【摘要】 目的 本研究旨在通过比较在两种表达人促甲状腺激素受体(thyrotropin receptor, TSHR)A亚单位基因的质粒载体在电穿孔介导下诱导Graves病动物模型的效果，为探索Graves病防治方法提供更为有效的研究工具。方法 构建表达TSHR A亚单位的两种质粒pcDNA3.1-TSHR A和pTriEx1.1-TSHR A,通过对BALB/c小鼠肌内注射并立即电穿孔的方式诱导Graves病，每3周1次，共4次。对照组小鼠使用PBS。在第2次电穿孔后1周采血测定血清TSHR抗体(thyrotropin receptor antibodies, TRAb)。末次电穿孔后3周对小鼠进行心脏超声检查。末次电穿孔后4周处死小鼠，收集血液、甲状腺及眼眶组织，测定血清总甲状腺素(total thyroxine, TT4)并进行组织病理学检查。结果 pcDNA3.1-TSHR A组(n=15)和pTriEx1.1-TSHR A组(n=13)小鼠血清TRAb平均值分别为(6.9±2.0)U/L和(7.5±2.2)U/L。后者显著高于对照组(4.9±0.5)U/L (P=0.033)。pcDNA3.1-TSHR A组和pTriEx1.1-TSHR A组小鼠血清TT4平均值分别为(41.4±23.8)ng/mL和(63.2±53.7)ng/mL,均高于对照组(20.2±4.0)ng/mL (P<0.01)。甲状腺病理显示模型组小鼠甲状腺滤泡上皮增生并且有T细胞浸润。心脏超声显示pTriEx1.1-TSHR A组的左心室质量高于对照组(P=0.007)和pcDNA3.1-TSHR A组(P=0.012)。眼眶病理显示模型组小鼠的眼外肌中存在纤维化改变。结论 表达TSHR A亚单位的pcDNA3.1和pTriEx1.1均能通过电穿孔法成功诱导Graves病小鼠模型，且诱导甲状腺功能亢进和甲亢眼病的效能相近。pTriEx1.1-TSHR A诱导甲状腺毒症性心脏病的效能优于pcDNA3.1-TSHR A。更多还原

【Abstract】 Objective The purpose of this study was to provide a more effective method for researching the prevention and treatment of Graves’ disease by comparing the effects of two plasmid vectors expressing the human thyrotropin receptor(TSHR) A subunit gene in inducing an animal model of Graves’ disease via electroporation. Methods Plasmids pcDNA3.1-THSR A, and pTriEx1.1-THSR A expressing the TSHR A subunit were constructed and used to induce Graves’ disease by intramuscular injection with immediate electroporation once every 3 weeks for a total of 4 times. Mice in the control group were injected with PBS. One week after the second electroporation, blood was collected to measure serum thyrotropin receptor antibody(TRAb). Three weeks following the last electroporation, echocardiography was performed on the mice. Mice were sacrificed 4 weeks after the last electroporation; blood, thyroid, and orbital tissues were collected; serum total thyroxine(TT4) was measured; and histological examination was performed. Results The average concentrations of serum TRAb in the pcDNA3.1-TSHR A group(n = 15) and the pTriEx1.1-TSHR A group(n = 13) were(6.9 ± 2.0) U/L and(7.5 ± 2.2) U/L, respectively. The latter was significantly higher than that in the control group(4.9 ± 0.5) U/L(P = 0.033). The average concentrations of serum TT4 in the pcDNA3.1-TSHR A group and pTriEx1.1-TSHR A group were(41.4 ± 23.8) ng/mL and(63.2 ± 53.7) ng/mL, respectively, both higher than that in the control group:(20.2 ± 4.0) ng/mL(P < 0.01). Thyroid pathology showed thyroid follicular epithelial hyperplasia with T-cell infiltration in the model group. Echocardiography showed that the left ventricle mass in the pTriEx1.1-TSHR A group was higher than those in the control group(P = 0.007) and pcDNA3.1-TSHR A group(P = 0.012). Orbital pathology showed fibrotic changes in the extraocular muscles of mice in the model groups. Conclusions Both pcDNA3.1 and pTriEx1.1 expressing the TSHR A subunit were able to induce Graves’ disease in mice by electroporation, and the efficiency of the two plasmids in inducing hyperthyroidism and Graves’ ophthalmopathy was similar. The efficiency of pTriEx1.1-TSHR A in inducing thyrotoxic heart disease was better than that of pcDNA3.1-TSHR A.更多还原