Wang Qingchen, Feng Weimin, Tan Yuan, Qiao Jiao, Liu Hongchao, Liu Qi, Wang He, Zhang Qian, Tao Jingjin, Li Zhongxin, Yang Boxin, Xu Zhen, Wang Chong, Yang Shuo, Cui Liyan
Department of Laboratory Medicine, Peking University Third Hospital, Beijing, 100191, China.
Institute of Medical Technology, Peking University Health Science Center, Beijing, 100191, China.
J Transl Med. 2025 May 23;23(1):581. doi: 10.1186/s12967-025-06596-y.
Antiphospholipid syndrome (APS) is an autoimmune disease primarily manifested by recurrent thrombosis and pregnancy-related complications. The migration and invasion abilities of trophoblast cells play a crucial role in maintaining normal pregnancy. It is now increasingly recognized that adverse pregnancy outcomes in APS are associated with the disruption of trophoblast function by aPL (antiphospholipid antibodies), rather than thrombotic occlusion of the placental vasculature. Therefore, this article aimed to explore the potential mechanisms by which aPL affect trophoblast cell function.
An APS cell model was established in the HTR-8 trophoblast cell line, followed by RNA sequencing to identify key genes involved in trophoblast cell function. To explore the underlying mechanisms, we employed quantitative real-time PCR, Western blotting, immunohistochemistry, ELISA, plasmid transfection and KEGG pathway enrichment analysis. Functional assays, including migration and invasion tests, were conducted to evaluate trophoblast cell ability. Clinical samples were collected, and the expression levels of target molecules in serum were quantified using ELISA. Additionally, an APS animal pregnancy model was developed to assess pregnancy loss rates and analyze the expression of specific target genes in the placenta.
Sequencing analysis revealed significant downregulation of MMP1 in the APS model, confirmed by qPCR and Western blotting. Correspondingly, migration and invasion of HTR-8 cells were impaired in the APS group, but MMP1 overexpression restored trophoblast cell function. Serum MMP1 levels were lower in APS patients than in controls. In the animal pregnancy model, the APS group exhibited higher pregnancy loss, with placental immunohistochemistry confirming decreased MMP1 expression. KEGG enrichment analysis of differentially expressed genes between the NC and APS groups revealed a significant difference in the MAPK pathway, with P-JNK showing the most notable reduction. C-Jun, a downstream regulator of JNK, also decreased and modulated MMP1 expression. Notably, Anisomycin treatment increased P-C-Jun, upregulated MMP1, and enhanced trophoblast migration and invasion.
APL downregulated MMP1 expression by suppressing the JNK/C-Jun signaling pathway in trophoblast cells, thereby reducing their migratory and invasive capabilities. This represent a potential pathogenic mechanism contributing to adverse pregnancy outcomes in APS patients, highlighting possible therapeutic targets for intervention in APS management.
抗磷脂综合征（APS）是一种自身免疫性疾病，主要表现为反复血栓形成和与妊娠相关的并发症。滋养层细胞的迁移和侵袭能力在维持正常妊娠中起着关键作用。现在越来越认识到，APS患者不良妊娠结局与抗磷脂抗体（aPL）破坏滋养层细胞功能有关，而非胎盘血管系统的血栓闭塞。因此，本文旨在探讨aPL影响滋养层细胞功能的潜在机制。
在HTR-8滋养层细胞系中建立APS细胞模型，随后进行RNA测序以鉴定参与滋养层细胞功能的关键基因。为探究潜在机制，我们采用了定量实时PCR、蛋白质免疫印迹法、免疫组织化学、酶联免疫吸附测定、质粒转染和KEGG通路富集分析。进行了包括迁移和侵袭试验在内的功能测定，以评估滋养层细胞能力。收集临床样本，并使用酶联免疫吸附测定法定量血清中靶分子的表达水平。此外，建立了APS动物妊娠模型以评估妊娠丢失率，并分析胎盘中特定靶基因的表达。
测序分析显示APS模型中MMP1显著下调，定量PCR和蛋白质免疫印迹法证实了这一点。相应地，APS组中HTR-8细胞的迁移和侵袭受损，但MMP1过表达恢复了滋养层细胞功能。APS患者血清MMP1水平低于对照组。在动物妊娠模型中，APS组妊娠丢失率更高，胎盘免疫组织化学证实MMP1表达降低。对NC组和APS组之间差异表达基因的KEGG富集分析显示，丝裂原活化蛋白激酶（MAPK）通路存在显著差异，磷酸化JNK（P-JNK）下降最为显著。JNK的下游调节因子C-Jun也减少并调节MMP1表达。值得注意的是，茴香霉素处理增加了磷酸化C-Jun（P-C-Jun），上调了MMP1，并增强了滋养层细胞的迁移和侵袭。
aPL通过抑制滋养层细胞中的JNK/C-Jun信号通路下调MMP1表达，从而降低其迁移和侵袭能力。这代表了导致APS患者不良妊娠结局的一种潜在致病机制，突出了APS治疗中可能的干预治疗靶点。
