Sulforaphane and myricetin act synergistically to induce apoptosis in 3T3-L1 adipocytes

The aim of the present study was to investigate whether sulforaphane (SFN) and myricetin (Myr) synergistically induce apoptosis in adipocytes. The viability of mature 3T3-L1 adipocytes treated with 40 µM SFN and/or 100 µM Myr was assessed using an MTT assay. Apoptosis was assessed by Hoechst 33258 nuclear staining, and by detection of single-stranded DNA using an enzyme-linked immunosorbent assay. Compared with the effects of each compound alone, the combination of SFN and Myr synergistically reduced cell viability, induced apoptosis, increased pro-apoptotic Bcl-2 associated X protein expression, decreased anti-apoptotic B-cell lymphoma-2 expression, enhanced Bcl-2-associated death promoter (Bad) translocation from the cytoplasm to the mitochondria, and reduced Bad phosphorylation at Ser112. These effects were accompanied by increased cleavage of caspase 3 and poly-ADP-ribose-polymerase. In addition, combined SFN and Myr treatment significantly decreased the protein expression levels of phosphorylated AKT serine/threonine kinase 1 (Akt) at Ser473, as well as the phosphorylation of the downstream protein ribosomal protein, S6 kinase β-1. Therefore, SFN plus Myr was a more potent inducer of apoptosis in 3T3-L1 adipocytes than either compound alone. The results of the present study suggest that the mechanism of SNF/Myr-induced apoptosis involved activation of the Akt-mediated mitochondrial apoptotic pathway. This may aid treatment of animal models of obesity and preclinical testing.