title: Anti-inflammatory activity of a sulfated polysaccharide isolated from an enzymatic digest of brown seaweedSargassum horneriin RAW 264.7 cells.
abstract: OBJECTIVE
  Sargassum horneriis an edible brown alga that grows in the subtidal zone as an annual species along the coasts of South Korea, China, and Japan. Recently, an extreme amount ofS. hornerimoved into the coasts of Jeju Island from the east coast of China, which made huge economic and environmental loss to the Jeju Island. Thus, utilization of this biomass becomes a big issue with the local authorities. Therefore, the present study was performed to evaluate the anti-inflammatory potential of crude polysaccharides (CPs) extracted fromS. horneriChina strain in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells.
  
  
  METHODS
  CPs were precipitated fromS. horneridigests prepared by enzyme assistant extraction using four food-grade enzymes (AMG, Celluclast, Viscozyme, and Alcalase). The production levels of nitric oxide (NO) and pro-inflammatory cytokines, including tumor necrosis factor (TNF)-α and interleukin (IL)-1β were measured by Griess assay and enzyme-linked immunosorbent assay, respectively. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), nuclear factor (NF)-κB, and mitogen-activated protein kinases (MAPKs) were measured by using western blot. The IR spectrums of the CPs were recorded using a fourier transform infrared spectroscopy (FT-IR) spectrometer.
  
  
  RESULTS
  The polysaccharides from the Celluclast enzyme digest (CCP) showed the highest inhibition of NO production in LPS-stimulated RAW 264.7 cells (IC50value: 95.7 µg/mL). Also, CCP dose-dependently down-regulated the protein expression levels of iNOS and COX-2 as well as the production of inflammatory cytokines, including TNF-α and IL-1β, compared to the only LPS-treated cells. In addition, CCP inhibited the activation of NF-κB p50 and p65 and the phosphorylation of MAPKs, including p38 and extracellular signal-regulated kinase, in LPS-stimulated RAW 264.7 cells. Furthermore, FT-IR analysis showed that the FT-IR spectrum of CCP is similar to that of commercial fucoidan.
  
  
  CONCLUSIONS
  Our results suggest that CCP has anti-inflammatory activities and is a potential candidate for the formulation of a functional food ingredient or/and drug to treat inflammatory diseases.
authors.affiliation: Laboratory of Marine Bioresource Technology, Department of Marine Life Science, School of Marine Biomedical Sciences, Jeju National University, 102 Jejudaehak-ro, Jeju 63243, Korea.
authors.forename: Kalu Kapuge Asanka
authors.initials: KK
authors.lastname: Sanjeewa
authors.affiliation: Laboratory of Marine Bioresource Technology, Department of Marine Life Science, School of Marine Biomedical Sciences, Jeju National University, 102 Jejudaehak-ro, Jeju 63243, Korea.
authors.forename: Ilekkuttige Priyan Shanura
authors.initials: IP
authors.lastname: Fernando
authors.affiliation: Laboratory of Marine Bioresource Technology, Department of Marine Life Science, School of Marine Biomedical Sciences, Jeju National University, 102 Jejudaehak-ro, Jeju 63243, Korea.
authors.forename: Eun-A
authors.initials: EA
authors.lastname: Kim
authors.affiliation: Department of Marine Bio Food Science, Chonnam National University, Yeosu 59626, Korea.
authors.forename: Ginnae
authors.initials: G
authors.lastname: Ahn
authors.affiliation: Department of Veterinary Medicine and Veterinary Medical Research Institute, Jeju National University, Jeju 63243, Korea.
authors.forename: Youngheun
authors.initials: Y
authors.lastname: Jee
authors.affiliation: Laboratory of Marine Bioresource Technology, Department of Marine Life Science, School of Marine Biomedical Sciences, Jeju National University, 102 Jejudaehak-ro, Jeju 63243, Korea.
authors.forename: You-Jin
authors.initials: YJ
authors.lastname: Jeon
keywords: Macrophage; Sargassum horneri; inflammation; sulfated polysaccharide