α-Naphthoflavone modulates inflammatory response in adipocytes-macrophages interaction through NFκB signaling

Our previous study demonstrated that α-naphthoflavone (α-NF) inhibits mouse 3T3-L1 pre-adipocytes differentiation via PPARγ, a key transcription factor in adipogenesis. Due to the critical role of inflammation in adipogenesis, we speculated that the suppression role of α-NF in adipogenesis might involve in modulation of cytokines secretion raised by adipocyte differentiation cocktail. Therefore, the present study aims to investigate the role of α-NF in modulating of inflammatory response during adipocytes differentiation and adipocyte-macrophage interaction.

Conditioned medium from different doses of α-NF treated 10-day differentiated 3T3-L1 adipocytes were collected to culture RAW264.7 macrophages. Conditioned medium from activated macrophages and α-NF pre-treated macrophage were used to investigate the effects of α-NF in adipocytes differentiation. Cultured cells and medium were harvested for RT-PCR, Western blot and ELISA.

α-NF dose-dependently decreased TNF-α and IL-6 and increased IL-10 expression induced by IDM (Insulin, dexamethasone, isobutylmethylxanthine) in 3T3-L1 pre-adipocytes. Conditioned medium from α-NF treated 3T3-L1 differentiated cells inhibited inflammatory response in mouse macrophage cell line RAW264.7 in contrast to IDM control medium. NFĸB activation elicited by IDM was suppressed by α-NF in a dose-response manner. Consequently, decreased TNF-α and increased IL-10 secretion, downstream targets of NFĸB signaling pathway, were observed with α-NF in macrophages. Finally, Conditioned medium from α-NF pre-treated, LPS-activated macrophages ameliorated the suppression of 3T3-L1 adipogenesis by LPS activated macrophages.

Our results suggest that α-NF regulates inflammation response in both adipocytes and macrophages and adipocyte-macrophage interaction which contributes to pre-adipocyte differentiation.