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Obesity, a global health concern characterized by excessive fat accumulation, necessitates the discovery of anti-obesity com pounds. Rottlerin, known for its anti-cancer effects as a mitochondrial uncoupler, has been a subject of interest. However, its impact on reducing intracellular lipid accumulation remains a gap in our understanding. This study aimed to fill this gap by dissecting the mechanism of rottlerin in 3T3-L1 adipocytes. We treated differentiated 3T3-L1 cells with 0-20 mM of rottlerin for 48 hours to assess its capability to induce lipid accumulation. Notably, we observed no cytotoxicity associated with the treatment of rottlerin up to 20 mM, indicating its safety at these concentrations. Lipid accumulation, measured by oil Red O, was downregulated dose-independently by rottlerin. We also found that key lipogenic enzymes, including SCD1 and DGAT1, were decreased. The transcription factor of lipogenic genes, SREBP1, was reduced by approximately 80% with rottlerin. LRP6, a crucial link between de novo lipogenesis mechanism reactions and Wnt signaling, was also degraded by around 70%. Inter estingly, the downstream regulation of LRP6, b-catenin, and TCFL2 was diminished by rottlerin. Our data indicate that rottlerin alleviates adipocyte lipid accumulation by suppressing the LRP6/β-catenin/SREBP1c pathway. These findings underscore the potential of rottlerin as a safe nutraceutical for combating obesity.
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