低剂量纳曲酮治疗实验性自身反应性脑脊髓炎的初步研究

摘要

目的 通过诱导小鼠实验性自身反应性脑脊髓炎(EAE)模型,观察应用低剂量纳曲酮(LDN)干预对EAE发病及炎症反应的影响,从而探究LDN是否对EAE小鼠具有保护作用.方法 选取30只C57小鼠,随机分为3组,即EAE+ LDN组、EAE+ PBS组及健康对照组,其中,EAE+ LDN组在诱导EAE模型后给予0.2mL的LDN (0.1 mg/kg)腹腔注射,EAE +PBS组在诱导EAE模型后给予等量的PBS腹腔注射,健康对照组为未诱导EAE组.观察各组动物发病情况,每日监测小鼠体重变化,对小鼠进行行为学评分,苏木精-伊红(HE)染色法观察各组小鼠脑组织炎性细胞浸润情况,酶联免疫(ELISA)法监测血清中促炎性细胞因子水平变化.结果 LDN能有效延迟EAE小鼠的发病时间,并显著缓解发病过程中的体重下降及神经功能障碍;HE染色可见LDN干预能有效减轻EAE小鼠脑组织中的炎症反应,并能降低血清中促炎性细胞因子IFN-γ、IL-12、IL-17水平.结论 LDN能显著缓解EAE模型的发病,并减轻EAE发病过程中的炎性反应.%Objective To observe the effect of low-dose naltrexone (LDN) on the pathogenesis and the inflammatory response of mice model with the experimental autoimmune encephalomyelitis (EAE),and explore whether LDN had a protective effect on EAE mice.Methods Thirty C57 mice were randomly divided into EAE + LDN group,EAE + PBS group and healthy control group.EAE + LDN group was injected with 0.2 mL LDN (0.1 mg/kg) intraperitoneally,EAE + PBS group received intraperitoneal injection of equal volume of PBS,and healthy control group was unimmunized.The onset time of the disease,body weight and behavioral score were observed in the three groups.Hematoxylin-eosin (HE) staining was used to observe the infiltration of inflammatory cells in the brain tissue of mice,and the level of pro-inflannatory cytokines in serum was monitored by enzyme-linked immunosorbent assay (ELISA).Resuits LDN could significantly delay the onset time of EAE mice,and alleviate the weight loss and the neurological dysfunction during the process of disease.The intervention of LDN could effectively reduce the inflammatory reaction in the brain tissue of EAE mice by HE staining and decrease the expression of IFN-γ,IL-12 and IL-17 in serum.Conclusion LDN can alleviate the symptoms of EAE model significantly,and reduce the inflammatory reaction in the process of EAE disease.