Expression And Significance Of Galactose Agglutinin-3 In Endometrium Of Mice With Hashimoto's Thyroiditis

Objective Hashimoto's thyroiditis(HT)is an immune thyroid disease that is widely considered to affect the reproductive process of women.Galactose agglutinin-3(galectin-3)is a protein that regulates pregnancy.Therefore,the expression of galectin-3 in endometrium of HT patients may be affected by their disease.The purpose of this study was to investigate whether the expression of galectin-3 in endometrium of HT mice with normal thyroid function was affected by porcine thyroglobulin(pTg)in female NOD mice during pregnancy preparation.In order to explore the mechanism of HT affecting female reproduction to provide relevant experimental basis.Methods64 healthy NOD female mice of 8-9 weeks in age were randomly divided into two groups(by random number method): normal control group(CON group)and Hashimoto thyroiditis group(HT group),32 in each group.A mouse model of Hashimoto's thyroiditis was established by subcutaneous injection of porcine thyroglobulin(pTg)into the caudal root of mice.The modeling steps are as follows:25 ?g pTg is dissolved in100 ?l PBS solution,and the Freund's complete adjuvant(CFA)and pTg solution are mixed in the mortar at 1:1,then ground into "oil-in-water" form under the condition of ice bath,that is,complete emulsification.Then the emulsion was extracted with a syringe,and the mice were immobilized.0.1 ml emulsifier was injected subcutaneously into the tail of each mouse,1 cm away from the root of the tail.The control group was injected with the same amount of CFA.without pTg.On the 14 th day,the CFA of the model group was replaced with the Freund's incomplete adjuvant(IFA)to prepare theemulsion.The operation method was the same as before,and the control group was injected with the same amount of IFA,without pTg for 49 days.The estrus cycle of mice was determined by Rayleigh staining,the serum thyroid hormone T3,T4 level,antibody TgAb,TPOAb level and serum TSH level were measured by electrochemiluminescence immunoassay(ECLIA)and enzyme-linked immunosorbent assay(Elisa).The thyroid gland and endometrial gross specimen were prepared.The morphology of thyroid tissue and endometrium of mice were observed by HE staining.The expression of galectin-3 protein and mRNA in mouse endometrium were observed by immunohistochemical staining and reverse transcriptase polymerase chain reaction(RT-PCR).Flow microsphere capture technique(CBA)was used to determine the level of cytokines in uterine tissue.Result1.The results of Rayleigh staining in mouse endometrium during estrus cycle were as follows: preestrus: a large number of nucleated epithelial cells were seen under the microscope,and a large number of keratinized acellular epithelial cells were observed under the microscope,and in the estrus phase.Anestrus: nucleated epithelial cells,leukocytes,and acellular keratinocytes distributed evenly in the visual field;estrus interphase: on the basis of a large number of leukocytes infiltration,some nucleated epithelial cells were seen.2.Thyroid hormone and antibody levels in the two groups: compared with the normal control group,the TgAb,TPOAb level of the bridge group was increased(P < 0.05).3.Results of gross thyroid specimens and HE staining in the two groups: compared with the CON group,the thyroid tissue of the HT group was obviously enlarged,darker and more tenacious than that of the CON group.The results of HE staining showed that the infiltration of thyroid cells in HT group was more obvious than that in CON group,and the follicle of thyroid gland was damaged.4.Results of HE staining of endometrium in two groups: compared with the normal control group,the endometrium was thickened and the villus epithelium on both sides of the membrane could be connected,and the glands in the lamina propria were increased and larger.5.The distribution and expression of galectin-3 protein in endometrium of two groups:galectin-3 was expressed in each estrous cycle of mouse endometrium,and was brown granules,mainly distributed in the cytoplasm of luminal epithelium,glandular epithelium,especially apical surface.The cytoplasm of the interstitial cells was less distributed,but almost no distribution was found in the nuclei.Compared with the normal control group,the expression of galectin-3 protein decreased in estrus and estrus(P <0.05),but there was no significant difference in the expression of galectin-3 protein in estrus before estrus.6.The expression of galectin-3 mRNA in endometrium of the two groups: compared with the normal control group,the expression of galectin-3mRNA in the bridge group decreased during the four periods(P <0.05).7.Compared with the normal control group,the level of IL-2,IL-4 in the bridge group was lower than that in the control group(P <0.05).Conclusion The expression of galectin-3 in endometrium of mice with Hashimoto's thyroiditis was significantly increased during estrus,but decreased compared with the normal control group,which may be related to the changes of local cytokines in the uterus of Hashimoto thyroiditis mice.