Chemical information of fucoxanthin-rich fraction (FxRF) from Sargassum fusiformis. (A) Extraction and isolation scheme for FxRF. (B) The HPLC chromatographs of Fx standard and FxRF. The mobile phases used in the isocratic elution consisted of eluent: 0.1% formic acid water and 95% methanol. The flow rate was 0.3 mL/min and UV detection was observed at 445 nm. (C) The mass spectrum of fucoxanthin from FxRF.

Efficacy of FxRF against inflammation induced by PM in RAW 264.7 macrophages (A), and analyses of RAW cell viability and intracellular ROS levels; (B) Western blot analyses of iNOS and COX-2 expressions; and (C) ELISA of PGE₂ and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α). Pre-seeded cells (1 × 10⁵ cells/mL) were treated with different FxRF concentrations after 24 h and stimulated with PM after 30 min. Cells were harvested after 24 h to measure inflammatory mediators (COX-2 and PGE₂) and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α). Graphical representations are means ± SE based on three replications. *p < 0.05 and **p < 0.01 indicate that the values were significantly different from those for the PM-treated group.

Efficacy of FxRF against inflammation induced by PM in HaCaT keratinocytes (A), and analyses of HaCaT cell viability and intracellular ROS levels; (B) Western blot analyses of COX-2 expressions; (C) levels of key molecular mediators in the MAPK pathways; and (D) ELISA of PGE2 and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α). Pre-seeded cells (1 × 105 cells/mL) were treated with different FxRF concentrations after 24 h and stimulated with PM after 30 min. Cells were harvested after 24 h to measure inflammatory mediators (COX-2 and PGE2) and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α). Apoptotic body formation was observed under a fluorescence microscope after (E) DCFH-DA treatment and (F) Hoechst 33,342 and PI staining. Graphical representations are means ± standard error (SE) based on three replications. *p < 0.05 and **p < 0.01 indicate that the values were significantly different from those for the PM-treated group..

Efficacy of FxRF against inflammation induced by PM in RAW 264.7 macrophages (A), and analyses of RAW cell viability and intracellular ROS levels; (B) Western blot analyses of iNOS and COX-2 expressions; and (C) ELISA of PGE₂ and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α). Pre-seeded cells (1 × 10⁵ cells/mL) were treated with different FxRF concentrations after 24 h and stimulated with PM after 30 min. Cells were harvested after 24 h to measure inflammatory mediators (COX-2 and PGE₂) and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α). Graphical representations are means ± SE based on three replications. *p < 0.05 and **p < 0.01 indicate that the values were significantly different from those for the PM-treated group.

Inflammatory stimulation of the RAW 264.7 macrophages by the culture medium of PM-induced HaCaT cells and the anti-inflammatory eff ;ects of FxRF: NO production and cytotoxicity (A), and analysis of iNOS and COX-2 levels (B) and inflammatory mediators (C), including tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6, and prostaglandin E2 (PGE₂). The HaCaT cells were pre-seeded in culture plates (1 × 10⁵ cells/mL), incubated for 24 h, and treated with diff ;erent concentrations of FxRF. After 1 h, the cells were treated with PM (125 μg/mL) and 24 h later, the culture media were treated to each pre-seeded RAW 264.7 macrophages culture well plates in real time. The evaluations were made after a 24 h. Experiments were carried out in triplicate, and the results are represented as means ± SE. Values are significantly diff ;erent from the positive control (PM treated group) at *p < 0.05 and **p < 0.001. H-PM: The cultured medium of PM-stimulated in keratinocytes.

Inflammatory stimulation of zebrafish larvae by PM and anti-inflammatory eff ;ects of FxRF: anti-inflammatory properties were evaluated by measuring NO and ROS production, and cell death in the zebrafish embryo model. Experiments were carried out in triplicate, and the results are represented as means ± SE. Values are significantly diff ;erent from the positive control (PM treated group) at *p < 0.05 and **p < 0.001.