Polydatin Targets <scp>ACADVL</scp> to Combat Obesity by Promoting Adipocyte Browning and Activating Fatty Acid Oxidation

Stimulating white adipose tissue (WAT) browning represents a promising therapeutic strategy for treating obesity and related disorders. Polydatin (PD), a natural bioactive compound, exhibits anti-inflammatory and lipid-modulating properties; however, its impact on adipocyte browning and the underlying mechanisms remain unclear. In this study, we aimed to investigate the effect and mechanism of PD on diet induced obesity (DIO) with focus on adipocyte browning. In DIO mice, the effect of PD on energy expenditure and thermogenesis was measured using metabolic monitoring instruments and cold exposure test. The morphology of adipose tissue was observed by H&E staining and transmission electron microscopy. The effect of PD on C3H10T1/2 MSCs-differentiated adipocytes was verified by ORO and Mito Tracker staining, and OCR measurement. Target identification was performed by limited proteolysis-mass spectrometry (LiP-MS). The content of fatty acids was determined using lipidomics. Gene knockdown was achieved via lentiviral transduction. PD alleviated metabolic impairment and enhanced energy expenditure via promoting inguinal WAT (iWAT) browning in DIO mice. This process, driven by coordinately upregulating UCP1 and PGC1α, enhancing mitochondrial function, and increasing energy expenditure, was critically depended on fatty acid oxidation (FAO) activation. Using Lip-MS, molecular docking, cellular thermal shift assay (CETSA) and Bio-layer interferometry (BLI), acyl-CoA dehydrogenase very long-chain (ACADVL) was identified as a direct target of PD. Crucially, Acadvl knockdown abrogated PD-driven browning and FAO activation. Activation of ACADVL via natural compounds, PD for instance, could provide promising and inexpensive strategies in the treatment of obesity and metabolic disorders.