Valine metabolite, 3-hydroxyisobutyrate, promotes lipid metabolism and cell proliferation in porcine mammary gland epithelial cells

Improving mammary gland epithelial cells proliferation through nutrition is an important approach for enhancing sow milk production and piglet growth. An intermediate metabolite of valine, 3-hydroxyisobutyrate (3-HIB), regulates cellular lipid metabolism. In the present study, we investigated the effects of 3-HIB on porcine mammary gland epithelial cells proliferation and lipid metabolism. The addition of an appropriate concentration of 3-HIB significantly increased mammary gland epithelial cell proliferation and the expression of proteins associated with cell proliferation. Compared to the control group, the addition of 0.4-0.8 mM 3-HIB increased the expression levels of mTOR signaling pathway-related proteins and the cell cycle protein, Cyclin D1, while inhibiting the expression of the cell cycle arrest protein, P27. The addition of 0.8 mM 3-HIB increased the triglyceride and lipid droplet content in the cells. The addition of 3-HIB increased the expression of proteins related to <i>de novo</i> fatty acid synthesis and transport, resulting in a marked increase in most polyunsaturated fatty acids in the 3-HIB-added group. Compared to the control group, the addition of 0.8 mM 3-HIB increased the expression levels of the fatty acid oxidation-related proteins, ACSL and CAD, ultimately increasing cellular ATP synthesis. In summary, the addition of 0.8 mM 3-HIB to porcine mammary gland epithelial cells promotes cell proliferation by enhancing lipid metabolism and the expression of cell proliferation-related proteins.