2,4,6-tri-tert-butylphenol promoted adipogenesis in 3T3-L1 cells through a retinoid X receptor α-dependent mechanism

Mounting evidence suggests that certain environmental contaminants can influence adipogenesis and function as obesogens, many of which act by activating the key regulator peroxisome proliferator-activated receptor γ (PPARγ). Synthetic phenolic antioxidants (SPAs) are ubiquitously detected in both environmental matrices and human samples, whereas their potential effects on lipid metabolism and the underlying molecular mechanisms remain largely unexplored. In this study, we aimed to assess the agonistic effects of SPAs on PPARγ, glucocorticoid receptor (GR), and retinoid X receptor α (RXRα), as well as their adipogenic potential. Luciferase reporter gene assays showed that among the ten commonly used SPAs evaluated, only 2,4,6-tri-tert-butylphenol (AO 246) exhibited efficacious agonistic activity toward RXRα. Environmentally relevant level exposure to AO 246 significantly promoted adipogenesis in 3T3-L1 cells, as evidenced by increased lipid accumulation and adipogenic gene expression, which could be alleviated by the RXRα antagonist UVI 3003. Further analysis revealed that the early stage of differentiation (Days 0-2) was the critical window for AO 246 to enhance adipogenesis and modify the gene expression profile. This study first demonstrated the adipogenic effects of AO 246 by activating RXRα but not PPARγ or GR, emphasizing its possible implications in obesity and associated health risks.