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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。
Objective To investigate the therapeutic effect and mechanism of fasudil on experimental autoimmune encephalomyelitis (EAE) mice. Methods Female C57BL / 6 mice were induced by MOG_ (35-55) polypeptide to establish chronic EAE model and were randomly divided into early treatment group, advanced treatment group and EAE control group. In the early and late treatment groups, Fasudil 40 mg · kg -1 · d -1 was injected intraperitoneally on the 3rd day after the immunization and the symptoms of the disease were observed. Symptom score. Animals were sacrificed on the 29th day after immunization, and the spinal cord was taken for HE staining and myelin staining. Spleen mononuclear cells were prepared. The changes of T cell subsets were detected by flow cytometry. The levels of cytokines were measured by ELISA. Results Fasudil could delay the onset of EAE (early treatment, P <0.01), relieve clinical symptoms (early treatment, P <0.01; late treatment, P <0.05), decrease inflammatory cell infiltration and demyelination Early treatment, P <0.01; late treatment, P <0.05). The proportion of IL-17 ~ + CD4 ~ + T cells in early treatment group was significantly lower than that in EAE control group (P <0.01), and the proportion of IL-10 ~ + CD4 ~ + T cells in advanced treatment group was higher than that in EAE control group , While the proportion of IFN-γ ~ + CD4 ~ + T cells in early and late treatment groups was significantly higher than that in EAE control group (all P <0.01). In addition, Fasudil could also inhibit the secretion of IL-17 induced by MOG_ (35-55) and promote the secretion of IL-10. Conclusion Fasudil shows the potential of treating EAE. The mechanism may be related to inhibiting the secretion of Th17 cells and promoting the expression of Th2 cells.