Tuberatolide B Protects LPS-stimulated Inflammatory Response in RAW264.7 cells and Zebrafish Model through Inhibiting MAPKs and NF-kB Signalings

Abstract

Background : Brown algae are known to contain rich anti-inflammatory compounds. The objective of this study was to identify anti-inflammatory compound in the Sargassum macrocarpum and anti-inflammatory effect of tuberatolide B (TTB) was investigated. Methods: RAW 264.7 cells were seeded on the well plates. After 16 h, cells were treated with TTB incubated and subsequently treated with LPS. After 24 h, we measured cell viability and NO production using MTT or griess regents, and inflammatory cytokines using ELISA kit as well as signaling pathways by western blotting in in vitro cell experiment. Zebrafish embryos were transferred to the well plate. At 7-9 hpf, TTB and LPS were added to the well. We are confirmedNO production and cell death using DAF-FM-DA or acridine orange solution in in vivo zebrafish experiment. Results : TTB significantly suppressed the production of NO and pro-inflammatory cytokines such as interleukin (IL)-6 and IL-1β. In addition, the mechanism involved in the inhibition of the inflammatory effect of LPS by TTB was investigated. The results showed that TTB aforementioned this effect through NF-κB and MAPKs phosphorylation pathways. We also assessed the anti-inflammatory activity of TTB using in vivo zebrafish model. In LPS-stimulated zebrafish, TTB enhanced survival and significantly inhibited the NO production and mRNA expression of inducible NO synthase.