Establishment of BALB/c mice models for Graves disease

Abstract

Objective To establish stable Graves disease (GD) mice models with immunization and electroporation (EP).Methods Fifty mice were divided into 3 groups by random number table method:experimental group (n =30),control group (n =10),blank group (n =10).Recombinant plasmid pcDNA3.1/hTSHR268 was constructed and injected to bilateral gastrocnemius in experimental group mice on the 1st,4th,7th and 10th week.The same volume of normal saline was injected in the control group and blank group at the same time.Both experimental group and control group were subjected to EP at the same time and the same location to enhance immunization.Serum T4 was tested with radioimmunoassay.TRAb N-terminal (TRAb N) and TRAb C-terminal (TRAb C) antibodies were tested with ELISA.Whole body 99TcmO4-imaging was performed and then thyroid morphology and pathology were investigated.Data were analyzed by one-way analysis of variance and the least significant difference (LSD) t test.Results GD BALB/c mice models were built successfully (80％,24/30).Serum T4 increased from (16.06±5.16) nmol/L at the basic level to(95.04±68.92) nmol/L on the 12th week(F=18.906,t=-5.598,P＜0.05).Serum TRAb N antibody increased from (0.006±0.002) U/L at the basic level to (0.251±0.110) U/L on the 12th week(F=47.491,t=-10.869,P＜0.05).Serum TRAb C antibody increased from (11.176±2.635)×103 arbitrary unit (AU)/L at the basic level to (46.395±22.001)× 103 AU/L on the 12th week(F=14.642,t =-7.787,P＜0.05).On the 18th week serum T4,TRAb N and TRAb C decreased to (36.64±23.68) nmol/L,(0.094±0.053) U/L and (24.456±6.725)× 103 AU/L respectively,which were still higher than those preimmune levels(t=-4.161,-8.085,-9.008,all P＜0.05).There were no significant change of T4,TRAb N and TRAb C in the control group and blank group.After 4 times of immunization,the 99TcmO4-uptake by thyroids in immunized mice increased.The thyroid glands of immunized mice showed enlargement.Microscope examination showed that there were lymphocytes infiltration,colloid decrease and epithelial cell proliferation in thyroids of immunized mice.Conclusion GD mice models were successfully established by injecting recombinant plasmid pcDNA3.1/hTSHR268 and EP. Key words: Graves disease; Models, animal; Mice