Effect of Qili Xiaoying Decoction on the CD4T Cell Subsets in NOD. H-2 Mice with Autoimmune Thyroiditis

YANG Xiao; GAO Tian-shu; ZHOU Xi-yu; XU Jia; QU Jin-qiao

doi:10.11653/syfj2014060149

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Effect of Qili Xiaoying Decoction on the CD4T Cell Subsets in NOD. H-2 Mice with Autoimmune Thyroiditis

更新时间：2024-01-04

- Effect of Qili Xiaoying Decoction on the CD4T Cell Subsets in NOD. H-2 Mice with Autoimmune Thyroiditis
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 20, Issue 6, Pages: 149-153(2014)
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- DOI：10.11653/syfj2014060149
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- Published：2014
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YANG Xiao, GAO Tian-shu, ZHOU Xi-yu, et al. Effect of Qili Xiaoying Decoction on the CD4T Cell Subsets in NOD. H-2 Mice with Autoimmune Thyroiditis[J]. Chinese journal of experimental traditional medical formulae, 2014, 20(6): 149-153.
DOI：

YANG Xiao, GAO Tian-shu, ZHOU Xi-yu, et al. Effect of Qili Xiaoying Decoction on the CD4T Cell Subsets in NOD. H-2 Mice with Autoimmune Thyroiditis[J]. Chinese journal of experimental traditional medical formulae, 2014, 20(6): 149-153. DOI： 10.11653/syfj2014060149.

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-4 mRNA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。

Abstract

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.②Lymphocytic 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 Th1

Th2

Treg by up-regulating the expression of INF-γ

IL-4 and Foxp3.

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