CEBPA-Regulated Expression of Suppresses Milk Protein Synthesis Through MTOR and JAK2-STAT5 Signaling Pathways in Buffalo Mammary Epithelial Cells

Milk protein content is a key quality indicator of milk, and therefore elucidating its synthesis mechanism has been the focus of research in recent years. Suppressor of cytokine signaling 1 () is an important inhibitor of cytokine signaling pathways that can inhibit milk protein synthesis in mice. However, it remains elusive whether plays roles in the milk protein synthesis in the buffalo mammary gland. In this study, we found that the mRNA and protein expression levels of in buffalo mammary tissue during the dry-off period was significantly lower than those during lactation. Overexpression and knockdown experiments of showed that it influenced the expression and phosphorylation of multiple key factors in the mTOR and JAK2-STAT5 signaling pathways in buffalo mammary epithelial cells (BuMECs). Consistently, intracellular milk protein content was significantly decreased in cells with overexpression, while it increased significantly in the cells with knockdown. The CCAAT/enhancer binding protein α (CEBPA) could enhance the mRNA and protein expression of and its promoter activity in BuMECs, but this effect was eliminated when CEBPA and NF-κB binding sites were deleted. Therefore, CEBPA was determined to promote transcription via the CEBPA and NF-κB binding sites located in the promoter. Our data indicate that buffalo plays a significant role in affecting milk protein synthesis through the mTOR and JAK2-STAT5 signaling pathways, and its expression is directly regulated by CEBPA. These results improve our understanding of the regulation mechanism of buffalo milk protein synthesis.