Anti‑inflammatory Effect of Procumbenoside B from Justicia spicigera on Lipopolysaccharide‑Stimulated RAW 264.7 Macrophages and Zebrafish Model

Background: Justicia spicigera is widely used in the Mexican traditional medicine for the treatment of inflammation. Objectives: The purpose of this study was to investigate the anti‑inflammatory effect of procumbenoside B (PB) from J. spicigera on pro‑inflammatory mediators. Materials and Methods: Lipopolysaccharide (LPS)‑stimulated RAW264.7 macrophages and Zebrafish model were used to assess the potential anti‑inflammatory effects of PB; its structure was elucidated on the basis of spectroscopic data. The production of inflammatory mediators such as interferon‑β, prostaglandin E2, inducible nitric oxide synthase (iNOS), nuclear factor‑kappa B (NF‑κB) p65, interleukin‑6 (IL‑6), IL‑1β, IL‑12, cyclooxygenase (COX‑2), tumor necrosis factor‑α, and anti‑inflammatory IL‑10 was measured using enzyme‑linked immunosorbent assay. NO production was measured using Griess reagent. Results: In LPS‑induced‑inflammatory response in RAW264.7 macrophage cells, PB strongly inhibited secretion of all pro‑inflammatory mediators test and increased the production of IL‑10 and blockade of NF‑κB. In addition, PB suppressed LPS‑stimulated nitric oxide and reactive oxygen species generation in a zebrafish model. Conclusion: These results indicated that the anti‑inflammatory effects of PB may be attributed to the downregulation of iNOS and COX‑2 through the suppression of NF‑κB signaling pathway in RAW264.7 macrophages.