新型Rho激酶抑制剂FSD-C11对EAE的免疫调节作用  

The mechanism of novel Fasudil derivative FSD-C11 in the treatment of experimental autoimmune encephalomyelitis

作　　者：于婧文[1] 李艳花[1] 张海飞[2] 尉杰忠[1] 刘春云[1] 肖保国[1,3] 马存根[1,4] 

机构地区：[1]山西大同大学脑科学研究所,037009 [2]山西省大同市第五人民医院神经科,037009 [3]复旦大学华山医院神经病学研究所,上海200025 [4]山西中医学院"2011"协同创新中心/神经生物学研究中心,太原030619

出　　处：《免疫学杂志》2016年第8期651-655,共5页Immunological Journal

基　　金：国家自然科学基金(81272163);山西省国际科技合作项目(2013081058);山西省回国留学人员重点科研资助项目(2014-重点7);山西中医学院“2011”培育计划(2011PY-1);大同大学校科研项目(2015Q15)

摘　　要：目的探讨Rho激酶抑制剂Fasudil衍生物FSD-C11治疗实验性自身免疫性脑脊髓炎(EAE)的免疫调节机制。方法采用MOG35-55多肽建立C57BL/6小鼠EAE模型,随机分为FSD-C11组和Saline组,于免疫后第3天起腹腔注射FSD-C11化合物和Saline。免疫后28 d处死小鼠,FACS法检测脾组织CD4+T细胞亚群,ELISA法检测外周免疫系统中细胞因子的分泌情况,Western blot法测定脊髓ROCKⅡ、i NOS、Arg-1、TLR-2和TLR-4的蛋白表达。结果 FSD-C11干预EAE能够抑制脊髓中ROCKⅡ表达,减少外周CD4+IFN-γ+T细胞,增加CD4+IL-10+和CD4+CD25+T细胞(P<0.05),减少外周免疫系统炎性细胞因子IFN-γ、IL-17、IL-6、IL-1β和TNF-α的含量(P<0.05),而增加IL-10的含量(P<0.05),抑制脊髓组织中巨噬细胞标志蛋白i NOS表达、增加Arg-1的表达(P<0.05)。抑制脊髓组织中TLR-2和TLR-4蛋白表达(P<0.05)。结论 FSD-C11可调节外周免疫细胞活化和增殖,抑制外周免疫系统分泌炎性因子,增加保护性的细胞因子,改善炎性微环境,促进M1型巨噬细胞向M2型转化,控制CNS的炎性细胞侵润,从而达到减轻或改善EAE的临床症状。This study aimed to investigate the mechanisms of novel Fasudil derivative FSD-Cll in the treatment of experimental autoimmune encephalomyelitis （EAE）. Chronic EAE model was induced by MOG35-55 peptides in female C57BL/6 mice. FSD-Cll or physiological saline were injected intraperitoneally from day 3 post-immunization to day 27 post-immunization. On day 28 post-immunization, mice were sacrificed and spinal cords were obtained for Western blot. Splenocytes were separated, and CD4＋T cell subset were detected by flow cytometry. The levels of cytokine were analyzed by ELISA. Data showed that the administration of FSD-Cll inhibited the expression of ROCK II in spinal cords, decreased CD4＋IFN-γ＋ T cells, increased CD4＋IL-10＋T cells and CD4＋CD25＋T cells in the peripheral immune system, and inhibited inflammatory cytokines IFN-γ IL-17, IL-6, IL-Iβ and TNF-α, but enhanced IL-10 level. Furthermore, FSD-Cll also inhibited the expression of macrophages phenotypic protein iNOS, up-regulated Arg-1 expression, inhibited the expression of TLR-2 and TLR-4 in spinal cords. Taken together, the therapeutic effect of FSD-Cll on EAE may relate to adjustment of peripheral immune cell activation and proliferation, such as inhibiting inflammatory cytokine and enhancing anti-inflammatory cytokine, improving inflammatory microenvironment, shifting M1 to M2 macrophages, preventing inflammatory cells migration into CNS.

关 键 词：Fasudil衍生物FSD—C11 EAE 免疫调节 

分 类 号：R744.5[医药卫生—神经病学与精神病学]