We previously demonstrated that vitamin B6 suppresses tumorigenesis in the colon of mice and exerts an anti-inflammatory effect through the inhibition of NF-κB activation. As these effects resemble the pharmacological properties of thiazolidinedione (TZD), a synthetic peroxisome proliferator-activated receptor-γ (PPARγ) ligand, this study was designed to examine the effect of vitamin B6 on the activation of PPARγ and adipogenesis in 3T3-L1 adipocyte cells. Pyridoxal 5'-phosphate (PLP), one of the vitamin B6 derivatives, was shown to promote adipogenesis in the 3T3-L1 adipocytes. In addition, PLP specifically induced mRNA expression of PPARγ target genes in the 3T3-L1 adipocytes and enhanced the lipid accumulation and adipocyte fatty acid-binding protein (aP2) mRNA expression in NIH3T3 cells stably expressing PPARγ. Furthermore, the administration of vitamin B6 increased the expression of aP2 mRNA in mouse adipose tissues. Collectively, these observations suggest a novel function of vitamin B6 as an activator for PPARγ, which may contribute to the anti-tumor and anti-inflammatory effects of vitamin B6.