Fyn在母胎界面的表达及免疫调控作用

The expression of Fyn at the fetomaternal interface and its roles in pregnancy immunity

目的研究Fyn在母胎界面的表达及作用。方法构建妊娠和自然流产小鼠模型,免疫组化、实时荧光定量PCR、western-blot检测子宫胎盘组织中Fyn表达;建立LPS诱导的小鼠流产模型,以Fyn活性抑制剂,计算胚胎吸收率,实时荧光定量PCR和流式细胞学技术检测炎症因子(IL-17、RORγt、IFN-γ和TNF-α)水平、Th17和Treg细胞含量。结果Fyn水平在小鼠妊娠第4.5天最高,着床后明显降低;Fyn主要表达于子宫系膜淋巴聚集区的淋巴细胞中,自然流产组Fyn表达高于正常妊娠组;SU6656抑制LPS诱导的小鼠流产和IL-17、RORγt、TNF-α的异常高表达,以及Th17细胞含量的升高。结论Fyn通过Th17细胞等免疫因素参与母胎免疫调控及妊娠维持。

Objective To observe Fyn expression at the fetomaternal interface and its potential roles.MethodsMurine models of normal pregnant and immunological abortion were established.Immunohistochemistry,real time quantitative PCR(qPCR)and western-blotting were applied to analyze Fyn expression in uteroplacental complexes;LPS was performed to induce immune imbalance and abortion of mice,selective Fyn inhibitor(SU6656)was injected simultaneously.The rates of fetal resorption were calculated,the mRNA levels of IL-17,RORγt,IFN-γand TNF-αwere examined through qPCR,and the percentages of Tregs and Th17 cells in CD4+lymphocyte of uteroplacental tissues were evaluated by flow cytometry.ResultsFyn level was highest among mice at E4.5,and then declined sharply after implantation,Fyn was expressed mainly by lymphocytes in mesometrial lymphoid aggregate of pregnancy.Compared to normal pregnant mice,the levels of Fyn were higher in abortive model mice;SU6656 rescued LPS-induced miscarriage and immunological imbalance at the fetomaternal interface.It inhibited the increased levels of cytokines(IL-17,RORγt and TNF-α)and proportion of Th17 cells induced by LPS.ConclusionFyn plays pivotal roles in pregnancy,through regulating maternal-fetal immunity,particularly Th17 cell differentiation and function.