CAJ | 학술논문

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MOG 35-55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TN-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凶子TNF-α水平.
목적 탐토중조선병독전도적골수간충질세포(MSC)도향적첩상신경영양인자(CNTF)기인파향성치료C57소서실험성변태반응성뇌척수염(EAE),대동물병정화화염증적영향.방법 용성공구건적Ad-CNTF-IRES-GFP재체,체외관찰Ad-CNTF-IRES-GFP전염MSC(MSC-CNTF)대MSC표체CNTF적영향,재용MOG 35-55건립C57BL/6소서적EAE모형,장MSC-CNTF이식치료EAE소서,관찰EAE동물병정평분;ELISA법관찰외주혈적촉염증인자TNF-α、IFN-γ,IL-12p35화항염증인자IL-10적수평,HE염색관찰뇌화척수염증세포침윤,용면역형광화면역조화관찰병변부위CD3(+)T세포、염성세포인자TNF-α、IFN-γ적표체수평.결과 (1)MSC-CNTF재MOI등우100적정황하,분비적CNTF교DIL염색적MSC(MSC-DIL)、혹Ad-eGFP전염적MSC(MSC-GFP)증고20배(P<0.01).(2)MSC-CNTF치료적EAE소서병정현저감경.불부평균발병시간축단,발병솔하강,병정감경,이차병정엄중정도야명현감경.(2)MSC-CNTF치료명현감저EAE소서외주혈TN-α화IFN-γ수평,명현승고외주혈IL-10수평.(3)MSC-CNTF치료EAE소서가명현강저병변부위적염증세포수량화염성세포인자TNF-α수평,이대IFN-γ표체궤호영향불대.결론 MSC-CNTF이식치료적EAE동물병정명현감경,기궤제시:감저외주혈TNF-α화IFN-γ수평,강저병변부위CD3(+)세포수량화염성세포흉자TNF-α수평.
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.