This study investigates the inhibitory effects of polymethoxyflavones
(PMFs) on enzymes involved in the production of trimethylamine (TMA)
and trimethylamine-N-oxide (TMAO). PMFs were isolated from Valencia
orange peel and identified using column separation and NMR techniques.
The findings reveal that nobiletin and 3,6,7,8,2′,5′-hexamethoxyflavone
significantly suppress cntA/B and cutC/D, respectively. Furthermore,
3,6,7,8,2′,5′-hexamethoxyflavone decreases the level
of TMAO formation by suppressing the FMO3 mRNA level. This study elucidates
that specific structural features of PMFs can contribute to their
interactions with enzymes. Our study represents the first demonstration
of the ability of PMFs to mitigate the risk of cardiovascular disease
(CVD) by inhibiting enzymes responsible for TMA production, which
are generated by gut microbiomes. Furthermore, we introduce a novel
model system utilizing TMA-induced HepG2 cells to assess and compare
the inhibitory effects of PMFs on TMAO production. These findings
could pave the way for the development of novel therapeutic approaches
to manage CVD.