法舒地尔治疗实验性自身免疫性脑脊髓炎小鼠与其调节T细胞免疫应答有关  

Treatment of experimental autoimmune encephalomyelitis mice by fasudil is related to regulation of T cell immune response

作　　者：刘春云[1] 郭尚德[1] 张年萍[1] 刘建春[2] 郭文娟[2] 于婧文[1] 肖保国[3] 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"2011"协同创新中心,山西太原030024 [3]复旦大学华山医院神经病学研究所,上海200040

出　　处：《中国新药与临床杂志》2016年第4期282-287,共6页Chinese Journal of New Drugs and Clinical Remedies

基　　金：国家自然科学基金面上项目(81272163);国家自然科学基金青年基金项目(81501032);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"培育计划项目(2011PY-1)

摘　　要：目的探讨法舒地尔对实验性自身免疫性脑脊髓炎(EAE)小鼠的治疗效果及作用机制。方法雌性C57BL/6小鼠,采用MOG_(35-55)多肽诱导建立慢性EAE模型,随机分为早期治疗组、晚期治疗组和EAE对照组。早期和晚期治疗组分别在免疫后第3日和疾病症状出现时,腹腔注射法舒地尔40 mg·kg^(-1)·d^(-1),观察各组小鼠体重变化和临床症状评分。免疫后第29日处死动物,取脊髓行HE染色和髓鞘染色;制备脾脏单个核细胞,应用流式细胞术检测T细胞亚群的变化;采用ELISA法观察细胞因子水平。结果法舒地尔治疗可推迟EAE发病时间(早期治疗,P<0.01),减轻临床症状(早期治疗,P<0.01;晚期治疗,P<0.05),减少炎性细胞浸润和髓鞘脱失(早期治疗,P<0.01;晚期治疗,P<0.05)。早期治疗组IL-17^+CD4^+T细胞比例显著低于EAE对照组(P<0.01),晚期治疗组IL-10^+CD4^+T细胞比例高于EAE对照组(P<0.01),同时早期和晚期治疗组IFN-γ^+CD4^+T细胞比例显著高于EAE对照组(均P<0.01)。此外,法舒地尔还可抑制MOG_(35-55)诱导的IL-17的分泌,促进IL-10的分泌。结论法舒地尔显示了治疗EAE的潜能,其机制可能与抑制Th17细胞分泌、促进Th2细胞表达有关。AIM To explore the therapeutic effect of fasudil and its possible mechanisms in experimental autoimmune encephalomyelitis（EAE） mice. METHODS Female C57BL/6 mice were immunized with MOG_（35-55） peptides to establish chronic EAE model. These mice were randomly divided into early treatment group in which fasudil 40 mg·kg^（-1）·d^（-1） was injected intraperitoneally on day 3 post- immunization, late treatment group in which fasudil was injected in a similar manner at onset of symptoms and EAE control group. Changes of body weight and clinical symptom scores were recorded in each group. On day 29 post-immunization, the mice were sacrificed and spinal cords were obtained for HE and LFB staining. The mononuclear cells（MNCs） from spleens were prepared and detected by flow cytometry to analyze T cells subgroups. ELISA method was used to examine cytokines production of MNCs. RESULTS The treatment of fasudil delayed onset of EAE（early treatment, P〈 0.01）, attenuated clinical symptoms（early treatment, P 〈0.01; late treatment, P〈 0.05） and reduced inflammatory cell infiltration and myelinoclasis（early treatment, P 〈0.01; late treatment, P 〈0.05）. The percentage of CD4~＋IL-17~＋T cells in early treatment group was significantly decreased compared with EAE control group（ P〈 0.01）. The percentage of CD4~＋IL-10~＋T cells in late treatment group was increased compared with EAE control group（P〈 0.01）. Meanwhile, the percentage of CD4~＋IFN-γ~＋T cells in early treatment and late treatment group was higher than that in EAE control group（ P〈 0.01）. Further studies showed that fasudil could inhibit the secretion of IL-17 and promote the production of IL-10 induced by MOG_（35-55）. CONCLUSION Fasudil shows the potential in treatment of EAE, possibly through inhibiting the secretion of Th17 cells and promoting the expression of Th2 cells.

关 键 词：法舒地尔 实验性自身免疫性脑脊髓炎 T细胞 免疫调节 

分 类 号：R744.5[医药卫生—神经病学与精神病学]