母-胎界面GAS6低表达诱导蜕膜巨噬细胞分化异常致自然流产的机制研究

Downregulated Expression of GAS6 in Maternal-Fetal Interface Affects the Polarization of Decidual Macrophagesand Causes Natural Abortion

目的:探讨早孕期母-胎界面中蜕膜基质细胞(DSCs)的生长停滞特异性基因产物6(GAS6)表达对蜕膜巨噬细胞(dMφ)分化的调控作用。方法:收集分泌期子宫内膜和正常早孕妇女蜕膜,运用免疫组化观察GAS6的定位及表达情况。再收集早期自然流产(ESA)患者和正常早孕妇女的蜕膜组织,运用免疫组化、定量聚合酶链反应(qPCR)、Westen bolt比较DSCs中GAS6的表达情况;同时运用qPCR比较dMφ上GAS6受体AXL、MERTK和TYRO3的表达情况。随后用不同浓度重组人GAS6蛋白(rhGAS6)刺激dMφ,多色流式细胞技术分析巨噬细胞的表型变化。结果:与分泌期子宫内膜间质细胞相比,正常早孕期母-胎界面蜕膜基质细胞GAS6的表达水平显著升高。与正常早孕期相比,ESA患者DSCs中GAS6表达水平显著降低;与此同时,ESA患者dMφ上GAS6的受体MERTK显著降低,AXL、TYRO3也降低,但差异无统计学意义。rhGAS6处理dMφ后,细胞表面分子CD163的表达水平显著上升,抑炎细胞因子TGF-β_(1)的分泌水平显著上升。结论:正常妊娠早孕期DSCs分泌的GAS6可促进巨噬细胞向M2型分化,参与免疫耐受的建立与妊娠的维持。母-胎界面GAS6的异常低表达可能参与了ESA的发生。

Objective:To explore the expression of Growth Arrest-Specific 6(GAS6)in Decidual Stromal Cells(DSCs)in early pregnancy and how GAS6 affects the function of Decidual macrophages(dMφ).Methods:The secretory endometrium and the decidua of early pregnant were collected,the localization and expression of GAS6 were observed by immunohistochemistry(IHC).Then the deciduaofpatients with early spontaneous abortion(ESA)and controlgroupwere collected to compare the expression of GAS6 in DSCs by IHC,q-PCR and Western boltting.At the same time,q-PCR was used to compare the expression of GAS6 receptors(AXL,MERTK and TYRO3)in dMφ.Subsequently,dMφs were stimulated with different concentrations of recombinant human GAS6 protein(rhGAS6),and the phenotypic changes of macrophages were analyzed by multicolor flow cytometry.Results:Comparewith endometrial stromal cells in the secretory stage,the expression of GAS6 of DSCs in early pregnancy was significantly increased.Compared with normal early pregnancy,the expression of GAS6 in DSCs with ESA was significantly decreased.MERTK,one of receptors of GAS6 on dMφ,was significantly reduced in ESA.The expression of AXL and TYRO3 were also reduced,while the statistics had no significant change.Multicolor flow cytometry technology showed that CD163,TGF-β1 considerably increased after stimulated by rhGAS6.Conclusions:GAS6 secreted by DSCs in early pregnancy can promote dMφinto M2.GAS6 can participate in the establishment of immune tolerance and maintenance of pregnancy.In addition,the low expression of GAS6 in the materal-fetal interface may be involved in the occurrence of ESA.