活化的脾细胞联合基因免疫共同诱导甲状腺相关性眼病动物模型的研究
Study of activated spleen cells combined with gene immunity to induce thyroid associated ophthalmopathy in animal model
孙宝琪 1孙增刚 2刘凯峰 3王晓琳 4史明明 5张艳辉 3张金梅 6邵光东5
作者信息
- 1. 山东第二医科大学附属医院眼科中心,山东潍坊 261031
- 2. 山东第二医科大学麻醉学院
- 3. 山东第二医科大学医学影像学院
- 4. 潍坊市中医院病理科
- 5. 潍坊市中医院甲状腺乳腺外科
- 6. 潍坊市中医院内分泌科
- 折叠
摘要
目的 应用表达人促甲状腺素受体(hTSHR)重组质粒介导的基因免疫,联合pcDNA3.1/hTSHR基因免疫活化的脾细胞,共同免疫BALB/c小鼠,构建甲状腺相关眼病(TAO)小鼠模型.方法 选择BALB/c雌性小鼠35只,随机分为空白质粒组(A组)、基因免疫组(B组)和活化脾细胞联合基因共同免疫组(C组),每组10只.分别给予空白质粒pcDNA3.1、重组质粒pcDNA3.1/hTSHR、重组质粒pcDNA3.1/hTSHR+重组质粒pcD-NA3.1/hTSHR免疫活化脾细胞干预处理.所有小鼠均于免疫第22周行眼眶MRI检查后处死,应用光学显微镜和透射电子显微镜观察眼眶组织和甲状腺组织,酶联免疫吸附法检测血清T3、T4、TSH、TPOAb及TRAb的表达水平.结果 外观:与A组相比,50％的B组小鼠和80％的C组小鼠表现出不同程度的眼球突出,且C组小鼠眼球突出程度更明显.影像学:B组和C组小鼠眼外肌体积较A组增大,且以C组增大更为明显.组织病理学形态:A组小鼠眼眶及甲状腺组织均未见异常改变;B组小鼠眼眶肌纤维变性断裂、粗细不均,纤维结缔组织,肌间少量胶原纤维增生,甲状腺滤泡间及纤维中见少量淋巴细胞浸润;C组小鼠眼眶肌纤维变性断裂、粗细不均、横纹模糊、肌浆凝聚,纤维结缔组织明显水肿伴粘液变性,肌间大量胶原纤维及脂肪组织增生,甲状腺滤泡间及纤维中见少量淋巴细胞浸润.血清学指标:与A组相比,B、C组均出现T3、T4、TRAb升高及TSH降低,且以C组改变更为明显;三组之间TPOAb数值相比无明显差异.结论 本研究成功建立了一种改良的TAO动物模型,为进一步研究该病的病理特征和开发新的治疗方法提供了有用的工具.
Abstract
Objective Constructing a Thyroid Associated Ophthalmopathy(TAO)mouse model by co-im-munizing BALB/c mice with gene immunization containing human thyroid stimulating hormone receptor(hTSHR)and splenocytes immunoactivated by the pcDNA3.1/hTSHR gene.Methods Thirty five female BALB/c mice were randomly divided into three groups.10 mice(Group A)received pcDNA3.1,10 mice(Group B)received pcDNA3.1/hTSHR and 10 mice(Group C)received the same dose of pcDNA3.1/hTSHR and splenic cells immunologically activated by pcD-NA3.1/hTSHR.All mice were sacrificed after orbital MRI examination at week 22 of immunization.Orbital tissue and thyroid tissue were observed by optical microscopy and transmission electron microscopy,and serum T3,T4,TSH,TPOAb and TRAb expression levels were detected by enzymelinked immunosorbent assay.Results 50％of mice in Group B ex-hibited varying degrees of exophthalmos and conjunctiva redness,and MRI imaging also showed an increase in extraocu-lar muscle volume compared to Group A.Imaging:the extraocular volume of mice in group B and group C was larger than that in group A,and the increase was more obvious in group C.Histopathological:No abnormal changes were observed in orbital and thyroid tissue of group A mice.In group B,orbital muscle fibers were degenerated and broken,with uneven thickness,fibrous connective tissue,a small amount of collagen fiber hyperplasia between muscles,and a small amount of lymphocyte infiltration between thyroid follicles and fibers.In group C,the orbital muscle fibers were degenerated and broken,the thickness was uneven,the transverse lines were blurred,the sarcoplasmic aggregation,the fibrous connective tissue edema with mucinous degeneration,the significant collagen and adipocyte proliferation in the muscle,and a small amount of lymphocyte infiltration in the thyroid follicles and fibers.In terms of serology,compared with group A,T3,T4,TRAb were increased and TSH was decreased in group B and C,and the changes were more obvious in group C.There was no significant difference in TPOAb values among the three groups.Conclusion We have successfully established an animal model of TAO,providing useful tools for further studying the pathological characteristics of the disease and develo-ping new treatment methods.
关键词
甲状腺相关眼病/改良疾病模型/基因免疫/活化脾细胞/双免法Key words
Thyroid associated ophthalmopathy/Improved animal model/Genetic immunization/Activated splenocytes/Dual immunity method引用本文复制引用
出版年
2024
NETL
NSTL
万方数据