Eupatorin interferes with NLRP3 inflammasome activation via modulation of NF-κB signaling pathway in LPS induced macrophages.

NOD-like cytosolic receptors (NLRs) are crucial in both nonspecific and specific immunity. Among them, the NLRP3 (NOD-like receptor family pyrin domain-containing 3) gains clinical attention, where its hyperactivation causes inflammatory, autoimmune, and metabolic illnesses. We investigated the effects of the flavone eupatorin (3', 5-Dihydroxy 4', 6, 7-trimethoxy flavone) on the NLRP3 inflammasome and associated proteins utilizing in vitro and in silico approaches. Eupatorin modulates the NLRP3 and associated molecules' expression in lipopolysaccharide (LPS)-primed, adenosine 5'-triphosphate (ATP)-stimulated THP-1 cells through downregulation of NF-ĸB phosphorylation, with promising effects as observed with standard tranilast. Additionally, the suppression of zymosan-induced intracellular oxidative stress and LPS-induced nitric oxide (NO) revealed anti-inflammatory effects. The strong binding affinity of -8.276 kcal/mol of the compound with NLRP3 was observed at the ATP binding pocket as shown by the standard tranilast, -7.834 kcal/mol, where interaction with NF-kB also provides comparable results with binding affinities of -3.93 and - 3.17 kcal/mol, respectively. The in silico results support the in vitro findings. Eupatorin appears to be a potential medication option for NLRP3-driven inflammatory and metabolic diseases and deserves further development as an inflammasome inhibitor.