contrast, the mRNA expression levels of the SREBP-1c and ChREBP target genes, including acetyl-CoA carboxylase 1, fatty acid synthase, and stearoyl-CoA desaturase1, tended to decrease in the acetate-treated groups. The mRNA levels of ACC1 and FAS were significantly lower in the medium- and high-dose groups than in the control group and were significantly higher in the BML-275 and BML-275+acetate groups than in the control group. The mRNA levels of SCD-1 were significantly lower in the high-dose group and were markedly higher in the BML-275 group than in the control group. Collectively, these results SAR 405 site suggest that acetic acid inhibits the mRNA expression of lipid synthesis genes through inhibiting the expression and transcriptional activity of SREBP-1c and ChREBP in bovine hepatocytes. The phosphorylation level and enzyme activity of ACC1 in hepatocytes The phosphorylation level of ACC1 increased in an acetate dose-dependent manner and was significantly higher in the acetate-treated groups than in the control group. It was significantly lower in the BML-275 and BML-275+acetate groups than in the control group. ACC1 activity was significantly lower in the medium- and high-dose groups than in the control group and was markedly higher in the BML-275 and BML-275+acetate groups than in the control group. These results indicate that acetate-activated AMPKa can directly phosphorylate ACC1 and inhibit its activity, which inhibits lipid synthesis in bovine hepatocytes. Triglycerides content Acetate decreased hepatocyte TG content in a dose-dependent manner. The TG content was 35% lower in the high-dose acetate treatment group than in the control group. Taken together, our in vitro data demonstrate that acetic acid activates AMPKa signaling pathway to reduce TG content in bovine hepatocytes. Discussion In dairy cows, the liver is the main organ responsible for modulating lipid metabolism and maintaining lipid homeostasis Acetic Acid Activates the AMPK Signaling Pathway through responses to nutrient signals. In recent years, studies have shown 11784156 that acetic acid can act as a signaling molecule that modulates the expression of lipid metabolism genes in hepatocytes. An in vitro HepG2 cell study demonstrated that acetic acid activates AMPKa, which in turn upregulates the expression of lipid oxidation genes in the liver to reduce fat accumulation. A liver-specific AMPKa deletion in mice leads to increased plasma TG content and hepatic lipogenesis. The blood acetate concentration is dozens of times higher in dairy cows than that in humans and mice. Furthermore, the biological function of acetic acid in ruminants is different from that in humans and mice. However, it is not clear whether acetic acid activates the AMPK signaling pathway in the ruminant liver. In this study, we observed that the AMP/ATP ratio increased 2.00to 10.00-fold in acetate-treated hepatocytes. The phosphorylation level of AMPKa and AMPKa activity were 11078888 significantly increased in acetate-treated groups and were significantly lower in the BML275 and BML-275+acetate groups than in the control group. Abound acetate was converted to acetyl-CoA with the consumption of ATP in hepatocytes, resulting in a significant increase in the AMP/ATP ratio. 
The high AMP/ATP ratio increased AMPKa phosphorylation with the help of LKB1. These results demonstrate that acetic acid activates AMPKa in bovine hepatocytes. SIRT1 and AMPK is the cell metabolism regulator that regulates the cell energy metabolism.