MSC-CNTF靶向性治疗变态反应性脑脊髓炎动物对病情和炎症的影响  

Bone marrow stromal cells transfected with ciliary neurotrophic factor gene ameliorates the symptoms and inflammation in C57BL/6 mice with experimental allergic encephalomyelitis

作　　者：陆正齐[1] 胡学强[1] 朱灿胜[1] 郑雪平[1] 王敦敬[2] 刘然义[3] 黄必军[3] 黄文林[3] 

机构地区：[1]中山大学附属第三医院神经科,广东广州510630 [2]徐州医学院附属第一医院神经科 [3]中山大学附属肿瘤医院实验研究部,广东广州510060

出　　处：《南方医科大学学报》2009年第12期2355-2361,共7页Journal of Southern Medical University

基　　金：广东省科技厅基金(2007B031506005)

摘　　要：目的探讨重组腺病毒转导的骨髓间充质细胞(MSC)导向的睫状神经营养因子(CNTF)基因靶向性治疗C57小鼠实验性变态反应性脑脊髓炎(EAE),对动物病情和和炎症的影响。方法用成功构建的Ad-CNTF-IRES-GFP载体,体外观察Ad-CNTF-IRES-GFP转染MSC(MSC-CNTF)对MSC表达CNTF的影响,再用MOG35-55建立C57BL/6小鼠的EAE模型,将MSC-CNTF移植治疗EAE小鼠,观察EAE动物病情评分;ELISA法观察外周血的促炎症因子TNF-α、IFN-γ,IL-12p35和抗炎症因子IL-10的水平,HE染色观察脑和脊髓炎症细胞浸润,用免疫荧光和免疫组化观察病变部位CD3(+)T细胞、炎性细胞因子TNF-α、IFN-γ的表达水平。结果(1)MSC-CNTF在MOI等于100的情况下,分泌的CNTF较DIL染色的MSC(MSC-DIL)、或Ad-eGFP转染的MSC(MSC-GFP)增高20倍(P<0.01)。(2)MSC-CNTF治疗的EAE小鼠病情显著减轻。不仅平均发病时间缩短,发病率下降,病情减轻,而且病情严重程度也明显减轻。(2)MSC-CNTF治疗明显减低EAE小鼠外周血TNF-α和IFN-γ水平,明显升高外周血IL-10水平。(3)MSC-CNTF治疗EAE小鼠可明显降低病变部位的炎症细胞数量和炎性细胞因子TNF-α水平,而对IFN-γ表达几乎影响不大。结论MSC-CNTF移植治疗的EAE动物病情明显减轻,其机制是:减低外周血TNF-α和IFN-γ水平,降低病变部位CD3(+)细胞数量和炎性细胞因子TNF-α水平。Objective To investigate the anti-inflammatory effect of bone marrow stromal cells （MSCs） transfected with recombinant adenovirus-mediated ciliary neurotrophic factor （CNTF） gene in C57BL/6 mice with experimental allergic encephalomyelitis （EAE）. Methods An adenovirus vector containing CNTF gene Ad-CNTF-IRES-GFP was constructed and transfected in the MSCs （MSC-CNTF）. After examination of CNTF expression, the transfected cells were transplanted in C57BL/6 mice with MOG 35-55-induced EAE, which were monitored for the changes in the symptoms scores. The levels of tumor necrosis factor-α （TNF-α）, inteferon-γ （IFN-γ）, interleukin-12P35 （IL-12P35）, and IL-10 in the peripheral blood of the mice were detected, and the number of MSC-CNTF cells in the spleen and spinal cord was counted. CD3＋ T cell infiltration and TNF-α and IFN-γ expressions in the lesions were also observed after the cell transplantation. Results CNTF gene transfection resulted in significantly increased CNTF expression in the MSCs. The mice receiving MSC-CNTF transplantation exhibited significantly improved symptoms with shortened disease course and lessened disease severity. The cell transplantation also resulted in significantly decreased peripheral blood TNF-α levels, ameliorated CD3＋T cell infiltrations and lowered TNF-α expression in the lesions, while the levels of IFN-γ underwent no significant changes. Conclusion Transplantation of CNTF gene-transfected MSCs results in decreased peripheral blood TNF-α and IFN-γ levels and reduced inflammatory cells, CD3-positive cells and TNF-α expression in the lesion of EAE, therefore providing better effect than MSCs in relieving the symptoms of EAE in mice.

关 键 词：睫状神经营养因子 重组腺病毒载体 多发性硬化 实验性变态反应性脑脊髓炎 骨髓间充质细胞 

分 类 号：R744[医药卫生—神经病学与精神病学]