| 摘要: |
| 目的 观察间充质干细胞(MSC)对实验性自身免疫性脑脊髓炎(EAE)的疗效并探讨其免疫调节机制.方法 用髓鞘少突胶质细胞糖蛋白(MOG35-55)和弗氏完全佐剂诱导建立C57BL/6小鼠EAE模型,随机分为MSC治疗组和EAE组.分离纯化培养GFP-C57BL/6 小鼠骨髓MSC细胞.在EAE诱导后的第15天,MSC治疗组以每只1×106/400 μL的量经尾静脉注射MSC细胞,EAE组尾静脉注射400 μL PBS.采用临床评分和脊髓组织切片评估小鼠的发病情况,ELISA检测小鼠外周血细胞因子肿瘤坏死因子α(TNF-α)、干扰素γ(IFN-γ)、白介素4(IL-4)和转化生长因子β(TGF-β)的含量,流式细胞术分析小鼠脾脏细胞中CD4+Foxp3 T细胞(Treg)的比例变化和GFP 细胞在受体小鼠体内的比例变化.结果 与EAE组相比,MSC治疗组小鼠的临床评分降低(P<0.05); 脊髓组织切片中T细胞浸润减少;血浆中细胞因子IL-4和TGF-β的含量升高,而IFN-γ和TNF-α降低;脾脏细胞中CD4 Foxp3 T细胞(Treg)的比例明显升高.移植10 d后MSC消失.结论 MSC移植对小鼠EAE疗效显著.MSC通过增加血浆中抗炎因子IL-4和TGF-β的含量,降低促炎因子IFN-γ和TNF-α的含量,从而促使原始T细胞向Treg细胞分化,发挥免疫调节作用, 且MSC消失后一段时间仍对EAE有一定治疗作用. |
| 关键词: 间质干细胞 实验性自身免疫性脑脊髓炎 调节性T淋巴细胞 免疫调节 |
| DOI：10.3724/SP.J.1008.2015.00034 |
| 投稿时间：2014-05-28修订日期：2014-10-30 |
| 基金项目:国家自然科学基金(81172835). |
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| Mesenchymal stem cells for treatment of experimental autoimmune encephalomyelitis and the immunoregulation mechanism |
| ZHANG Jun-hua,ZHENG Pei-guo,MA Xue-han,LIU Hong-chun*,MING Liang* |
(Department of Clinical Laboratory, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China
*Corresponding author) |
| Abstract: |
| Objective To observe the efficacy of mesenchymal stem cells (MSC) for treatment of experimental autoimmune encephalomyelitis (EAE) and the related immunoregulation mechanism. Methods EAE models of C57BL/6 mice were established with MOG35-55/CFA emulsion and the EAE mice were randomly divided into MSC treatment group and EAE model group. MSCs were purified and cultured from the bone marrow of GFP-C57BL/6 mice. On the 15th day of EAE model establishment, MSCs (1×106/400 μL) were injected via the tail vein for the MSC treated group, and 400 μL PBS were injected for the EAE model mice. Then we recorded the clinical scores and analyzed the pathological changes of spinal cord so as to evaluate the state of EAE in each group. The peripheral blood tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-4 (IL-4) and transforming growth factor-β (TGF-β) cytokines were evaluated by ELISA. The percent of CD4+Foxp3 Treg cells and GFP cells were analyzed by Flow cytometry. Results Compared with EAE model group, the MSC treated group had significantly decreased clinical score(P<0.05) and less T-cell infiltration in the spinal cord. MSC treated group had increased peripheral blood IL-4, TGF-β and decreased IFN-γ, TNF-α levels. Moreover, the MSC treated group also had greatly increased ratio of CD4 Foxp3 T cells (Treg). The MSC cells almost disappeared 10 days after transplantation. Conclusion Mesenchymal stem cell transplantation can effectively treat experimental autoimmune encephalomyelitis in mice. MSC may exert immunoregulation effect through increasing blood anti-inflammatory cytokines (IL-4, TGF-β) and decreasing proinflammatory cytokines(IFN-γ, TNF-α), which can prompt the naive cell differentiation into Treg cells. The effect of MSC remains even after disappearing for a certain time period. |
| Key words: mesenchymal stem cells experimental autoimmune encephalomyelitis regulatory T-lymphocytes immunoregulation |
