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Anti-inflammatory Effects of Ethanolic Extracts from Codium fragile on LPS-Stimulated RAW 264.7 Macrophages via Nuclear Factor kappaB Inactivation

  • Yoon, Ho-Dong (Food and Safety Research Division, National Fisheries Research and Development Institute) ;
  • Jeong, Eun-Ji (Department of Food Science and Nutrition, Pukyong National University) ;
  • Choi, Ji-Woong (Department of Food Science and Nutrition, Pukyong National University) ;
  • Lee, Min-Sup (Department of Food Science and Nutrition, Pukyong National University) ;
  • Park, Myoung-Ae (Pathology Division, National Fisheries Research and Development Institute) ;
  • Yoon, Na-Young (Food and Safety Research Division, National Fisheries Research and Development Institute) ;
  • Kim, Yeon-Kye (Food and Safety Research Division, National Fisheries Research and Development Institute) ;
  • Cho, Deuk-Moon (Department of Food and Nutrition, Dong-Pusan College) ;
  • Kim, Jae-Il (Department of Food Science and Nutrition, Pukyong National University) ;
  • Kim, Hyeung-Rak (Department of Food Science and Nutrition, Pukyong National University)
  • Received : 2011.09.15
  • Accepted : 2011.11.10
  • Published : 2011.12.31

Abstract

Bacterial lipopolysaccharide (LPS) induces expression of pro-inflammatory cytokines and enzymes producing nitric oxide (NO) and prostaglandins (PGs) in immune cells. This process is mediated by the activation of nuclear factor kappaB (NF-${\kappa}B$). In this study, we investigated the anti-inflammatory characteristics of Codium fragile ethanolic extract (CFE) mediated by the regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) using LPS-stimulated murine macrophage RAW 264.7 cells. CFE significantly inhibited LPS-induced NO and $PGE_2$ production in a dose-dependent manner and suppressed the expression of iNOS and COX-2 proteins in LPS-stimulated RAW 264.7 cells with no cytotoxicity. Pro-inflammatory cytokines, such as interleukin (IL)-$1{\beta}$, IL-6, and tumor necrosis factor-${\alpha}$, were significantly reduced by treatment of CFE in LPS-stimulated RAW 264.7 cells. CFE inhibited the promoter activity of (NF)-${\kappa}B$ in LPS-stimulated macrophages. Treatment with CFE suppressed translocation of the NF-${\kappa}B$ p65 subunit by preventing proteolytic degradation of inhibitor of ${\kappa}B-{\alpha}$. These results indicate that the CFE-mediated inhibition of NO and $PGE_2$ production in LPS-stimulated RAW 264.7 cells is mediated through the NF-${\kappa}B$-dependent transcriptional downregulation of iNOS and COX-2, suggesting the potential of CFE as a nutraceutical with anti-inflammatory activity.

