊髓炎的作用研究  

The study on mesenchymal stem cells ameliorating experimental autoimmune encephalomyelitis

作　　者：张俊华[1] 郑配国[1] 

机构地区：[1]郑州大学第一附属医院检验科,450052

出　　处：《国际免疫学杂志》2015年第2期129-133,共5页International Journal of Immunology

摘　　要：目的探讨间充质干细胞（MSC）治疗自身免疫性脑脊髓炎（EAE）的机制。方法用MOG35-55肽和弗氏完全佐剂乳化剂诱导建立C57BL／6小鼠的EAE模型；分离纯化培养骨髓来源MSC细胞；临床评分和脊髓组织切片评估小鼠的发病情况；酶联免疫吸附试验（ELISA）检测EAE组，MSC治疗组和对照组小鼠外周血细胞因子TNF-α，IFN-γ，IL-4和TGF-β的含量；流式细胞术分析3组小鼠脾脏细胞中CD4^＋Foxp3^＋调节性T细胞（Treg）的比例变化。结果分离纯化C57BL／6小鼠MSC成功；MSC治疗组小鼠的临床评分明显降低，且脊髓组织切片显示T细胞浸润显著减少；外周血中细胞因子IL-4，TGF-β显著高于对照组（t=7．719、17．17，P均〈0．01）和EAE组（t=54．45、48．36，P均〈0．01），IFN-γ，TNF-α低于对照组（t=104．90、1．998，P均〈0．01）和EAE组（t=270．1、13．58，P均〈0．01）；脾脏细胞中CIM’Foxp3’Treg细胞的比例明显高于对照组（t=15．91，P〈0．01）和EAE组（t=33．39，P〈0．01）。结论MSC能够有效改善EAE小鼠的症状；其对EAE小鼠的治疗作用是通过上调抗炎细胞因子（IL-4，TGF-β）和Treg细胞并下调促炎因子（IFN-γ，TNF-α的水平，来发挥免疫调节作用。Objective To investigate the mechanism of mesenchymal stem cells （MSC） ameliorating experimental autoimmune encephalomyelitis （EAE）. Methods EAE model of C57BL/6 mice were established by the administration of MOG35-55/CFA emulsion. MSCs were purified from bone marrow of C57BL/6 mice and cultured. Then we recorded the clinical score and analyzed the pathological change of spinal cord to evaluate the state of EAE. The levels of TNF-α, IFN-γ, IL-4, and TGF-βcytokines in peripheral blood were evaluated by ELISA in EAE group, MSC-treated group, and the control group. CD4^＋ Foxp3 ^＋ Treg cells in these three groups were analyzed by flow cytometry. Results MSC were successfully purified and cultured. The clinical score and T-cell infiltration in spinal cord of MSC-treated mice reduced significantly. IL-4 and TGF-β in pe- ripheral blood of the MSC-treated mice were obviously higher than those in control group（ t = 7. 719,17. 17 ,P 〈 0. 01 ） and EAE group（ t = 54. 45,48.36, P 〈 0. 01 ）, while IFN-γ and TNF-α were significantly lower than those in control group （t = 104.90,1. 998, P 〈 0. 01 ） and EAE group （ t = 270. 10,13.58, P 〈 0.01 ）. The percent of CD4^＋ Foxp3^＋ Treg cells in spleen in the MSC-treated group was higher than that in control group（ t = 15.91 ,P 〈0. 01 ） and EAE group（ t = 33.39 ,P 〈0. 01 ）. Conclusion The transplantation MSC can effectively improve the condition of experimental autoimmune encephalomyelitis. MSC performs the function of immunoregulation on EAE through up-regulating the anti-inflammatory cytokines （IL4, TGF-β） and Treg cells and down-regulating the proinflammatory cytokines （ IFN-γ, TNF-α）.

关 键 词：间充质干细胞 实验性自身免疫性脑脊髓炎 CD4^+Foxp3^+Treg细胞 免疫调节 

分 类 号：R744.5[医药卫生—神经病学与精神病学]