马索罗酚对实验性自身免疫性脑脊髓炎小鼠Th1/Th2细胞炎症因子平衡的影响

目的探讨马索罗酚对实验性自身免疫性脑脊髓炎(EAE)小鼠白细胞介素-4(IL-4)、IL-12、干扰素-γ(IFN-γ)表达的调节作用。方法将8~10周雌性C57BL/6小鼠54只随机分成对照组、模型组、治疗组。每组再随机均分为发病后10d及20d亚组,每亚组9只。采用皮下注射髓鞘少突胶质细胞糖蛋白35-55(MOG35-55)多肽0.1mL诱导EAE模型。自发病当天起,治疗组小鼠给予马索罗酚10mg/(kg·d)治疗,模型组及对照组给予等量5%二甲基亚砜(DMSO)10mL/(kg·d)处理。比较3组小鼠临床症状评分。应用实时定量PCR检测小鼠脊髓和脾组织中IL-4、IL-12、IFN-γmRNA表达水平。应用ELISA检测脑组织中IL-4、IL-12、IFN-γ蛋白表达水平。结果与模型组比较,治疗组小鼠临床症状较减轻(P0.05)。与模型组相比,治疗组小鼠10d时脊髓和脾组织IL-12、IFN-γmRNA表达水平降低(P0.05),IL-4mRNA水平增高(P0.05),脑组织IL-12、IFN-γ蛋白水平降低(P0.05),IL-4蛋白水平增高(P0.05);与模型组相比,治疗组20d时脊髓组织IL-12、IFN-γmRNA表达水平降低(P0.05),脑组织IFN-γ含量降低(P0.05)。结论马索罗酚可能通过降低脑、脊髓及脾组织中IL-12、IFN-γ表达,增加IL-4表达,调节Th1/Th2细胞炎症因子平衡,进而改善EAE小鼠疾病严重程度。

Effects of nordihydroguaiaretic acid on Th1/Th2 cytokine balance in experimental autoimmune encephalomyelitis mice

Objective To explore the regulation effect of nordihydroguaiaretic acid (NDGA ) on interleukin‐4 (IL‐4) ,interleukin‐12 (IL‐12) and interferon‐gamma (IFN‐γ) levels in experimental autoimmune encephalomyelitis (EAE) mice .Methods Fifty‐four female C57BL/6 mice were randomly divided into 3 groups:the control ,EAE and treatment groups .Each group was divided into 10 day and 20 day subgroups according to the onset time .There were 9 mice in each subgroup .In the EAE and treatment group ,the EAE models were induced with 0.1 mL myelin oligodendrocyte glycoprotein35‐55 (MOG35‐55) (5 mg/mL) by subcutaneous injection .Normal saline was used in the control group . The clinical scores of the mice were recorded and analyzed .For treatment ,daily injection of NDGA at dose of 10 mg/ (kg ? d) was initiated on the day when clinical scores of mice≥1 .The EAE group and the control groups were given 5% dimethyl sulphoxide (DMSO) at the dose of 10 mL/ (kg?d) .Clinical symptom scores of the three groups were compared .IL‐4 ,IL‐12 ,IFN‐γmRNA levels in the spinal cord and spleen were detected by qRT‐PCR ,and IL‐4 ,IL‐12 ,IFN‐γ protein expression in the brain was detected by ELISA .Results Compared with the EAE group ,the clinical score in the treatment group declined (P< 0.05) .Compared with the EAE group ,the expression levels of IL‐12 ,IFN‐γmRNA decreased in the spinal cord and spleen and IL‐4 mRNA increased in the treatment group (P<0.05) ,the expression of IL‐12 and IFN‐γ in the brain decreased and IL‐4 increased in the treatment group at 10 d (P<0.05) .Compared with the EAE group ,the expression levels of IL‐12 ,IFN‐γ in spinal cord decreased in the treatment group ( P< 0.05 ) , and the expression level of IFN‐γ in brain decreased at 20 d ( P< 0.05 ) . Conclusions NDGA attenuates disease severity by down‐regulating the expression levels of IL‐12 and IFN‐γand up‐regulating IL‐4 in the spinal cord ,spleen and brain of EAE mice .