Effect of acetate, β-hydroxybutyrate and their interaction on lipogenic gene expression, triglyceride contents and lipid droplet formation in dairy cow mammary epithelial cells

Abstract

The purpose of this study was to assess the effects of acetate and β-hydroxybutyrate alone or in combination on lipogenic genes and their associated regulatory proteins in dairy cow mammary epithelial cells (DCMEC) using quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blotting, lipid droplet staining and a triglyceride content detection kit, to determine whether SCFA are related to milk fat synthesis regulation in DCMEC. Our experiment shows that addition of different concentrations of acetate, β-hydroxybutyrate and their combinations to DCMEC increase in relative mRNA abundance of lipogenic genes and key transcription factors suggest an increase in lipogenic capacity, which is supported by an increased in cytosolic triglyceride content. Similarly, the protein expression level of acetyl-coenzyme A carboxylase (ACACA), fatty acid synthase (FASN) and sterol-coenzyme desaturase-1 (SCD1) genes and the transcription factor sterol regulatory element-binding protein-1 (SREBP1) were found to be increased by addition of acetate, β-hydroxybutyrate and their combinations. The expression pattern of fat-related genes and proteins showed similar trends in almost all treatments, suggesting that common transcription factor are regulating these genes. These results show that acetate and β-hydroxybutyrate regulate fat synthesis, further confirming that SCFAs work by targeting genes to activate the SREBP1 and insulin-induced gene 1 protein (INSIG1) signalling pathways in DCMEC.