Liu Liping, Xu Hui
Department of Obstetrics, The Sixth Hospital of Wuhan, Affiliated Hospital of Jianghan University, Wuhan 430015, China.
Department of Obstetrics, Wuhan Children's Hospital (Wuhan Women's and Children's Health Care Center), Tongji Medical College, Huazhong University of Science & Technology, Wuhan 430015, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2025 Aug;41(8):724-734.
Objective To explore the alterations in macrophage polarization and the expression of decorin (DCN) protein in the decidua of patients with missed abortion (MA), as well as to elucidate the regulatory effect of DCN on macrophage polarization. Methods Flow cytometry was employed to assess the polarization ratio of decidual macrophages in MA, recurrent spontaneous abortion (RSA) and normal pregnancy (NP); The expression and localization of DCN and hypoxia-inducible factor 1α (HIF-1α) in decidua and villi were assessed by immunohistochemistry staining, while their protein levels were measured by Western blot. Primary trophoblasts and peripheral blood mononuclear cell (PBMC)-derived macrophages were isolated and cultured. ELISA was conducted to quantify DCN levels in the culture supernatant of primary trophoblast and PBMC-derived macrophages. Additionally, flow cytometry was applied to evaluate the polarization ratio of PBMC-derived macrophages. Immunofluorescence cytochemical staining was conducted to examine HIF-1α expression in macrophages. Western blot was performed to detect the expression of proteins related to the gene associated with retinoid-IFN-induced mortality 19 (GRIM-19)/signal transducer and activator of transcription 3 (STAT3)/HIF-1α signaling pathway in macrophages. Results The polarization ratio of M1 macrophages in the decidua of abortion patients was significantly higher than that of NP, whereas the ratio of M2 macrophages was significantly lower. The expression of DCN and HIF-1α protein were significantly evaluated in abortion patients compared to NP. The supernatant DCN content and HIF-1α protein expression of primary trophoblast and PBMC-derived macrophages cultured under 10 mL/L O for 24 hours were markedly increased compared to cells treated with 210 mL/L O. Compared with the PBS group, the proportion of M1 macrophage and GRIM-19 protein expression were significantly reduced in the DCN group, while phosphorylated STAT3 (p-STAT3) and HIF-1α protein expression were significantly increased. Conclusion The expression of DCN in decidua and villi of MA is higher than that of NP. DCN exhibits an inhibitory effect on the M1 polarization of PBMCs-derived macrophages, which is likely mediated through the GRIM-19/STAT3/HIF-1α signaling pathway.
目的 探讨稽留流产（MA）患者蜕膜中巨噬细胞极化及核心蛋白聚糖（DCN）蛋白表达的变化，并阐明DCN对巨噬细胞极化的调控作用。方法 采用流式细胞术评估MA、复发性自然流产（RSA）及正常妊娠（NP）患者蜕膜巨噬细胞的极化比例；通过免疫组织化学染色评估蜕膜和绒毛中DCN及缺氧诱导因子1α（HIF-1α）的表达及定位，同时采用蛋白质印迹法检测其蛋白水平。分离并培养原代滋养细胞和外周血单个核细胞（PBMC）来源的巨噬细胞。采用酶联免疫吸附测定（ELISA）法检测原代滋养细胞和PBMC来源巨噬细胞培养上清液中DCN水平。此外，采用流式细胞术评估PBMC来源巨噬细胞的极化比例。采用免疫荧光细胞化学染色检测巨噬细胞中HIF-1α表达。采用蛋白质印迹法检测巨噬细胞中与维甲酸-干扰素诱导死亡率19（GRIM-19）/信号转导子和转录激活子3（STAT3）/HIF-1α信号通路相关基因的蛋白表达。结果 流产患者蜕膜中M1巨噬细胞的极化比例显著高于NP患者，而M2巨噬细胞的比例显著低于NP患者。与NP患者相比，流产患者DCN和HIF-1α蛋白表达显著上调。与210 mL/L O₂处理的细胞相比，10 mL/L O₂处理24小时的原代滋养细胞和PBMC来源巨噬细胞的上清液DCN含量及HIF-1α蛋白表达明显增加。与磷酸盐缓冲液（PBS）组相比，DCN组M1巨噬细胞比例及GRIM-19蛋白表达显著降低，而磷酸化STAT3（p-STAT3）和HIF-1α蛋白表达显著增加。结论 MA患者蜕膜和绒毛中DCN的表达高于NP患者。DCN对PBMC来源巨噬细胞的M1极化具有抑制作用，其可能通过GRIM-19/STAT3/HIF-1α信号通路介导。
