Identification of Chemical Constituents and Inhibitory Effect of Ficus deltoidea Fraction against Lipopolysaccharide‑Induced Nuclear Factor‑Kappa B Inflammatory Pathway in Murine Macrophage 264.7 Cells

Objectives: The aim is to determine the anti‑inflammatory mechanism of various solvent fractions of F. deltoidea methanol extract against Lipopolysaccharide (LPS)‑induced nuclear factor‑kappa B (NF‑κB) inflammatory pathway in murine macrophage 264.7 cells and to identify the chemical constituents present in the active fraction. Materials and Methods: The effect of crude methanolic extract and its fractions (hexane, chloroform, ethyl acetate, and butanol) on murine macrophages against LPS‑induced pro‑inflammatory cytokines (interleukin [IL]‑Iβ, tumor necrosis factor alpha [TNF‑α], and IL‑6) and biomarkers were tested using enzyme‑linked immunosorbent assay and immunoblot analysis. The chemical constituents present in the active fraction were identified using liquid chromatography mass spectrometry and liquid chromatography tandem mass spectrometry analysis. Results: The findings indicated that among all the fractions, the ethyl acetate fraction of F. deltoidea substantially inhibits the LPS‑induced inflammatory mediators such as nitric oxide (NO) and pro‑inflammatory cytokine production including TNF‑α, IL‑6, and IL‑1 β in a dose‑dependent manner. The expression of inducible NO synthase, NO synthase, and cyclooxygenase‑2 were also effectively downregulated by the treatment of ethyl acetate fraction. Moreover, it also suppressed the expression of LPS‑induced NF‑κB translocation correlated with the inhibition of NF‑κB (inhibitor of kappa B alpha) degradation. The presence of bioactive phenolics, especially flavonoids such as catechin, vitexin, dodecadienyl coumaric acid, (epi)‑afzelechin‑(epi)‑catechin, genistein, and apigenin derivatives were identified in the ethyl acetate fraction of F. deltoidea. Conclusion: Overall, it has been recommended that the ethyl acetate fraction of F. deltoidea could be utilized as a potential natural anti‑inflammatory agent.