芪蛎消瘿汤对NOD.H-2h4小鼠甲状腺IL-23/IL-17轴的调控影响

Mediating effects of Qili Xiaoying Decoction on the inflammatory axis of IL-23/IL-17 in NOD.H-2h4 Mice

目的:从辅助性T细胞17(Th17)细胞分化的调控角度,探讨芪蛎消瘿汤治疗自身免疫甲状腺炎(AIT)的作用机制。方法:雌性8周龄NOD.H-2h4小鼠,随机分成4组。正常组(NC)饲以双蒸水,模型组(MC)、常规剂量中药组(ZY1)和3倍剂量中药组(ZY3)自由饮用含0.05%碘化钠水8周成模后,再灌胃给药8周。RIA法测血清TT3、TT4,IRMA法测血清TSH;光镜下观察甲状腺细胞形态;免疫组化法及RT-PCR法分别检测甲状腺IL-6、TGF-β表达;Western Blot法检测甲状腺RORγt、RORα、STAT3蛋白表达。结果:各组间血清TT3、TT4及TSH值比较无统计学差异;与NC组比较,MC组AIT的发生率,IL-6、TGF-βmRNA及蛋白表达,RORγt、RORα、STAT3表达显著升高(P<0.01);与MC组比较,ZY1组IL-6、TGF-β、RORα、STAT3、RORγt表达均明显下降(P<0.01),ZY3组IL-6、TGF-β的整合光密度值及RORα、STAT3表达明显下降(P<0.01),ZY3组RORγt表达下降(P<0.05)。结论:芪蛎消瘿汤可能通过下调IL-6、TGF-β、RORγt、RORα、STAT3的表达而调控IL-23/IL-17炎症轴起到治疗AIT的作用。

Objective： To explore potential mechanism of Qili Xiaoying decoction on mediating Th17 cells differentiation in NOD.H-2h4 mice. Method： 8 w-old NOD.H-2h4 female mice were randomly divided into four groups. Mice in normal control group（NC） received sterile water, while mice in all of the model control group（MC）, the Qili Xiaoying decoction group（ZY1） and the 3 times dose of Qili Xiaoying decoction group （ZY3） were administrated with 0.05% NaI for 8 weeks. After 8 weeks of decoction or sterile water gavages, the mice were sacrificed. The serum TT3 and TT4 were detected by RIA method, the serum TSH by IRMA; thyrocyte morphology was observed under optical microscopy; imrnunohistochemistry staining and RT-PCR were conducted to analyze the expression of IL-6 and TGF-13 respectively, and expressions of RORγ, RORα, STAT3 were analyzed by western blot. Result： No obvious difference among every group on serum TT3,TT4 and TSH; Compared to the NC group, the occurrence ratio of AIT, the expressions of IL-6, TGF-β, RORγt, ROR α and STAT3 in the MC increased markedly （P〈0.01）; Compared to the MC group, the expressions of IL-6, TGF-β, ROR α, STAT3, RORγt in both ZY1 and ZY3 significantly decreased （P〈0.05）. Conclusions： The therapeutic mechanism of Qili Xiaoying decoction on AIT may be related to mediate the inflammatory axis of IL-23/IL-17 by down-regulating the expressions of IL-6, TGF-β, RORyt, RORct, STAT3.