间充质干细胞联合白藜芦醇对实验性自身免疫性脑脊髓炎小鼠炎症的影响

摘要

目的 研究间充质干细胞联合白藜芦醇治疗缓解实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠模型疾病进程的机制,探讨联合治疗对EAE小鼠炎症的作用.方法 正常健康C57BL/6雌性小鼠经髓鞘少突胶质细胞糖蛋白35-55多肽(MOG35-55)诱导构建EAE小鼠模型,通过观察EAE小鼠的临床评分,判断EAE小鼠使用间充质干细胞联合白藜芦醇处理的效果.通过HE 染色和抗CD4的免疫组织化学检测单核细胞和T淋巴细胞对于脊髓组织的浸润.应用ELISA检测白藜芦醇和间充质干细胞处理后EAE小鼠血清中Th1型细胞因子γ干扰素(Interferon-γ,IFN-γ)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和Th2型细胞因子白细胞介素(interleukin,IL)-4、IL-10的表达量.结果 4组平均临床评分、最大临床评分、细胞浸润、CD4+T细胞浸润、IFN-γ、TNF-α mBM-MSCs处理组、白藜芦醇处理组、mBM-MSCs联合白藜芦醇处理组低于PBS处理组,mBM-MSCs联合白藜芦醇处理组低于mBM-MSCs处理组和白藜芦醇处理组,差异均有统计学意义(P<0.05);IL-4、IL-10 mBM-MSCs处理组、白藜芦醇处理组、mBM-MSCs联合白藜芦醇处理组高于PBS处理组,mBM-MSCs联合白藜芦醇处理组高于mBM-MSCs处理组和白藜芦醇处理组,差异均有统计学意义(P<0.05).结论 间充质干细胞联合白藜芦醇治疗可以显著延缓EAE疾病进程,同时联合治疗能够显著降低小鼠EAE模型的疾病严重性,从而改善EAE疾病的临床评分;可以降低EAE小鼠脊髓中的炎症细胞数目,并促进EAE小鼠Th1型向Th2型细胞因子的转变.%Objective To study in depth the machenism underlying the therapeutic efficiency of mesenchymal stem cell combined with resveratrol in encephalomyelitis(EAE), and to investigate the effect of mesenchymal stem cell combined with resveratrol on immune inflammation in EAE mice.Methods We investigate the therapeutic efficiency of bone marrow mesenchymal stem cell combined with resveratrol in experimental autoimmune encephalomyelitis by clinical score.The infliteration of immune cells such as monocytes and lymphocytes into brain was observed using HE staining and immunohistochemistry with anti-CD4 antibody.ELISA was conducted to detect the secretion of IFN-γ/TNF-α and IL-4/IL-10 inserum.Results The average clinical score, maximal clinical score, CD4+ T cell infiltration, IFN-γ, TNF-α were lower in mBM-MSCs group, resveratrol group, mBM-MSCs combined resveratrol group than that in PBS group.The mBM-MSCs combined resveratrol treatment group was lower than the mBM-MSCs treatment group and the resveratrol treatment group(P<0.05);The levels of IL-4 and IL-10 were higher in mBM-MSCs group, resveratrol group, mBM-MSCs combined resveratrol group than that in PBS group.Combination treatment group was higher than mBM-MSCs group or resveratrol group.The differences were statistically significant(P<0.05).Conclusion Combination treatment with mBM-MSCs and resveratrol can significantly delay the progression of EAE disease.Combined treatment can reduce the number of infla-mmatory cells in EAE mouse spinal cord and promote the transition of type Th1 to type Th2 cytokines in EAE mice.