盐酸法舒地尔抑制小鼠EAE作用及机制研究

Effect and Mechanism of Fasudil Hydrochloride on Experimental Autoimmune Encephalomyelitis Mice

目的探讨盐酸法舒地尔(Fasudil hydrochloride)对实验性自身免疫性脑脊髓炎(EAE)的抑制作用和可能的机制.方法按平均体重将C57BL/6雌性小鼠分为EAE组、盐酸法舒地尔治疗组和佐剂组,用髓鞘少突胶质细胞糖蛋白35-55多肽(MOG35-55)诱导EAE模型,佐剂组以等量生理盐水代替.治疗组于免疫后6d开始按50mg(/kg·d)腹腔注射盐酸法舒地尔,EAE组和佐剂组用等量的生理盐水作为对照.比较各组小鼠的各种表现,并进行症状评分.免疫后18～20d取各组小鼠,制备脾脏淋巴细胞悬液,用MTT法检测淋巴细胞的增殖情况,用ELISA法检测并比较各组淋巴细胞培养上清液中细胞因子INF-γ、IL-17、IL-10的水平.结果盐酸法舒地尔能够显著改善EAE的症状,减轻中枢神经系统(CNS)炎性浸润及髓鞘脱失等病理变化,并且可抑制脾脏淋巴细胞分泌INF-γ、IL-17(P<0.05),促进IL-10的分泌(P<0.05).结论盐酸法舒地尔可以有效抑制小鼠EAE,这种抑制可能与其对脾淋巴细胞中INF-γ、IL-17表达的下调和IL-10表达的上调作用有关.

Objective To investigate the suppressive effect and the possible mechanism of Fasudil hydrochloride on Experimental autoimmune encephalomyelitis （EAE） mice.Methods All female C57BL/6 mice were divided by average body weight into EAE group,Fasudil hydrochloride treated group and Adjuvant group.Mice in EAE and treated groups were immunized subcutaneously with myelin oligodendrocyte glycoprotein peptide （MOG35-55）.At the same time,adjuvant group was injected with normal saline instead of MOG35-55.Fasudil hydrochloride was intraperitoneally injected in the dosage of 50 mg（ kg·d）in treated group on day 6 post-immunization （p.i）,and mice in EAE and adjuvant group injected with normal saline as control.Manifestations of each groups were observed and compared.On day 18p.i,splenic lymphocytes were isolated under asepsis in each group respectively,and splenic lymphocyte suspensions were prepared.The proliferation assay was determined by MTT,and the supernatants were harvested for the detection of INF-γ,IL-17 and IL-10 by ELISA.Results Fasudil hydrochloride may significant ameliorate symptoms of EAE mice,lessen inflammatory cell infiltration and demyelination,suppress the secretion of INF-γ,IL-17 （P0.05）and promote the secretion of IL-10 （P 0.05）.Conclusions Fasudil hydrochloride can actively suppress EAE,which perhaps relates to down-regulation of INF-γ,IL-17 and up-regulation of IL-10.