Effect Of The Pregnant Mice With Graves’ Disease On Thyroid Function In The Offspring

[Objective]1To prepare an animal model by immunizing female Balb/c mice withAdenovirus Expressing TSHR A-subunit(Ad-TSHR-289).2To mate with male micein order to obtain the offspring.3To measure autoantibodies and Thyroxine in allmice’ sera and their mothers’ breast milk.[Methods]Part1:(1) To amplify the Adenovirus Expressing TSHR A-subunit(Ad-TSHR-289) andthe control adenovirus (Ad-β-gal);(2)Female Balb/c mice aged6weeks were dividedrandomly into experimental group and control group, then injected intramuscularlywith Ad-TSHR-289(3.3×10~8PFU/50μlPBS) and the control adenovirus(Ad-β-gal3.3×108PFU/50μlPBS) three times at3weeks intervals.(3) In1wk after the secondinjection, the levels of T4were measured. All mice were sacrificed4wk after the lastimmunization,and then Thyroxine(T4),as well as TSH-receptor antibodies(TRAb)and thyroid,were then examined.(4) T4were tested by radioimmunoassay. TRAbwas tested by electrochemiluminescent immunoassay.(5) Their thyroids wereremoved for histological examination.Part2:(1)(2)The establishment of the Graves’ disease animal model: using the sameway of part1;(3)When the immunization was over, both of groups were mated withmale mice. Experiment group were divided into three groups again. They weresacrificed1d,1wk or3wk after delivery. At the same time, the breast milk from theinfants’ stomach were collected;the same in control group.T4,as well asTRAb,Thyroid peroxidase antibodies(TPOAb), Thyroglobulin antibodies(TGAb) insura and thyroid, were be examined.(4)T4were tested by radio immunoassay. TRAb, TPOAb, TGAb were tested by electrochemiluminescent immunoassay.(5) Theirthyroids were removed for histological examination.Results:Part1:1. Purified recombinant adenovirus with the target gene-TSHR-289was amplified,which size is about800bps.2.7mice in experiment group were suffered with Graves’ hyperthyroidism. Theincidence rate was70%. The levels of serum T4and TRAb in experiment groupwere significantly higher than those of control one(P<0.05).Part2:1、The levels of sera T4and TRAb of mothers1d,1wk and3wk after delivery inexperiment groups were significantly higher than those of control one(P<0.05).22,18and23mothers after1d,1wk and3wk delivery in experiment groups weresuffered with Graves’ hyperthyroidism respectively. The incidence rate respectivelywere73.3%,60%,76.67%.2、We can test TRAb in breast milk from9mice1d after delivery, TPOAb>0.5IU/L in milkfrom15mice, and TGAb in breast milk from4mice. We can test TRAb in breast milkfrom4mice in1wk after delivery, TPOAb in milk from7mice, and TGAb in milk from1mice. TRAb can not be tested in mice’ breast milk in3wk after delivery,2mice could betested TPOAb from their milk, and3mice could be tested TGAb from their milk. The levelsof TPOAb in the breast milk after1d delivery in experiment group were significantlyhigher than those of control group(P<0.05).3、114,129mice aged1wk and3wk in the experiment groups were suffered withGraves’ hyperthyroidism. The incidence rate respectively were63.3%and71.7%.The levels of sera T4, TRAb, TPOAb and TGAb in offspring aged1wk and3wk inexperiment groups were significantly higher than those of control groups(P<0.05). Conclusions:1.An animal model of Graves’ disease is successfully prepared by immunization withAd-TSHR-289.2. TPOAb and TGAb can be affacted by TRAb.3. The higher levels of Autoantibodies could result in neonatal thyroid disease bytransfer through breast milk from mothers with Graves’ disease.4.The titer of thyroid autoantibodies in mothers’ breast milk would decrease withtime extending.5.The levels of thyroid autoantibodies in the mother with Graves’ disease couldaffect the offspring’s thyroid function through the placenta and breast milk.