Regulation of Adipocyte Differentiation by Reversine in 3T3-L1 Cells

Reversine, a synthetic purine analog, is used for cell de-differentiation, and previous research has explored its effects on fat cell de-differentiation and trans-differentiation. However, limited studies have investigated the systematic control of fat cell differentiation during adipogenesis. In this study, we examined the impact of different concentrations and timings of reversine treatments on 3T3-L1 preadipocyte differentiation across various stages, including clonal expansion (day 0 - day 2), post-mitotic growth arrest (day 2 - day 4), and terminal differentiation phases (day 4 - day 6). We found that the differentiation of adipocytes depended on the concentration of reversine, with prolonged treatment leading to inhibited adipocyte differentiation. When treating specific stages with reversine, we observed the most significant decrease in intracellular lipid accumulation (p<0.05) and the expression of adipogenic marker genes (PPARγ, FASN, adiponectin) during the post-mitotic growth arrest phase (p<0.05). The impact of reversine treatment during the late differentiation phase was diminished compared to the two earlier stages of differentiation. However, high concentrations still exhibited inhibitory effects on intracellular lipid accumulation and adipogenic gene expression. Consistent reversine treatment from the early to late stages of differentiation effectively reduced the formation of adipocyte lipid droplets and associated gene expressions (p<0.05), depending on treatment duration and concentration. Our study suggests that reversine generally inhibits adipocyte differentiation, with maximal efficacy during the post-mitotic growth arrest phase.