Keywords

References

  1. Abad MJ, Bedoya LM and Bermejo P. 2008. Natural marine anti-inflammatory products. Mini Rev Med Chem 8, 740-754. https://doi.org/10.2174/138955708784912148
  2. Beutler B and Cerami A. 1989. The biology of cachectin/TNF--a primary mediator of the host response. Annu Rev Immunol 7, 625-655. https://doi.org/10.1146/annurev.iy.07.040189.003205
  3. Blunt JW, Copp BR, Munro MH, Northcote PT and Prinsep MR. 2010. Marine natural products. Nat Prod Rep 27, 165-237. https://doi.org/10.1039/b906091j
  4. Chen Z, Hagler J, Palombella VJ, Melandri F, Scherer D, Ballard D and Maniatis T. 1995. Signal-induced site-specific phosphorylation targets $I{\kappa}B{\alpha}$ to the ubiquitin-proteasome pathway. Genes Dev 9, 1586-1597. https://doi.org/10.1101/gad.9.13.1586
  5. Chung EY, Kim BH, Hong JT, Lee CK, Ahn B, Nam SY, Han SB and Kim Y. 2011. Resveratrol down-regulates interferon-$\gamma$-inducible inflammatory genes in macrophages: molecular mechanism via decreased STAT-1 activation. J Nutr Biochem 22, 902-909. https://doi.org/10.1016/j.jnutbio.2010.07.012
  6. D'Acquisto F, Iuvone T, Rombola L, Sautebin L, Di Rosa M and Carnuccio R. 1997. Involvement of $NF-{\kappa}B$ in the regulation of cyclooxygenase-2 protein expression in LPS-stimulated J774 macrophages. FEBS Lett 418, 175-178. https://doi.org/10.1016/S0014-5793(97)01377-X
  7. Dinarello CA. 1999. Cytokines as endogenous pyrogens. J Infect Dis 179(Suppl 2), S294-S304. https://doi.org/10.1086/513856
  8. Ganesan P, Matsubara K, Ohkubo T, Tanaka Y, Noda K, Sugawara T and Hirata T. 2010. Anti-angiogenic effect of siphonaxanthin from green alga, Codium fragile. Phytomedicine 17, 1140-1144. https://doi.org/10.1016/j.phymed.2010.05.005
  9. Guha M and Mackman N. 2001. LPS induction of gene expression in human monocytes. Cell Signal 13, 85-94. https://doi.org/10.1016/S0898-6568(00)00149-2
  10. Heo SJ, Yoon WJ, Kim KN, Ahn GN, Kang SM, Kang DH, Affan A, Oh C, Jung WK and Jeon YJ. 2010. Evaluation of anti-inflammatory effect of fucoxanthin isolated from brown algae in lipopolysaccharide-stimulated RAW 264.7 macrophages. Food Chem Toxicol 48, 2045-2051. https://doi.org/10.1016/j.fct.2010.05.003
  11. Jin M, Suh SJ, Yang JH, Lu Y, Kim SJ, Kwon S, Jo TH, Kim JW, Park YI, Ahn GW, Lee CK, Kim CH, Son JK, Son KH and Chang HW. 2010. Anti-inflammatory activity of bark of Dioscorea batatas DECNE through the inhibition of iNOS and COX-2 expressions in RAW264.7 cells via $NF-{\kappa}B$and ERK1/2 inactivation. Food Chem Toxicol 48, 3073-3079. https://doi.org/10.1016/j.fct.2010.07.048
  12. Jung WK, Ahn YW, Lee SH, Choi YH, Kim SK, Yea SS, Choi I, Park SG, Seo SK, Lee SW and Choi IW. 2009. Ecklonia cava ethanolic extracts inhibit lipopolysaccharide-induced cyclooxygenase-2 and inducible nitric oxide synthase expression in BV2 microglia via the MAP kinase and $NF-{\kappa}B$ pathways. Food Chem Toxicol 47:410-417. https://doi.org/10.1016/j.fct.2008.11.041
  13. Kim AR, Shin TS, Lee MS, Park JY, Park KE, Yoon NY, Kim JS, Choi JS, Jang BC, Byun DS, Park NK and Kim HR. 2009. Isolation and identification of phlorotannins from Ecklonia stolonifera with antioxidant and anti-inflammatory properties. J Agric Food Chem 57, 3483-3489. https://doi.org/10.1021/jf900820x
  14. Kim EY and Moudgil KD. 2008. Regulation of autoimmune inflammation by pro-inflammatory cytokines. Immunol Lett 120, 1-5. https://doi.org/10.1016/j.imlet.2008.07.008
  15. Kim MM and Kim SK. 2010. Effect of phloroglucinol on oxidative stress and inflammation. Food Chem Toxicol 48, 2925-2933. https://doi.org/10.1016/j.fct.2010.07.029
  16. Kim YC, An RB, Yoon NY, Nam TJ and Choi JS. 2005. Hepatoprotective constituents of the edible brown alga Ecklonia stolonifera on tacrine-induced cytotoxicity in Hep G2 cells. Arch Pharm Res 28, 1376-1380. https://doi.org/10.1007/BF02977904
