| Objective Hashimoto’s thyroiditis(HT)is an autoimmune thyroid disease(AITD)characterized by the presence of high titers of autothyroid antibodies(TPOAb,Tg Ab)in the serum.HT often shows abnormal mood,especially anxiety and depression.The level of serum TPOAb antibody in pregnant women was significantly related to emotional disorders.The frontal lobe is a key brain area that affects emotional behavior.Glutamic acid(Glu)is an important neurotransmitter in the central nervous system,which plays an important role in the pathogenesis of anxiety and depression.Glu mainly depends on Glu transporter to regulate and maintain the low concentration of extracellular nerve cells.Therefore,the purpose of this study is to establish a pregnant HT animal model,observe the changes of Glu and its transporter in frontal lobe,and try to explore the effects of HT on Glu and its transporter in frontal lobe in pregnant mice.Methods After one week of adaptive feeding,50 clean grade healthy nod female mice(6 weeks old)were randomly divided into two groups,namely 20 normal control group(CON group)and 30 Hashimoto thyroiditis group(HT group).Porcine thyroglobulin(p Tg)was injected subcutaneously in the tail root of HT group mice to prepare HT mouse model.The specific operation steps are as follows:Dissolve p Tg reagent in normal saline(100 μl normal saline dissolves 25 μg p Tg powder),inhale the same amount of Freund’s complete adjuvant(CFA)and p Tg solution into a disposable syringe,connect the three-way tube,ice bath Grind until completely emulsified under the conditions,that is,the white emulsion is dropped into cold water and can float on the surface of the liquid for a long time without spreading.The mice in the HT group were fixed,and 0.1 ml of emulsion was injected subcutaneously with a disposable syringe 1 cm from the base of the tail of the mouse.The same amount of CFA emulsifier without p Tg was injected into the CON group mice at 1 cm from the base of the tail.After 14 days,replace CFA with Incomplete Freund’s Adjuvant(IFA)and prepare emulsifier with p Tg.In the same way,inject 0.1 ml of emulsifier subcutaneously at 1 cm in the tail root of each HT group mouse and inject in CON group Equal amount of IFA emulsifier without p Tg.The modeling period is 49 days.After successful modeling,the female rats of each group and the healthy NOD male rats were caged in a 2: 1ratio,and the pregnancy model was determined by observing the Yin plug.Serum thyroid stimulating hormone(TSH)levels in pregnant mice were detected by enzyme-linked immunosorbent assay;serum thyroid hormones triiodothyronine(T3),thyroxine(T4)and thyroid peroxidase were measured by electrochemiluminescence immunoassay Antibody(TPOAb)and thyroglobulin antibody(Tg Ab)levels;preparation of gross thyroid specimens,observation of thyroid tissue morphology in mice under HE staining method;detection of Glu concentration in frontal lobe of pregnant mice by enzyme-linked immunosorbent assay;detection by Western blot The protein expression levels of vesicle glutamate transporter VGLUT 1 and excitatory glutamate transporter EAAT 2 in the frontal lobe of pregnant mice;RT-q PCR was used to observe the expression of VGLUT 1 and EAAT 2 m RNA levels in the frontal lobe of pregnant rats.Result1.Thyroid antibody and hormone level of pregnant rats in two groups: serum thyroid antibody level(TGAb,TPOAb)in HT group was higher than that in con group(P <0.05),and there was no significant difference in serum thyroid hormone(T3,T4,TSH)between the two groups;2.Compared with con group,the level of glutamate in frontal lobe of HT group increased(P < 0.05);3.Description and he staining results of thyroid samples of two groups of pregnant rats: compared with con group,the thyroid samples of HT group were darker in color,larger in volume and stronger in texture;he staining results of thyroid: in HT group,part of thyroid follicles were damaged and a large number of lymph cells were infiltrated in thyroid,and the thyroid follicles of con group were relatively complete.4.The expression of glutamate transporter protein in frontal lobe of two pregnant rats:the expression level of VGlu T1 protein in frontal lobe of HT group was lower than that of con group(P < 0.05),and there was no significant difference in the expression level of EAAT2 protein in frontal lobe of two pregnant rats.5.The m RNA expression of the frontal lobe glutamate transporter in the 2 groups of pregnant rats: Compared with the CON group,the m RNA expression of VGLUT 1 in the frontal lobe of the pregnant rats in the HT group was reduced(P < 0.05).There was no significant difference in the expression level of EAAT 2 m RNA in the frontal lobe of the two groups of pregnant rats.Conclusion HT can increase the Glu concentration in the frontal lobe of pregnant rats,and the protein expression of the glutamate transporter VGLUT 1 decreases,suggesting that HT can cause Glu metabolism in the frontal lobe. |
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