Potential effects on the signaling network mediated by overexpression of the vitronectin gene in Hu sheep ruminal epithelial cells using multi-omics analysis

Vitronectin (VTN) is a multifunctional extracellular matrix protein involved in cell adhesion, migration, and signal transduction. In this study, we constructed and transfected a <i>VTN</i> overexpression vector in Hu sheep ruminal epithelial cells (RECs) and performed a multi-omics analysis integrating transcriptomics and metabolomics. Compared with controls, 495 differentially expressed genes (DEGs) were identified (241 upregulated and 254 downregulated), primarily enriched in adhesion/mechanotransduction pathways such as ECM-receptor interaction and focal adhesion, as well as amino acid transport and membrane-related complexes; in contrast, translational preparatory processes including the spliceosome and aminoacyl-tRNA biosynthesis were suppressed. Metabolomics identified 103 differential metabolites (53 upregulated and 50 downregulated), prominently involving glycerophospholipid metabolism, nucleotide sugar biosynthesis, GPI-anchor biosynthesis, autophagy, and retrograde endocannabinoid signaling, indicating reinforced membrane lipid remodeling and membrane protein targeting. Multi-omics integration indicates that <i>VTN</i>, by remodeling ECM and membrane lipids, is associated with enhanced integrin-focal adhesion signaling and mechanotransduction, optimizes mitochondrial ATP production and energy utilization, and directs a programmed reconfiguration of lipid metabolism; concurrently, endocannabinoid-related pathways and "neurotransmission-like" signals such as NA-GABA were upregulated, providing an inhibitory/buffering tone against inflammation and environmental stress. Overall, <i>VTN</i> establishes a multilayered "adhesion-metabolism-repair" regulatory network that promotes rapid renewal and injury repair of RECs, offering a mechanistic basis and potential molecular targets for enhancing rumen function and production performance in ruminants.