Xia Nan, Ye Xiaozhen, Hu Xiaohao, Song Shiyu, Xu Hui, Niu Mengyuan, Wang Hongwei, Wang Jian
Department of Endocrinology, Jingling Hospital, Medical School of Nanjing University, Nanjing, P.R. China.
Center for Translational Medicine and Jiangsu Key Laboratory of Molecular Medicine, Medical School of Nanjing University, Nanjing, P.R. China.
PLoS One. 2017 Mar 20;12(3):e0174260. doi: 10.1371/journal.pone.0174260. eCollection 2017.
Graves' disease is the most common form of autoimmune thyroid disorder, characterized by hyperthyroidism due to circulating autoantibodies. To address the pathological features and establish a therapeutic approach of this disease, an animal model carrying the phenotype of Graves' disease (GD) in concert with Graves' Ophthalmopathy (GO) will be very important. However, there are no ideal animal models that are currently available. The aim of the present study is to establish an animal model of GD and GO disease, and its pathological features were further characterized.
A recombinant plasmid pcDNA3.1- T289 was constructed by inserting the TSHR A-subunit gene into the expression vector pcDNA3.1, and genetic immunization was successfully performed by intramuscular injection of the plasmid pcDNA3.1-T289 on female 8-week-old BALB/c mice. Each injection was immediately followed by in vivo electroporation using ECM830 square wave electroporator. Morphological changes of the eyes were examined using 7.0T MRI scanner. Levels of serum T4 and TSHR antibodies (TRAb) were assessed by ELISA. The pathological changes of the thyroid and orbital tissues were examined by histological staining such as H&E staining and Alcian blue staining.
More than 90% of the immunized mice spontaneously developed goiter, and about 80% of the immunized mice manifested increased serum T4 and TRAb levels, combined with hypertrophy and hyperplasia of thyroid follicles. A significantly increased synthesis of hyaluronic acid was detected in in the immunized mice compared with the control groups.
We have successfully established an animal model manifesting Graves' hyperthyroidism and ophthalmopathy, which provides a useful tool for future study of the pathological features and the development of novel therapies of the diseases.
格雷夫斯病是自身免疫性甲状腺疾病最常见的形式，其特征是由于循环自身抗体导致甲状腺功能亢进。为了研究这种疾病的病理特征并建立治疗方法，构建一种同时具有格雷夫斯病（GD）和格雷夫斯眼病（GO）表型的动物模型将非常重要。然而，目前尚无理想的动物模型。本研究的目的是建立GD和GO疾病的动物模型，并进一步表征其病理特征。
通过将促甲状腺激素受体A亚基基因插入表达载体pcDNA3.1构建重组质粒pcDNA3.1-T289，并通过对8周龄雌性BALB/c小鼠肌肉注射质粒pcDNA3.1-T289成功进行基因免疫。每次注射后立即使用ECM830方波电穿孔仪进行体内电穿孔。使用7.0T MRI扫描仪检查眼睛的形态变化。通过ELISA评估血清T4和促甲状腺激素受体抗体（TRAb）水平。通过苏木精-伊红染色和阿尔辛蓝染色等组织学染色检查甲状腺和眼眶组织的病理变化。
超过90%的免疫小鼠自发出现甲状腺肿大，约80%的免疫小鼠血清T4和TRAb水平升高，同时伴有甲状腺滤泡肥大和增生。与对照组相比，免疫小鼠体内透明质酸的合成显著增加。
我们成功建立了一种表现为格雷夫斯甲状腺功能亢进和眼病的动物模型，为今后研究该疾病的病理特征和开发新疗法提供了有用的工具。
