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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1×106/400μL的量经尾静脉注射MSC细胞,EAE组尾静脉注射400μL 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10d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。
Objective To observe the therapeutic effect of mesenchymal stem cells (MSCs) on experimental autoimmune encephalomyelitis (EAE) and to explore its immunoregulatory mechanism. Methods C57BL / 6 mice model of EAE was induced by myelin oligodendrocyte glycoprotein (MOG35-55) and Freund’s complete adjuvant. The rats were randomly divided into MSC-treated group and EAE-treated group. Isolation and purification of GFP-C57BL / 6 mouse bone marrow MSC cells. On the 15th day after induction of EAE, MSC-treated mice were injected with MSC cells in the tail vein at an amount of 1 × 10 6/400 μL each, and 400 μL of PBS was injected into the tail vein of the EAE group. The clinical scores and spinal cord sections were used to assess the incidence of mice. ELISA was used to detect the levels of TNF-α, IFN-γ, IL-4 in the peripheral blood of mice, (TGF-β), the ratio of CD4 + Foxp3 + T cells (Treg) in spleen cells and the proportion of GFP + cells in the recipient mice were analyzed by flow cytometry. Results Compared with EAE group, the clinical scores of MSCs-treated mice decreased (P <0.05); the number of T cells infiltrated in spinal cord tissue decreased; the levels of IL-4 and TGF- γ and TNF-α decreased; the proportion of CD4 + Foxp3 + T cells (Treg) in spleen cells was significantly increased. MSC disappeared 10 days after transplantation. Conclusion MSC transplantation has a significant effect on EAE in mice. MSC can promote the differentiation of primary T cells into Treg cells by increasing the contents of anti-inflammatory cytokines IL-4 and TGF-β in plasma and decreasing the contents of proinflammatory cytokines IFN-γ and TNF-α After some time there is still some EAE therapeutic effect.