马索罗酚对实验性自身免疫性脑脊髓炎小鼠IL-6、IL-17、TGF-b表达的影响

目的观察马索罗酚(nordihydroguaiaretic acid,NDGA)对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠IL-6、IL-17、TGF-β表达的影响。方法采用MOG35-55多肽为抗原免疫C57BL/6小鼠制备EAE模型。小鼠随机分成对照组、模型组和治疗组,各组18只。治疗组给予马索罗酚10mg/(kg·d)干预,对照组及模型组给予等量溶剂干预。分别在发病后10 d、20 d进行神经功能评分后取材,采用实时定量PCR方法检测小鼠腰髓和脾组织IL-6、IL-17、TGF-bm RNA表达水平,采用ELISA方法检测脑组织IL-6、IL-17、TGF-b蛋白表达水平。结果模型组与治疗组发病10 d、20 d时神经功能评分为(7.22±1.10)、(4.22±0.97),(2.89±1.05)、(1.33±0.71),差异有统计学意义(P0.05)。治疗组与模型组比较结果如下。脊髓组织:1IL-6水平发病10 d时(8.54±2.49)、(14.07±3.19),差异有统计学意义(P0.05);发病20 d时(4.26±1.44)、(4.90±1.24),差异无统计学意义(P0.05)。2IL-17水平发病10 d时(25.50±11.77)、(47.59±8.01);发病20 d时(10.02±2.99)、(20.14±4.57),差异均有统计学意义(P0.05)。3TGF-b水平发病10 d时(0.74±0.11)、(0.04±0.01);发病20 d时(1.49±0.30)、(0.59±0.23),差异均有统计学意义(P0.05)。脑组织(pg/m L):1 IL-6水平发病10 d时(44.58±6.94)、(57.46±7.32);发病20 d时(35.85±9.58)、(51.29±2.09),差异均有统计学意义(P0.05)。2IL-17水平发病10 d时(515.90±63.10)、(899.89±242.39);发病20 d时(437.71±137.35)、(730.32±139.94),差异均有统计学意义(P0.05)。3TGF-b水平发病10 d时(170.88±24.69)、(132.85±20.27);发病20 d时(229.59±32.10)、(172.58±16.94),差异均有统计学意义(P0.05)。脾组织:1 IL-6水平发病10 d时(6.51±1.29)、(19.76±5.85),差异有统计学意义(P0.05);发病20 d时(3.13±0.78)、(4.14±0.58),差异无统计学意义(P0.05)。2IL-17水平发病10 d时(2.08±1.19)、(7.50±1.31);发病20 d时(0.67±0.41)、(2.75±0.44),差异均有统计学意义(P0.05)。3TGF-b水平发病10 d时(0.61±0.16)、(0.08±0.02);发病20 d时(1.77±0.39)、(0.50±0.33),差异均有统计学意义(P0.05)。结论马索罗酚可以改善EAE小鼠病情,其作用机制可能与下调IL-6、IL-17表达,上调TGF-b表达有关。

NDGA ameliorates experimental autoimmune encephalomyelitis via the regulation of IL-6, IL-17 and TGF-b

Objective To investigate the effects of nordihydroguaiaretic acid (NDGA) on the expression of IL-6, IL-17 and TGF-βin mice with experimental autoimmune encephalomyelitis (EAE). Methods C57BL/6 mice were immu? nized with MOG35-55 to induce EAE. The 54 mice were randomly and equally divided into control group, model group and treatment group. Mice in treatment group were daily administered with NDGA (10 mg/kg) by intraperitoneal injection following onset of EAE. Mice in control and model groups were daily treated with equivalent vehicle. Real-time PCR was used to detect IL-6, IL-17 and TGF-b mRNA levels in the spinal cord and spleen on day 10 and 20 post treatment. ELI?SA was used to examine IL-6, IL-17 and TGF-b expression levels in the brain. Results The neurological deficit scores were significantly different between EAE group and treatment group on 10 and 20 days ( 7.22±1.10 vs. 4.22±0.97 on 10 day, P<0.05;2.89±1.05 vs. 1.33±0.71 on 20 days, P<0.05). The level of of IL-6 in spinal cord was significantly different between EAE group and treatment group on 10 day but not on 20 days (8.54 ± 2.49 vs. 14.07 ± 3.19 on 10 days, P<0.05;2.89±1.05 vs. 1.33±0.71, P>0.05). The level of IL-17 in spinal cord were significantly different between EAE group and treatment group on 10 and 20 days ( 25.50 ± 11.77 vs. 47.59 ± 8.01 on 10 day, P<0.05;10.02 ± 2.99 vs. 20.14 ± 4.57 on 20 day, P<0.05). The level of TGF-βin spinal cord were significantly different between EAE group and treatment group on 10 and 20 days (0.74±0.11 vs. 0.04±0.01 on 10 day, P<0.05;1.49±0.30 vs. 0.59±0.23 on 20 day, P<0.05). The level of IL-6 in brain were significantly different between EAE group and treatment group on 10 and 20 days (44.58 ± 6.94 vs. 57.46±7.32 on 10 day, P<0.05;35.85±9.58 vs. 51.29±2.09 on 20 day, P<0.05). The level of IL-17 in brain were signifi?cantly different between EAE group and treatment group on 10 and 20 days (515.90±63.10 vs. 899.89±242.39 on 10 day, P<0.05;437.71 ± 137.35 vs. 730.32 ± 139.94 on 20 day, P<0.05). The level of TGF-βin brain were significantly different between EAE group and treatment group on 10 and 20 days (170.88±24.69 vs. 132.85±20.27 on 10 day, P<0.05;229.59± 32.10 vs. 172.58 ± 16.94 on 20 day, P<0.05). The level of IL-6 in spleen was significantly different between EAE group and treatment group on 10 day but not on 20 day (6.51±1.29 vs. 19.76±5.85 on 10 day, P<0.05; 3.13±0.78 vs. 4.14±0.58 on 20 day, P>0.05). The level of IL-17 in spleen on 10 day was significantly different between EAE group and treatment group on 10 and 20 days (2.08±1.19 vs. 7.50±1.31 on 10 day, P<0.05;0.67±0.41 vs. 2.75±0.44 on 20 day, P<0.05). The level of TGF-βin spleen was significantly different between EAE group and treatment group on 10 and 20 days (0.61 ± 0.16 vs. 0.08±0.02 on 10 day, P<0.05;1.77±0.39 vs. 0.50±0.33 on 20 day, P<0.05) . Conclusions NDGA can ameliorate the development of EAE by reducing IL-6, IL-17 production and increasing TGF-βproduction.