不完全型胚胎着床障碍模型小鼠滤泡辅助性T细胞占比及功能相关细胞因子表达水平分析OA

Objective To detect the proportion of splenic follicular helper T(Tfh)cells and their functionally re-lated cytokine expression levels in the incomplete embryo implantation disorder(EID)model mice,and to explore the immunological mechanism of Tfh in infertility caused by embryo implantation disorder.Methods Sixteen fe-male Kunming mice were randomly divided into two groups,with eight mice in each group.On day 4 of pregnancy,an incomplete EID mouse model was established by oral gavage of mifepristone suspension,while an equal volume of saline was administered to the control group.On day 8 of pregnancy,the mice were euthanized.Flow cytometry was used to detect the levels of Tfh cells in the spleen lymphocytes of both incomplete EID mice and normal control mice.qRT-PCR was performed to measure the mRNA levels of B-cell lymphoma 6(Bcl-6)and C-X-C chemokine receptor type 5 protein(CXCR5)in the spleen lymphocytes of both groups.Western blot was employed to assess the protein expression levels of Bcl-6 and CXCR5 in the spleen lymphocytes of both groups.Serum levels of inter-leukin-4(IL-4),IL-6,and IL-21 were measured by ELISA.Immunohistochemistry(IHC)was used to observe the expression levels of progesterone receptor(PR),Bcl-6,and CXCR5 proteins in the uterine endometrial tissue of mice in both groups.Results Incomplete-type EID mice had a reduced number of embryo implantation points and reduced endometrial PR expression.Flow assay results showed that the proportion of CD4+CXCR5+Tfh cells in splenic lymphocytes of incomplete-type EID mice was significantly higher than that of normal controls(P<0.05).Compared with the normal control group,Bcl-6 and CXCR5 mRNA levels and protein levels were elevated in splenic lymphocytes of incomplete EID mice,with statistically significant differences(P<0.05);serum IL-4,IL-6,and IL-21 levels were elevated in incomplete EID mice,and Bcl-6 and CXCR5 proteins in the endometrium were significantly elevated(P<0.05).Conclusion The increase of Tfh cells and their associated cytokines Bcl-6 and CXCR5 is associated with the development of incomplete EID,and may be involved in the development of fe-male immune infertility.