用hTSHR活化脾细胞免疫小鼠建立Graves病动物模型的研究  

Model establishment of Graves’disease by immunizing mice with hTSHR-activated splenocytes

作　　者：何云[1] 刘纯[1] 张徽[2] 张琳[1] 詹升华[1] 李秀娟[1] 随华[1] 

机构地区：[1]重庆医科大学附属第一医院内分泌科,重庆400016 [2]重庆医科大学基础医学院病理教研室,重庆400016

出　　处：《重庆医科大学学报》2008年第11期1313-1316,共4页Journal of Chongqing Medical University

基　　金：重庆市卫生局重点科研基金资助项目(01-1-104)

摘　　要：目的:用人促甲状腺激素受体活化的脾细胞免疫同源雌性BALB/c小鼠建立Graves病动物模型。方法:第一阶段,将6～8周龄的雌性BALB/c小鼠随机分为2组(a组和b组),分别用重组质粒pcDNA3.1/hTSHR和空质粒pcDNA3.1免疫,第3和6周加强免疫2次。18周时分别处死两组小鼠,分离其脾细胞。第二阶段,分别取a组和b组小鼠的脾细胞免疫同源的两组6～8周龄的雌性BALB/c小鼠(实验组和对照组),4周后光镜和电镜检查甲状腺组织,并测定小鼠血清TT4、TSH和TRAb水平。结果:实验组约44%小鼠甲状腺组织见明显增生,滤泡上皮细胞增多、变大呈立方形,部分突向滤泡腔内;散在免疫细胞侵润。电镜下见滤泡上皮细胞胞浆内有丰富的线粒体和粗面内质网,腔面有发育良好的微绒毛。实验组血清TT4高于对照组(7.13±1.02与6.43±0.57μg/dl)(P<0.05)、TSH水平低于对照组(0.070±0.032与0.098±0.020μIU/ml)(P<0.05)。其中4只小鼠可能发生了甲状腺功能亢进,1只小鼠可能发生了亚临床甲状腺功能亢进。结论:hTSHR活化脾细胞免疫BALB/c小鼠可以有效地产生类似人类Graves病的动物模型。Objective：To establish an animal model of Graves＇disease by immunizing syngeneic BALB/c mice with hTSHR-activated splenocytes. Methods：The female BALB/c mice（6-8 weeks） were randomly divided into 2 groups（group a and group b）, which were immunized with recombinant plasmid pcDNA3.1/hTSHR and blank plasmid pcDNA3.1（±） respectively for 3 times at 3-week intervals.The animals were sacrificed at 18th week and their splenocytes were isolated.Other two groups of female syngeneic BALB/c mice（6-8 weeks） were divided randomly into experimental and control groups, which received splenocytes from group a and group b respectively. Four weeks later, the thyroid glands were removed for histological examination and the sera were determined for Total T4,TSH and TRAb measurements. Results：The thyroid tissues under light microscope displayed diffuse hypercellularity with the follicular epithelia being tall cuboidal cells and reduced colloid in 44% of experimental mice with irregular apical poles protruding into the follicular space; there was minimal immune cell in the interstitium with hyperaemia and edema. Electron microscope examination revealed that the follicular epithelia contained numerous mitochondria and rough endoplasmic reticula,and many long microvilli existed in the luminal surface.The serum level of TF4 in experimental mice was higher than that in control（7.13±1.02 vs 6.43±0.57 μg/dL） （P〈0.05）.The serum level of TSH in experimental mice was lower than that in control（0.070±0.032 vs 0.098±0.020 μIU/ml） （P〈0.05）.Hyperthyroidism probably appeared in 4 mice and subclinical hyperthyroidism in 1 mouse. Conclusion：We have effectively induced an animal model which mimics some characteristics of human Graves＇disease via splenocytes from the animals immunized with recombinant plasmid pcDNA3.1/hTSHR.

关 键 词：GRAVES病 促甲状腺激素受体 脾细胞 免疫 

分 类 号：R581.11[医药卫生—内分泌]