Journal of Veterinary Clinics (한국임상수의학회지)

The Korean Society of Veterinary Clinics (한국임상수의학회)
권호 표지
DOI QR코드

DOI QR Code

Effects of Fucoidan on Nitric Oxide Production and Activator Protein-1 Activation in Lipopolysaccharide-Stimulated Porcine Peripheral Blood Mononuclear Cells

LPS로 자극한 돼지 말초혈액 단핵구세포의 Nitric Oxide (NO) 생산 및 Activator Protein-1 (AP-1) 활성화에 있어 Fucoidan의 효과

  • Park, Jongchan (Department of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University) ;
  • Ahn, Changhwan (Department of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University) ;
  • Kang, Byeong-Teck (Department of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University) ;
  • Kang, Ji-Houn (Department of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University) ;
  • Jeung, Eui-Bae (Department of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University) ;
  • Yang, Mhan-Pyo (Department of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University)
  • Accepted : 2015.08.05
  • Published : 2015.08.31
Download PDF
⟨ Previous Next ⟩

Abstract

Fucoidan which is sulfated polysaccharide extracted from brown seaweed has a wide variety of internal biological activities. The objectives of this study were to examine the effect of fucoidan on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated porcine peripheral blood mononuclear cells (PBMCs) and to investigate whether this effect is involved in the expression of inducible nitric oxide synthase (iNOS) and the activation of activator portein-1 (AP-1). The levels of NO production and AP-1 activity in the culture supernatants from porcine PBMCs were measured by the enzyme-linked immunosorbent assay and the levels of iNOS and AP-1 mRNA were determined by real time polymerase chain reaction. Fucoidan in LPS-naïve PBMCs has no effects on the production of NO and activity of AP-1. Expressions of iNOS and AP-1 mRNA in LPS-naïve PBMCs were also not affected by treatment of fucoidan. However, NO production, AP-1 activity and expressions of iNOS and AP-1 mRNA were dramatically increased in PBMCs stimulated with LPS. Enhancing effects of NO production and AP-1 activity in PBMCs induced by LPS were reduced by addition of fucoidan. Fucoidan also inhibited an increase in expressions of iNOS and AP-1 mRNA in LPS-stimulated PBMCs. These results suggested that fucoidan exerts anti-inflammatory effect by down-regulating production of NO via suppressing expression of iNOS and activity of AP-1 in LPS-stimulated porcine PBMCs.

Fucoidan은 갈조류로부터 추출되는 황산다당류로 다양한 생리학적 활성을 갖고 있다. 본 연구의 목적은 LPS로 자극한 돼지 말초혈액 단핵구세포(PBMCs)의 NO 생산에 있어 fucoidan의 효과를 검토하고, 이러한 효과가 iNOS의 발현과 AP-1의 활성화와 관련이 있는지를 조사하는데 있다. LPS 무처치 돼지 PBMCs에서 fucoidand의 처리는 NO 생산과 AP-1활성에 대해 효과를 보이지 않았다. 또한 iNOS와 AP-1의 mRNA 발현도 fucoidan 처치에 의해 영향을 받지 않았다. 그러나 LPS로 자극한 PBMCs에서는 NO 생산과 AP-1의 활성 그리고 iNOS와 AP-1의 mRNA 발현이 현저하게 증가하였다. 이와 같은 LPS에 의한 돼지 PBMCs의 NO 생산과 AP-1 활성증가는 fucoidan 첨가에 의해 감소되었다. 또한 fucoidan은 LPS에 의한 iNOS와 AP-1의 mRNA 발현증가도 억제시켰다. 이상의 결과는 fucoidan이 LPS 자극 돼지 PBMCs에서 iNOS 발현과 AP-1 활성의 억제와 함께 NO 생산을 하향 조절함으로써 항염증효과를 나타내는 것으로 사료되었다.

