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目的:探讨黄连温胆汤加减方对大鼠实验性自身免疫性神经炎(EAN)的影响及其机制。方法:应用周围神经髓鞘抗原(P257-81多肽)与完全弗氏佐剂的混合液免疫Lewis大鼠,建立EAN动物模型并随机分为黄连温胆汤加减方高剂量组(HL高组)、黄连温胆汤加减方低剂量组(HL低组)、EAN模型组、正常组。HL高组和HL低组于免疫当天至第15天每天固定时段灌胃给予黄连温胆汤加减方浓煎剂1次,剂量分别为26.89,6.79 g·kg-1·d-1。观察各组发病情况,坐骨神经中T细胞、巨噬细胞浸润及组织病理学变化,检测外周血中17型辅助T细胞(Th17细胞)和叉头状转录因子3阳性调节T细胞(Foxp3+Treg细胞)比例和淋巴结中肿瘤坏死因子-α(TNF-α)、干扰素-γ(IFN-γ)、白细胞介素-6(IL-6)、白细胞介素-17(IL-17)mRNA水平。结果:HL高组的最初发病时间为(11.92±1.38)d,迟于EAN组(P<0.05),其高峰期临床评分为(2.41±1.09),髓鞘脱失的组织学评分为(0.58±0.51),均显著低于EAN组(P<0.05);HL高组和HL低组T细胞、巨噬细胞浸润明显减少,外周血中Th17细胞比例(1.88±0.47)%,(2.71±0.39)%较EAN组显著减少(P<0.05),Foxp3+Treg细胞比例(15.06±1.35)%,(9.18±1.00)%较EAN组显著增加(P<0.05),淋巴结中促炎细胞因子TNF-α,IFN-γ,IL-6,IL-17mRNA表达HL高组分别为(0.38±0.02)%,(0.45±0.08)%,(0.34±0.13)%,(0.63±0.17)%;HL低组分别为(0.45±0.03)%,(0.60±0.05)%,(0.74±0.12)%,(1.04±0.17)%与EAN组比较明显下降(P<0.05)。结论:黄连温胆汤加减方能够抑制EAN大鼠的自身免疫反应,对EAN有治疗作用。
Objective: To investigate the effect of Huanglian Wendan Decoction on experimental autoimmune neuritis (EAN) in rats and its mechanism. Methods: Lewis rats were immunized with a mixed solution of peripheral nerve myelin antigen (P257-81) and complete Freund’s adjuvant to establish animal model of EAN and randomly divided into high-dose group (HL high-dose group) , Huanglian Wendan Decoction low dose group (HL low group), EAN model group, normal group. HL high group and HL low group from day of immunization to day 15 fixed period of time by intragastric administration of Huanglianwendan decoction decoction decoction once, the doses were 26.89,6.79 g · kg-1 · d-1. To observe the incidence of each group, the changes of T cells and macrophages in the sciatic nerve and histopathological changes, and to detect the expression of 17 type helper T cells (Th17 cells) and forkhead transcription factor 3 (Tregs) in peripheral blood (Foxp3 + Treg cells ) And the levels of tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-6 (IL-6) and interleukin-17 (IL- Results: The initial onset time of HL group was (11.92 ± 1.38) d, which was later than that of EAN group (P <0.05). The peak score of HL group was (2.41 ± 1.09) and the histological score of myelin degeneration was (0.58 ± 0.51) were significantly lower than those in EAN group (P <0.05). The infiltration of T cells and macrophages in HL group and HL group were significantly decreased, the proportion of Th17 cells in peripheral blood was (1.88 ± 0.47)%, (2.71 ± 0.39) )% In EAN group was significantly lower than that in EAN group (P <0.05). The proportion of Foxp3 + Treg cells (15.06 ± 1.35)%, (9.18 ± 1.00)% in EAN group was significantly higher than that in EAN group (0.38 ± 0.02)%, (0.45 ± 0.08)%, (0.34 ± 0.13)%, (0.63 ± 0.17)%, respectively. HL low group (0.45 ± 0.03)%, (0.60 ± 0.05)%, (0.74 ± 0.12)% and (1.04 ± 0.17)%, respectively, compared with EAN group (P <0.05). Conclusion: Huanglian Wendan Decoction can inhibit the autoimmune response of EAN rats and has a therapeutic effect on EAN.