GEN‐27 exhibits anti‐inflammatory effects by suppressing the activation of NLRP3 inflammasome and NF‐κB pathway

Abstract Prolonged inflammation and deregulated cytokine production are associated with diversified inflammatory diseases. Genistein (GEN), the active and predominant isoflavonoid in dietary soybean, possesses anti‐inflammatory activity. Our study aimed to assess the anti‐inflammatory effects of GEN‐27, a derivative of GEN, as well as explore the potential molecular mechanisms using lipopolysaccharide (LPS)‐induced RAW264.7 cells. In our study, we demonstrated that GEN‐27 administration (1, 5, or 10 μM) dose‐dependently inhibited nitrite and nitric oxide (NO) levels in LPS‐stimulated RAW264.7 cells. Also, GEN‐27 suppressed the release of LPS‐induced pro‐inflammatory cytokines including tumor necrosis factor‐α (TNF‐α), interleukin‐1β (IL‐1β), IL‐6, and IL‐18. Moreover, GEN‐27 attenuated LPS‐induced inducible NO synthase (iNOS), and cyclooxygenase‐2 (COX‐2) expressions at messenger RNA and protein levels, and reversed the promoter activity of iNOS in RAW264.7 cells. Mechanistically, GEN‐27 abated LPS‐induced reactive oxygen species production, as well as mitigated LPS‐induced increase of caspase 1 activity and the protein levels of NOD‐like receptor 3 (NLRP3), anti‐apoptosis‐associated speck‐like protein‐containing a CRAD (ASC), and caspase 1 in RAW264.7 cells in a dose‐dependent manner. Similarly, GEN‐27 dose‐dependently weakened adenosine triphosphate‐induced NLRP3 and IL‐1β in RAW264.7 cells. In addition, GEN‐27 treatment significantly suppressed LPS‐induced phosphorylation of nuclear factor‐κB (NF‐κB) p65 and alleviated LPS‐induced increase of transcriptional activity of NF‐κB in RAW264.7 cells. In summary, these results revealed that GEN‐27 exhibited anti‐inflammatory effects by suppressing the activation of NLRP3 inflammasome and NF‐κB pathway, suggesting that GEN‐27 may be served as a promising therapeutic agent for the prevention and therapy of inflammatory‐associated diseases.