Keywords

References

  1. Baba M, Snoeck R, Pauwels R, De Clercq E. Sulfated polysaccharides are potent and selective inhibitors of various enveloped viruses, including herpes simplex virus, cytomegalovirus, vesicular stomatitis virus, and human immunodeficiency virus. Antimicrob Agents Chemother 1988; 32: 1742-1745. https://doi.org/10.1128/AAC.32.11.1742
  2. Backstrom L, Hoefling DC, Morkoc AC, Cowart RP. Effect of atrophic rhinitis on growth rate in Illinois swine herds. J AVMA 1985; 187: 712-715.
  3. Berteau O, Mulloy B. Sulfated fucans, fresh perspectives: structures, functions, and biological properties of sulfated fucans and an overview of enzymes active toward this class of polysaccharide. Glycobiology 2003; 13: 29R-40R. https://doi.org/10.1093/glycob/cwg058
  4. Bilan MI, Grachev AA, Ustuzhanina NE, Shashkov AS, Nifantiev NE, Usov AI. Structure of a fucoidan from the brown seaweed Fucus evanescens C.Ag. Carbohydr Res 2002; 337: 719-730. https://doi.org/10.1016/S0008-6215(02)00053-8
  5. Block ML, Zecca L, Hong JS. Microglia-mediated neurotoxicity: uncovering the molecular mechanisms. Nature 2007; 8: 57-69.
  6. Chen YC, Shen SC, Lee WR, Hou WC, Yang LL, Lee TJ. Inhibition of nitric oxide synthase inhibitors and lipopolysaccharide induced inducible NOS and cyclooxygenase-2 gene expressions by rutin, quercetin, and quercetin pentaacetate in RAW 264.7 macrophages. J Cell Biochem 2001; 82: 537-548. https://doi.org/10.1002/jcb.1184
  7. Choi RJ, Chun J, Khan S, Kim YS. Desoxyrhapontigenin, a potent anti-inflammatory phytochemical, inhibits LPSinduced inflammatory responses via suppressing NF-${\kappa}B$ and MAPK pathways in RAW 264.7 cells. Int J Immunopharmacol 2014; 18: 182-190. https://doi.org/10.1016/j.intimp.2013.11.022
  8. Chun J, Choi RJ, Khan S, Lee DS, Kim YC, Nam YJ, Kim YS. Alantolactone suppresses inducible nitric oxide synthase and cyclooxygenase-2 expression by down-regulating NF-${\kappa}B$, MAPK and AP-1 via the MyD88 signaling pathway in LPS-activated RAW 264.7 cells. Int J Immunopharmacol 2012; 14: 375-383. https://doi.org/10.1016/j.intimp.2012.08.011
  9. Cui YQ, Zhang LJ, Zhang T, Luo DZ, Jia YJ, Guo ZX, Wang XM. Inhibitory effect of fucoidan on nitric oxide production in lipopolysaccharide-activated primary microglia. Clin Exp Pharmacol Physiol 2010; 37: 422-428. https://doi.org/10.1111/j.1440-1681.2009.05314.x
  10. Cumashi A, Ushakova NA, Preobrazhenskaya ME, D'Incecco A, Piccoli A, Totani L, Nifantiev NE. A comparative study of the anti-inflammatory, anticoagulant, antiangiogenic, and antiadhesive activities of nine different fucoidans from brown seaweeds. Glycobiology 2007; 17: 541-552. https://doi.org/10.1093/glycob/cwm014
  11. Dauphinee SM, Karsan A. Lipopolysaccharide signaling in endothelial cells. Lab Invest 2005; 86: 9-22.
  12. DeGroote MA, Fang FC. Antimicrobial properties of nitric oxide. In: Nitric oxide and infection, New York: Kluwer Academic Publishers, 2002: 231-261.
  13. Feng GJ, Goodridge HS, Harnett MM, Wei XQ, Nikolaev AV, Higson AP, Liew FY. Extracellular signal-related kinase (ERK) and p38 mitogen-activated protein (MAP) kinases differentially regulate the lipopolysaccharide-mediated induction of inducible nitric oxide synthase and IL-12 in macrophages: Leishmania phosphoglycans subvert macrophage IL-12 production by targeting ERK MAP kinase. J Immunol 1999; 163: 6403-6412.
