Fasudil联合骨髓源神经干细胞对EAE小鼠的神经保护作用  

Neuroprotective effect of fasudil combined with bone marrow-derived neural stem cells on mice with experimental autoimmune encephalomyelitis

作　　者：宋国娬 席国萍[1] 李艳花[1] 李加善[1] 刘建春[2] 柴智[2] 肖保国[3] 张光先 马存根[1,2] 

机构地区：[1]山西大同大学脑科学研究所,山西大同037009 [2]山西中医学院神经生物学研究中心,山西太原030024 [3]复旦大学华山医院神经病学研究所,上海200025 [4]托马斯·杰弗逊大学神经学系,美国宾夕法尼亚州费城19107

出　　处：《中国病理生理杂志》2017年第12期2113-2120,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81501198、No.81272163);山西大同大学青年科研基金资助项目(No.2013Q11);山西省回国留学人员重点科研资助项目(No.2014-重点7);山西中医学院“2011”培育计划项目(No.2011PY-1)

摘　　要：目的:探讨Rho激酶抑制剂法舒地尔(fasudil)联合骨髓源神经干细胞(bone marrow-derived neural stem cells,BM-NSCs)对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠的神经保护作用。方法:32只雌性C57BL/6小鼠(8~10周龄),用髓鞘少突胶质细胞糖蛋白35-55(MOG35-55)免疫,制备EAE,随机分为对照(dd H_2O)组、fasudil组、BM-NSCs组和fasudil+BM-NSCs组,并分别给予相应处理。免疫后检测小鼠临床症状和相关神经营养因子的表达,Image-Pro Plus 6.0软件进行阳性细胞计数,Graph Pad Prism 5软件统计分析。结果:与dd H_2O组比较,fasudil+BM-NSCs处理组明显延迟小鼠的平均起病时间,降低最高临床评分,并缓解EAE小鼠的临床症状;fasudil组、BM-NSCs组和fasudil+BM-NSCs组中脑源性神经营养因子、胶质细胞源性神经营养因子、神经生长因子、神经营养因子3和睫状神经营养因子阳性细胞数均有不同程度的增加,其中,fasudil+BM-NSCs组上述神经营养因子的表达明显多于dd H_2O组、fasudil组和BM-NSCs组(P<0.01)。结论:Fasudil联合BM-NSCs通过协同和叠加效应促进神经营养因子表达,改善中枢神经系统微环境,发挥神经保护作用,从而缓解EAE的临床症状。AIM ： To explore the neuroprotective effect of fasudil combined with bone marrow-derived neural stem cells （ BM-NSCs） on the mice with experimental autoimmune encephalomyelitis （EAE）. METHODS ： Female C57BL/6 mice （8-10 weeks old, n =32） were immunized with myelin oligodendrocyte glycoprotein 35-55 （MOG35-55 ） to establish chronic EAE model. The mice were randomly divided into control （ ddH20 ） group, fasudil group, BM-NSCs group, and fasudil ＋ BM-NSCs group. The clinical score and body weight were recorded every other day. The expression of neurotrophic factors was determined by immunofluorescence staining. RESULTS： In comparison with ddH20 group, fasud-il combined with BM-NSCs delayed onset and ameliorated severity of EAE. The numbers of brain-derived neurotrophic fac-tor, glial cell-derived neurotrophic factor, nerve growth factor, neurotrophin-3 and ciliary neurotrophic factor positive cells in fasudil group, BM-NSCs group and fasudil ＋ BM-NSCs group were all increased in various extents. In particularly, the expression of these neurotrophic factors in fasudil ＋ BM-NSCs group was significantly higher than that in the mice treated with fasudil or BM-NSCs alone （ P 〈 0. 01） . CONCLUSION ： Fasudil combined with BM-NSCs promotes the expression of neurotrophic factors and improves microenvironment of central nervous system, thus playing a positive role in neural restora-tion and regeneration through a synergistic and superimposed effect.

关 键 词：法舒地尔 骨髓源神经干细胞 实验性自身免疫性脑脊髓炎 神经营养因子 

分 类 号：R0[医药卫生] R329.5