Marchio Sophia P, El-Sheikh Ali Hossam, Scott Matthew A, Barbosa Fernandes Claudia, Scoggin Kirsten E, Troedsson Mats, Boakari Yatta
College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
Department of Veterinary Science, Maxwell H. Gluck Equine Research Center, University of Kentucky, Lexington, KY, USA.
Sci Rep. 2025 Aug 21;15(1):30714. doi: 10.1038/s41598-025-16671-5.
Despite its critical role in protecting the fetus, the amniotic membrane remains poorly understood in the context of disease response. The equine amniotic membrane is an important physical barrier to the amniotic compartment, and there is evidence that it may contribute to surfactant synthesis. Surfactants are essential for normal fetal lung development, and disruptions in its availability may be linked to future neonatal complications. Therefore, understanding the molecular changes that occur in fetal-maternal tissues during placentitis would clarify how this condition leads to abortion, preterm delivery, and stillbirth, and identify new strategies to manage the adverse outcomes. Thus, we used RNA sequencing, bioinformatic methods, and immunohistochemistry to characterize the equine amniotic membrane gene expression during experimentally induced ascending placentitis (placentitis group, n = 6) compared to gestationally matched control groups (control group, n = 6) at 288 days of gestation. We identified 288 differentially expressed genes (DEG) in the placentitis group compared to the control group. Placentitis was associated with the upregulation of toll-like receptors (TLR4), prostaglandin synthesis (PTGS2 and PTGES), apoptosis (MMP9 and CASP3), and hypoxia-associated genes (SOD2, BNIP3, and HMOX1). Our RNA sequencing results were supported by the visual identification of two of those proteins (TLR4 and PTGS2) in the immunohistochemistry analysis. Functional annotation revealed significant enrichment between the DEGs and the toll receptor signaling pathway, which may be a key factor negatively affecting placental functions. In conclusion, our study revealed that the amniotic membrane is not only a physical barrier but also plays an active role in immune response during ascending placentitis.
尽管羊膜在保护胎儿方面起着关键作用，但在疾病反应的背景下，人们对它的了解仍然很少。马的羊膜是羊膜腔的重要物理屏障，并且有证据表明它可能有助于表面活性剂的合成。表面活性剂对胎儿肺的正常发育至关重要，其可用性的破坏可能与未来的新生儿并发症有关。因此，了解胎盘炎期间胎儿-母体组织中发生的分子变化，将阐明这种情况如何导致流产、早产和死产，并确定管理不良后果的新策略。因此，我们使用RNA测序、生物信息学方法和免疫组织化学，来表征在妊娠288天时，与妊娠匹配的对照组（对照组，n = 6）相比，实验性诱导的上行性胎盘炎（胎盘炎组，n = 6）期间马羊膜的基因表达。与对照组相比，我们在胎盘炎组中鉴定出288个差异表达基因（DEG）。胎盘炎与Toll样受体（TLR4）、前列腺素合成（PTGS2和PTGES）、细胞凋亡（MMP9和CASP3）以及缺氧相关基因（SOD2、BNIP3和HMOX1）的上调有关。我们的RNA测序结果在免疫组织化学分析中对其中两种蛋白质（TLR4和PTGS2）的可视化鉴定得到了支持。功能注释显示，DEG与Toll受体信号通路之间存在显著富集，这可能是对胎盘功能产生负面影响的关键因素。总之，我们 的研究表明，羊膜不仅是一个物理屏障，而且在上行性胎盘炎期间的免疫反应中也发挥着积极作用。
