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Dencichine attenuates the virulence of Fusobacterium nucleatum by targeting hydrogen sulfide-producing enzyme

Wang Minyu,Chu Weihua

Oral opportunistic pathogen Fusobacterium nucleatum can participate in various disease processes through the metabolite hydrogen sulfide, such as halitosis and colorectal cancer. The object of this study is to identify inhibitor capable of suppressing Fn1220, which is the principal hydrogen sulfide-producing enzyme in F . nucleatum . Through this inhibition, we aim to reduce the hydrogen sulfide production of F . nucleatum , consequently diminishing its virulence. Employing molecular docking techniques for inhibitor screening, we identified dencichine as the monomeric compound from Chinese medicine exhibiting the lowest binding energy to Fn1220 among a set of 27,045 candidates, and evaluated in vitro the ability of dencichine to inhibit hydrogen sulfide production using bismuth chloride method. Additionally, we investigated its impact on key virulence factors, including biofilm formation, hemolysis, and adhesion factors of F . nucleatum , using the crystalline violet method, sheep blood method, and RT-qPCR, respectively. Furthermore, we assessed the influence of dencichine on the lifespan of Caenorhabditis elegans . Results showed that dencichine was a suitable inhibitor of the Fn1220 of F . nucleatum , which significantly inhibited the production of virulence factors, e.g., biofilm, hemolysin, FadA, and Fap2 of F . nucleatum and improved the survival of C . elegans . We successfully identified the inhibitor of the enzyme Fn1220, dencichine, which inhibited the production of hydrogen sulfide and attenuated the virulence of F . nucleatum and holds promise as a potential therapeutic avenue for addressing oral diseases, e.g., halitosis in the future.