Study on the anti-inflammatory effects and mechanisms of gentisic acid based on the LPS-induced RAW264.7 cell inflammation model and the oxazolone-induced zebrafish inflammation model.

BACKGROUND

Gentisic acid (GA), a natural polyphenolic compound, possesses significant anti-inflammatory activity, but its molecular mechanism, particularly its association with the intestinal microbiota and specific signaling pathways, remains unclear.

PURPOSE: In this study, we aimed to clarify anti-inflammatory targets, pathways and mechanism of GA using network pharmacology, in vitro and in vivo experiments, and microbiome analysis.

MATERIALS AND

METHODS

Network pharmacology was used to predict the anti-inflammatory targets and pathways of GA. Lipopolysaccharide (LPS)-induced RAW 264.7 cell models and oxazolone (OXA)-induced zebrafish models were established to evaluate the anti-inflammatory effects of GA. Quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting, and 16S rDNA sequencing were used to detect changes in key genes, proteins, and intestinal microbiota, respectively.

RESULTS AND DISCUSSION: GA significantly attenuated LPS- and OXA-induced inflammation, reducing nitric oxide (NO) levels and inflammatory cytokines (IL-1β, TNF-α, and IL-6) in RAW 264.7 cells, along with IL-6 and IL-1β levels in zebrafish intestinal tissues. It was associated with the regulation of the IL-17 signaling pathway, reducing the abundance of Acinetobacter, Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, and Shewanella, which were positively correlated with key genes (IKKα, IKKβ, TRAF6, etc.) in the IL-17 pathway.

CONCLUSION

GA may exert anti-inflammatory effects potentially via correlatively regulating the composition of pathogenic intestinal bacteria to maintain microecological homeostasis and modulating the IL-17/NF-κB pathway. These correlative findings provide a valuable reference for the research and development of polyphenolic compounds.