miRNA-150_R-1 mediates the HIF-1/ErbB signaling pathway to regulate the adhesion of endometrial epithelial cells in cows experiencing retained placenta

Abstract

Retained placenta (RP) refers to reproductive disorders caused by the failure of fetal membranes to be expelled 12 h after delivery in dairy cows. Postpartum adhesion of the fetal membranes to the uterus causes diseases such as mastitis or endometritis, which threatening the profitability of the dairy industry. Emerging evidence suggests that micro RNAs (miRNAs) play crucial roles in various processes, such as the occurrence and progression of fetal membranes discharge. However, the molecular mechanisms of miRNAs in RP remain unknown. In this study, we performed RNA-sequencing to characterize the expression profiles of mRNAs and miRNAs in caudal vein blood samples of postpartum Holstein cows whose fetal membranes were discharged normally or retained to identify RP-related genes and evaluate their molecular mechanisms. We identified 44 differentially expressed miRNAs (19 upregulated and 25 downregulated) and 706 differentially expressed mRNAs (325 upregulated and 381 downregulated) in the RP group compared to the normal fetal membranes discharge group. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis revealed that differentially expressed mRNAs were mainly enriched in the extracellular matrix, cell adhesion, and autoimmunity-related biological processes or pathways. Further analyses using RNA-sequencing, a dual luciferase reporter system, quantitative reverse transcription-PCR, immunofluorescence, and western blotting verified that endothelial PAS domain protein 1 (EPAS1) is regulated by miR-150_R-1 in endometrial epithelial cells. We demonstrated the relationship between EPAS1 and RP and confirmed that EPAS1 is upregulated in the blood and placenta of cows that experience RP. Further, we proposed a model of the miRNA-mRNA negative regulatory network mediated by the HIF-1/ErbB signaling pathway to show its regulatory role in RP.

circAMN1-Mediated Ferroptosis Regulates the Expulsion of Placenta in Trophoblast Cells

“…Twelve DE circRNAs were also randomly selected for qPCR validation, and the circRNA expression was consistent with the RNA-Seq data trend (Figure 1A-D and Table S4). Using previous research [1], the datasets of GSE214588 were downloaded from the Gene Expression Omnibus (GEO) (https://www.ncbi.nlm.nih.gov/gds/?term=, accessed on 18 October 2022), RNA-Seq data, and the biological characteristics of PCD were used to identify DE genes in these datasets. We focused on the involvement of DE miRNA target genes in apoptosis, cell differentiation, multicellular development, the positive regulation of cell surface and cell population proliferation, the negative regulation of cell apoptosis, chromatin, and the membrane (Figure 1E).…”

“…The separation between the maternal and fetal placentas after fetal delivery is a prerequisite for the smooth expulsion of the placenta [1,2]. During the expulsion of the placenta, the ability to induce the proliferation, invasion, and migration of trophoblast and endometrial epithelial cells is weakened, increasing cell death and causing the fetal cotyledon to detach from the maternal uterine caruncle, thereby promoting placental expulsion [3].…”

“…In this study, we aimed to identify candidate differentially expressed (DE) genes related to PCD in cows with normal fetal membrane discharge (NC) or RP based on previous research [1], RNA sequencing (RNA-Seq), GSE214588 dataset, and bioinformatic analysis. Furthermore, we aimed to elucidate the mechanisms associated with ferroptosis in trophoblast cells in vitro.…”

Hypoxia and the endometrium: An indispensable role for HIF-1α as therapeutic strategies

“…In patients with chronic endometritis, an increase in peri-implantation endometrial HIF-1α has been observed, which is a crucial trigger for the upregulation of VEGF, resulting in excessive endometrial vascularization and infertility [ 81 ]. The activation of the HIF-1 pathway also upregulates epidermal growth factor receptor and focal adhesion kinase, resulting in endometrial epithelial cells adhering to trophoblast cells and a retained placenta, triggering an inflammatory response [ 82 ]. In addition, in animal models with endometritis depending on the administration of lipopolysaccharide, Jiang et al [ 83 ] suggested a molecular mechanism involved in upregulating HIF-1α levels.…”

Editorial: Reproduction-related non-coding RNAs in animals

“…The sequencing result demonstrated the relationship between EPAS1 and retained placenta, and confirmed that EPAS1 was upregulated in the blood of cattle experiencing retained placenta. In addition, they found that EPAS1 was negatively regulated by miR-150_R-1, and then regulated the changes of related molecules in HIF-1/ErbB signaling pathway, especially the abnormal changes of EGFR and p-FAK, which affected the adhesion of fetal placental cotyledon to uterine nodules after delivery, leading to retained placenta ( 1 ).…”