TSHR基因免疫诱导BALB/c小鼠Graves甲亢伴Graves眼病的实验研究  

An experimental study on the induction of Graves′hyperthyroidism with Graves′ophthalmopathy in BALB/c mice by TSHR gene immunization

作　　者：李宁[1] 冯思源 孙磊[3] 郑薇[1] 王深[1] 申一鸣[1] 林晓云[1] 王萱 孟召伟[1] 贾强[1] 谭建[1] 宋其韬[2] Li Ning;Feng Siyuan;Sun Lei;Zheng Wei;Wang Shen;Shen Yiming;Lin Xiaoyun;Wang Xuan;Meng Zhaowei;Jia Qiang;Tan Jian;Song Qitao(Department of Nuclear Medicine, Tianjin Medical University General Hospital, Tianjin 300052, China;Department of Nuclear Medicine, Tianjin Hospital, Tianjin 300211, China;Tianjin Institute of Orthopaedics, Tianjin Hospital, Tianjin 300211, China)

机构地区：[1]天津医科大学总医院核医学科,300052 [2]天津市天津医院核医学科,300211 [3]天津市天津医院骨科研究所,300211

出　　处：《中华内分泌代谢杂志》2019年第5期417-424,共8页Chinese Journal of Endocrinology and Metabolism

基　　金：国家自然科学基金青年科学基金(81601523)。

摘　　要：目的以BALB/c小鼠多次注射促甲状腺素受体(thyrotropin receptor,TSHR)膜外区A亚单位重组质粒(pcDNA3.1/TSHR289)免疫诱导Graves病(Graves′ disease,GD)伴(或)Graves眼病(Graves′ ophthalmopathy,GO)模型。方法实验组小鼠(35只)双侧腓肠肌注射pcDNA3.1/TSHR289并电穿孔;对照组(10只)注射无菌生理盐水并电穿孔;空白组(5只)注射无菌生理盐水。各组小鼠均在第1、4、7、10周免疫,免疫前、每次免疫后2周及末次免疫后5周、8周取血检测血清TT4、TSH、促甲状腺素受体刺激性抗体(thyrotropin receptor stimulating antibody, TSAb)、促甲状腺素受体阻断性抗体(thyrotropin receptor stimulating blocking antibody, TSBAb)。CT平扫评估小鼠眼部变化,99mTcO4-显像评估甲状腺摄取功能,HE染色评估甲状腺和眼眶组织病理变化。组间均数比较采用单因素方差分析。结果GD小鼠2次免疫后即见血清TT4、TSAb和TSBAb均持续显著高于对照组和空白组(分别F=13.781,31.435,36.112,均P<0.01),TSH持续低于对照组和空白组(F=13.966,P<0.01)。4次免疫后GD小鼠甲状腺99mTcO4-摄取能力明显增强,甲状腺肿大,腺内大量淋巴细胞浸润,滤泡胶质稀薄。GO小鼠眼CT扫描见眼外肌增粗肿胀,肌腱及肌附着点未见异常。眼外肌和球后组织可见淋巴细胞、透明质酸和脂肪细胞浸润等病理改变。结论BALB/c小鼠多次肌肉注射pcDNA3.1/TSHR289可成功诱导GD伴(或)GO动物模型。Objective To construct a model of Graves′ disease(GD) with (or) Graves′ ophthalmopathy(GO) in BALB/c mice by immunization with pcDNA3.1/TSHR289. Methods pcDNA3.1/TSHR289 was injected into the bilateral gastrocnemius muscle of 35 model mice and electroporation was immediately performed. 10 control mice were injected with sterile saline and electroporated, while 5 blank mice were injected with sterile saline only. Each group of mice was immunized at 1, 4, 7, and 10 weeks, respectively. Serum total T4, TSH, TSAb, and TSBAb were measured before immunization, 2 weeks after each immunization, as well as 5 and 8 weeks after the last immunization. CT scan was used to evaluate the morphological changes of the eyes of the mice.99mTcO4- imaging was used to measure the thyroid uptake function, and the pathological changes of the thyroid and orbital tissues were evaluated by HE staining. Results After the 2nd time immunization, the serum concentrations of TT4, TSAb and TSBAb in GD mice were significantly higher than those of control and blank groups(F=13.781, 31.435, 36.112, P<0.01, respectively). The TSH continued to be significantly lower than that of control and blank groups(F=13.966, P<0.01). After the 4th time immunizations, the ability of uptaking99mTcO4- in GD mice thyroid was significantly enhanced compared with the control group. The thyroid goiter with a large amount of lymphocyte infiltration, and the thyroid follicle was thin. CT scan of GO mice showed thickening and swelling of the extraocular muscles, and no abnormalities in tendon and muscle attachment points. HE staining showed thickening of extraocular muscle fibers, lymphocyte infiltration of extraocular muscles and orbital tissue, increased hyaluronic acid, and infiltration of fat cells. Conclusion GD or GO model can be successfully induced by multiple intramuscular injection of pcDNA3.1/TSHR289 in BALB/c mice.

关 键 词：格雷夫斯病 格雷夫斯眼病 促甲状腺素受体抗体 小鼠模型 电穿孔 

分 类 号：R581[医药卫生—内分泌]