芪蛎消瘿汤对NOD.H-2h4小鼠甲状腺CD4＋T细胞亚群影响的实验研究  

Effect of Qili Xiaoying Decoction on the CD4^+ T Cell Subsets in NOD. H-2^(h4) Mice with Autoimmune Thyroiditis

作　　者：杨潇[1,2] 高天舒[3] 周喜玉[2] 徐佳[2] 曲金桥 

机构地区：[1]辽宁中医药大学第二附属医院,沈阳110034 [2]辽宁中医药大学,沈阳110032 [3]辽宁中医药大学附属医院,沈阳110032

出　　处：《中国实验方剂学杂志》2014年第6期149-153,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：沈阳市科技局计划(F10-205-1-31);辽宁省教育厅优秀人才计划(LR201026);教育部博士点基金项目(20092133110004)

摘　　要：目的:从CD4+T细胞亚群角度,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:60只SPF级NOD.H-2h4小鼠,随机分成以下4组:正常(NC)组、模型(MC)组、常规剂量芪蛎消瘿汤组(ZY1,20.8 g·kg-1)、和3倍剂量芪蛎消瘿汤组(ZY3,62.4 g·kg-1),正常组饲以双蒸水,其余各组自由饮用含0.05%碘化钠水8周,8周后自发形成AIT动物模型,再给处理因素8周后处死小鼠,留取各组NOD.H-2h4小鼠血清、甲状腺。放免法(RIA法)测小鼠血清总三碘甲状腺原氨酸(TT3)、总甲状腺素(TT4),固相化学发光酶免疫法(IRMA法)测定小鼠血清促甲状腺激素(TSH);光镜下观察甲状腺细胞形态;免疫组化法测定小鼠甲状腺γ-干扰素(INF-γ)、白细胞介素-4(IL-4)蛋白表达;RT-PCR法检测小鼠甲状腺INF-γ,IL-4mRNA表达;Western blot法测定小鼠甲状腺组织叉头翼状螺旋转录因子(Foxp3)蛋白表达。结果:各组间血清TT3,TT4及TSH值比较无明显差异。HE染色显示MC组甲状腺组织淋巴细胞浸润,MC组AIT的发生率明显高于NC组(P<0.01)。MC组INF-γ,IL-4 mRNA及蛋白表达较NC组显著下降(P<0.01);与MC组比较,ZY1,ZY3组INF-γ,IL-4的蛋白表达升高(分别P<0.01,P<0.05);且ZY1组,ZY3组INF-γ,IL-4mRNA表达显著升高(P<0.01)。MC组甲状腺组织Foxp3的蛋白表达量较NC组明显下降(P<0.01);ZY1,ZY3组Foxp3表达较MC组明显升高(P<0.01)。结论:芪蛎消瘿汤治疗AIT的机制可能与上调INF-γ,IL-4,Foxp3的表达,影响NOD.H-2h4小鼠Th1,Th2,Treg细胞亚群水平相关。Objective：To explore the mechanism of Qili Xiaoying decoction on NOD.H-2h4 mice with autoimmune thyroiditis （AIT） in view of CD4 ＋T cell subsets.Method：Sixty SPF NOD.H-2h4 mice were randomly divided into four groups,the normal control group （NC）,the model control group （MC）,the Qili Xiaoying decoction group （ZY1,20.8 g ·kg-1） and the 3 times dose of Qili Xiaoying decoction group （ZY3,62.4 g·kg-1）.The NC group received sterile water,and the others were administrated with 0.05％ NaI for 8 weeks to induce AIT.All mice were sacrificed at the time point of giving treatment factors for 8 weeks.The serum triiodothyronine （TT3） and thyroxine （TT4） were detected by RIA method,the serum thyroid-stimulating hormone （TSH） by IRMA.Thyrocyte morphology was observed under optical microscopy.Immunohistochemistry staining was conducted to analyze the expression of INF-γ and IL-4.The expression of INF-γ mRNA and IL-4 mRNA in NOD.H-2h4 mice thyroid was detected by Real-time （RT）-PCR,and the Foxp3 expressions in the NOD.H-2h4 mice thyroid were determined by Western blot.Result：① No apparent differences among every group in serum TT3,TT4 and TSH were found.② Lymphocytic infiltration could be found by HE staining,AIT occurrence rate of MC was obviously higher than the NC （P ＜ 0.01）.③ Compared with the NC group,expression of INF-γ mRNA,IL-4 mRNA and protein in the MC decreased markedly （P ＜ 0.01） ; compared to the MC,the protein expression of INF-γ and IL-4 both in ZY1 and ZY3 were increased （respectively P ＜ 0.01,P ＜0.05）.The expression of INF-γ mRNA and IL-4 mRNA in ZY1 and ZY3 were also increased remarkably （P ＜ 0.01）.④ The protein expressions of Foxp3 in the MC declined markedly compared with the NC （P ＜ 0.01） ; the Foxp3 expression in ZY1 and ZY3 ascended remarkably compared with the MC （P ＜ 0.01）.Conclusion：The therapeutic mechanism of Qili Xiaoying decoction on NOD.H-2h4 with AIT may be related to influencing the cell subsets of T

关 键 词：芪蛎消瘿汤 NOD H-2h4小鼠 自身免疫甲状腺炎 Γ-干扰素 白细胞介素-4 叉头翼状螺旋转录因子 

分 类 号：R285.5[医药卫生—中药学]