Chinese Medical Sciences Journal ›› 2020, Vol. 35 ›› Issue (3): 215-225.doi: 10.24920/003594

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二氢睾酮对Graves病小鼠模型共刺激分子表达的影响

刘莲叶1,2,施秉银1,赵凤仪1,侯鹏1,刘淑1,刘晓梅1,3,伍丽萍1,*()   

  1. 1西安交通大学第一附属医院内分泌科,西安710061
    2渭南市中心医院内分泌科,渭南714000
    3南京医科大学附属南京医院(南京市第一医院)内分泌科,南京 210001
  • 收稿日期:2019-10-25 出版日期:2020-09-30 发布日期:2020-09-25
  • 通讯作者: 伍丽萍 E-mail:plwzw999@163.com

Effect of Dihydrotestosterone on CostimulatoryMolecules in a Mouse Model of Graves’ Disease

Liu Lianye1,2,Shi Bingyin1,Zhao Fengyi1,Hou Peng1,Liu Shu1,Liu Xiaomei1,3,Wu Liping1,*()   

  1. 1Department of Endocrinology, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061, China
    2Department of Endocrinology, Weinan Central Hospital, Weinan, Shaanxi 714000, China
    3Department of Endocrinology, Nanjing First Hospital, Nanjing Medical University, Nanjing 210001, China
  • Received:2019-10-25 Online:2020-09-30 Published:2020-09-25
  • Contact: Wu Liping E-mail:plwzw999@163.com

摘要:

目的 Graves病是最常见的自身免疫性甲状腺疾病,男性与女性的患病率和临床表现不尽相同。共刺激分子是调节自身免疫反应的重要因子。本研究旨在探讨二氢睾酮(DHT)治疗对自身免疫性Graves病BALB/c模型小鼠表达抑制性共刺激分子的影响。
方法 采用以腺病毒为载体的促甲状腺激素受体A亚单位免疫雌性BALB/c小鼠,3次免疫后建立Graves病模型。在第一次免疫接种前一周,在模型鼠颈部皮下植入含有3种不同剂量DHT及对应安慰剂的缓释剂。第三次免疫4周后处死小鼠,切除脾脏和胸腺。采用放射免疫法分析小鼠血清总甲状腺素和游离甲状腺素水平,RT-PCR测定小鼠脾脏和胸腺淋巴细胞中共刺激分子的表达,流式细胞术分析小鼠脾脏细胞CD4+ T细胞百分比。
结果 DHT能降低模型小鼠血清总甲状腺素和游离甲状腺素水平。与安慰剂组相比,5mg DHT治疗组小鼠脾脏表达程序性凋亡因子1显著增加(0.635±0.296比0.327±0.212;t=2.714,P=0.014);小鼠脾脏(1.004±0.338比0.646±0.314;t=2.205,P=0.022)与胸腺(0.263±0.127比0.120±0.076;t=3.221,P=0.004)表达TIM-3也显著增加;脾脏CD4+ T细胞百分比显著下降(19.90%±3.985%比24.05%±2.587%;t=2.804,P<0.05)。DHT 5mg治疗组小鼠脾脏TIM-3的表达量与血清总甲状腺素水平(r=-0.7106,P=0.014)和游离甲状腺素水平(r=-0.6542,P=0.029)呈负相关。
结论 DHT能改善自身免疫性Graves病模型小鼠病情,其作用可能通过上调程序性凋亡因子1和TIM-3表达以及抑制小鼠脾脏CD4+ T细胞生成实现。

关键词: Graves病, 二氢睾酮, 共刺激分子, 抑制性

Abstract:

Objective Graves’ disease is the most common autoimmune thyroid disease and its prevalence and clinical manifestations are disparate between females and males. Costimulatory molecules play an essential role in regulating autoimmune responses. The objective of this study was to determine if expression of inhibitory molecules was correlated with treatment by dihydrotestosterone (DHT) in an in vivo BALB/c mouse model of experimental autoimmune Graves’ disease.
Methods Female BALB/c mice were immunized three times with thyroid stimulating hormone receptor A-subunit encoded by adenovirus to establish a Graves’ disease model. Three different doses of DHT or a matching placebo were administered by implantation of slow-release pellets a week before the first immunization. Four weeks after the third immunization, the mice were euthanatized, and then the spleen and thymus were removed. Total thyroxine and free thyroxine levels in serum of mice were detected using a radioimmunoassay kit. Real-time polymerase chain reaction was performed to estimate the expression of costimulatory molecules in lymphocytes from the spleen and thymus. Flow cytometry was used to analyze the percentage of CD4+ T cells in splenic lymphocytes. Quantitative data were compared with unpaired t-tests. Correlation between two variables was analyzed using Analysis of Variance.
Results Treatment with DHT can dramatically reduce total thyroxine and free thyroxine levels. Higher expression of programmed death-1 was found in the spleen of Graves’ disease mice receiving 5 mg of DHT treatment (0.635±0.296 vs. 0.327±0.212; t=2.714, P=0.014), similarly, T-cell immunoglobulin domain and mucin domain 3 (TIM-3) in both the spleen (1.004±0.338 vs. 0.646±0.314; t=2.205, P=0.022) and the thymus (0.263±0.127 vs. 0.120±0.076; t=3.221, P=0.004) also increased after 5 mg of DHT treatment compared with the parallel placebo model mice. Moreover, the percentage of CD4+ T cells declined in the splenic lymphocytes of Graves’ disease mice treated with 5 mg of DHT (19.90%±3.985% vs. 24.05%±2.587%; t=2.804, P=0.012). A significant negative association was observed between expression of TIM-3 in the spleen and serum levels of total thyroxine (r=-0.7106, P=0.014) as well as free thyroxine (r=-0.6542, P=0.029).
Conclusion This study demonstrates that DHT can ameliorate experimental autoimmune Graves’ disease, which may occur by up-regulating expression of programmed death-1 and TIM-3 and inhibiting development of CD4+ T cells.

Key words: Graves’ disease, 5α-dihydrotestosterone, costimulatory molecules, inhibitory molecules

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