  17. Lebovic DI, Bentzien F, Chao VA, Garrett EN, Meng YG and Taylor RN. 2000. Induction of an angiogenic phenotype in endometriotic stromal cell cultures by interleukin-1$\beta$. Mol Hum Reprod 6, 269-275. https://doi.org/10.1093/molehr/6.3.269
  18. Lee JB, Ohta Y, Hayashi K and Hayashi T. 2010. Immunostimulating effects of a sulfated galactan from Codium fragile. Carbohydr Res 345, 1452-1454. https://doi.org/10.1016/j.carres.2010.02.026
  19. Li Q and Verma IM. 2002. $NF-{\kappa}B$ regulation in the immune system. Nat Rev Immunol 2, 725-734. https://doi.org/10.1038/nri910
  20. Libby P. 2006. Inflammation and cardiovascular disease mechanisms. Am J Clin Nutr 83, 456S-460S.
  21. Makarov SS. 2001. $NF-{\kappa}B$ in rheumatoid arthritis: a pivotal regulator of inflammation, hyperplasia, and tissue destruction. Arthritis Res 3, 200-206. https://doi.org/10.1186/ar300
  22. Marks-Konczalik J, Chu SC and Moss J. 1998. Cytokine-mediated transcriptional induction of the human inducible nitric oxide synthase gene requires both activator protein 1 and nuclear factor ${\kappa}B$-binding sites. J Biol Chem 273, 22201-22208. https://doi.org/10.1074/jbc.273.35.22201
  23. Nathan C. 1992. Nitric oxide as a secretory product of mammalian cells. FASEB J 6, 3051-3064.
  24. Ohta Y, Lee JB, Hayashi K and Hayashi T. 2009. Isolation of sulfated galactan from Codium fragile and its antiviral effect. Biol Pharm Bull 32, 892-898. https://doi.org/10.1248/bpb.32.892
  25. Packard RR and Libby P. 2008. Inflammation in atherosclerosis: from vascular biology to biomarker discovery and risk prediction. Clin Chem 54, 24-38.
  26. Pan MH, Hong HM, Lin CL, Jhang AZ, Tsai JH, Badmaev V, Nagabhushanam K, Ho CT and Chen WJ. 2011. Se-methylselenocysteine inhibits lipopolysaccharide-induced $NF-{\kappa}B$ activation and iNOS induction in RAW 264.7 murine macrophages. Mol Nutr Food Res 55, 723-732. https://doi.org/10.1002/mnfr.201000481
  27. Shin JS, Park YM, Choi JH, Park HJ, Shin MC, Lee YS and Lee KT. 2010. Sulfuretin isolated from heartwood of Rhus verniciflua inhibits LPS-induced inducible nitric oxide synthase, cyclooxygenase-2, and pro-inflammatory cytokines expression via the downregulation of $NF-{\kappa}B$ in RAW 264.7 murine macrophage cells. Int Immunopharmacol 10, 943-950. https://doi.org/10.1016/j.intimp.2010.05.007
  28. Solinas G, Marchesi F, Garlanda C, Mantovani A and Allavena P. 2010. Inflammation-mediated promotion of invasion and metastasis. Cancer Metastasis Rev 29, 243-248. https://doi.org/10.1007/s10555-010-9227-2
  29. Tseng CK and Chang CF. 1984. Chienese seaweeds in herbal medicine. Hydrobiologia 116/117, 152-154. https://doi.org/10.1007/BF00027655
  30. Vane JR, Mitchell JA, Appleton I, Tomlinson A, Bishop-Bailey D, Croxtall J and Willoughby DA. 1994. Inducible isoforms of cyclooxygenase and nitric-oxide synthase in inflammation. Proc Natl Acad Sci U S A 91, 2046-2050. https://doi.org/10.1073/pnas.91.6.2046
  31. Woo MN, Jeon SM, Shin YC, Lee MK, Kang MA and Choi MS. 2009. Anti-obese property of fucoxanthin is partly mediated by altering lipid-regulating enzymes and uncoupling proteins of visceral adipose tissue in mice. Mol Nutr Food Res 53, 1603-1611. https://doi.org/10.1002/mnfr.200900079
  32. Xie QW, Whisnant R and Nathan C. 1993. Promoter of the mouse gene encoding calcium-independent nitric oxide synthase confers inducibility by interferon $\gamma$ and bacterial lipopolysaccharide. J Exp Med 177, 1779-1784. https://doi.org/10.1084/jem.177.6.1779
  33. Yoshimura A. 2006. Signal transduction of inflammatory cytokines and tumor development. Cancer Sci 97, 439-447. https://doi.org/10.1111/j.1349-7006.2006.00197.x
  34. Zhang G and Ghosh S. 2000. Molecular mechanisms of $NF-{\kappa}B$ activation induced by bacterial lipopolysaccharide through Toll-like receptors. J Endotoxin Res 6, 453-457.

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