Benzbromarone exacerbates high-fat-induced lipid accumulation in primary hepatocytes by activating PPARγ [OA]

Objective: Benzbromarone (BBR) is an effective uric acid-lowering drug, and studies have shown that BBR can exacerbate hepatic steatosis in obese individuals. However, the underlying mechanisms remain unclear. Methods: Primary hepatocytes isolated from C57/BL6 mice were cultured in vitro, and a high-fat culture model was simulated with 20mM oleic acid (OA) treatment. OA and bovine serum albumin (BSA) were mixed at a ratio of 1:4, pre-treated at 37°C for 1-2 hours, and then co-incubated with primary hepatocytes for 16 hours. The in vitro experiments were divided into four groups: control (BSA), control treatment group (BSA+BBR), high-fat group (OA), and high-fat treatment group (OA+BBR). After 16 hours of treatment, intracellular triglyceride (TAG) content was measured, and Bodipy staining was used to observe the number and morphological changes of lipid droplets. RNA-seq analysis was conducted to assess changes in the gene expression profile of hepatocytes with BBR treatment under BSA and OA conditions. Results: BBR significantly increased the intracellular TAG content in primary hepatocytes treated with OA, while no significant increase was observed in the BSA treatment group. Bodipy staining experiments also confirmed that BBR led to an increase in the number and size of lipid droplets under OA treatment. Transcriptomic data revealed that under both BSA and OA conditions, BBR upregulated the expression of a large number of genes related to lipid synthesis, which were associated with the activation of the PPARγ signaling pathway. Conclusion: BBR promotes the expression of genes involved in lipid synthesis in primary hepatocytes by activating PPARγ, leading to specific hepatic lipid accumulation under high-fat conditions.