  14. Irhimeh MR, Fitton JH, Lowenthal RM. Pilot clinical study to evaluate the anticoagulant activity of fucoidan. Blood Coagul Fibrin 2009; 20: 607-610. https://doi.org/10.1097/MBC.0b013e32833135fe
  15. Iwakiri Y, Sampson DA, Allen KGD. Suppression of cyclooxygenase-2 and inducible nitric oxide synthase expression by conjugated linoleic acid in murine macrophages. Prostag Leukotr Ess 2002; 67: 435-443. https://doi.org/10.1054/plef.2002.0454
  16. Kikuchi T, Hagiwara K, Honda Y, Gomi K, Kobayashi T, Takahashi H, Nukiwa T. Clarithromycin suppresses lipopolysaccharide-induced interleukin-8 production by human monocytes through AP-1 and NF-${\kappa}B$ transcription factors. J Antimicrob Chemother 2002; 49: 745-755. https://doi.org/10.1093/jac/dkf008
  17. Kim DI, Kim KH, Kang JH, Jung EM, Kim SS, Jeung EB, Yang MP. Trans-10, cis-12-conjugated linoleic acid modulates NF-${\kappa}B$ activation and TNF-$\alpha$ production in porcine peripheral blood mononuclear cells via a $PPAR\gamma$-dependent pathway. Brit J Nutr 2011; 105: 1329-1336. https://doi.org/10.1017/S000711451000499X
  18. Koyanagi S, Tanigawa N, Nakagawa H, Soeda S, Shimeno H. Oversulfation of fucoidan enhances its anti-angiogenic and antitumor activities. Biochem Pharmacol 2003; 65: 173-179. https://doi.org/10.1016/S0006-2952(02)01478-8
  19. Kroncke KD, Fehsel K, Kolb-Bachofen V. Inducible nitric oxide synthase and its product nitric oxide, a small molecule with complex biological activities. Biol Chem Hoppe Seyler 1995; 376: 327-343.
  20. Kwon DJ, Ju SM, Youn GS, Choi SY, Park J. Suppression of iNOS and COX-2 expression by flavokawain A via blockade of NF-${\kappa}B$ and AP-1 activation in RAW 264.7 macrophages. Food Chem Toxicol 2013; 58: 479-486 https://doi.org/10.1016/j.fct.2013.05.031
  21. Li B, Lu F, Wei X, Zhao R. Fucoidan: structure and bioactivity. Molecules 2008; 13: 1671-1695. https://doi.org/10.3390/molecules13081671
  22. Mankan AK, Lawless MW, Gray SG, Kelleher D, McManus R. NF-${\kappa}B$ regulation: the nuclear response. J Cell Mol Med 2009; 13: 631-643. https://doi.org/10.1111/j.1582-4934.2009.00632.x
  23. Murphy S. Production of nitric oxide by glial cells: regulation and potential roles in the CNS. Glia 2000; 29: 1-13. https://doi.org/10.1002/(SICI)1098-1136(20000101)29:1<1::AID-GLIA1>3.0.CO;2-N
  24. Nakamura T, Suzuki H, Wada Y, Kodama T, Doi T. Fucoidan induces nitric oxide production via p38 mitogenactivated protein kinase and NF-${\kappa}B$-dependent signaling pathways through macrophage scavenger receptors. Biochem Biophys Res Commun 2006; 343: 286-294. https://doi.org/10.1016/j.bbrc.2006.02.146
  25. Nathan C, Shiloh MU. Reactive oxygen and nitrogen intermediates in the relationship between mammalian hosts and microbial pathogens. Proc Natl Acad Sci USA 2000; 97: 8841-8848. https://doi.org/10.1073/pnas.97.16.8841
  26. Noman AS, Koide N, Khuda II, Dagvadorj J, Tumurkhuu G, Naiki Y, Komatsu T, Yoshida T, Yokochi T. Thalidomide inhibits lipopolysaccharide-induced nitric oxide production and prevents lipopolysaccharide-mediated lethality in mice. FEMS Immunol Med Microbiol 2009; 56: 204-211. https://doi.org/10.1111/j.1574-695X.2009.00567.x
  27. Ohshima H, Bartsch H. Chronic infections and inflammatory processes as cancer risk factors: possible role of nitric oxide in carcinogenesis. Mutat Res 1994; 305: 253-264. https://doi.org/10.1016/0027-5107(94)90245-3
  28. Pampusch MS, Bennaars AM, Harsch S, Murtaugh MP. Inducible nitric oxide synthase expression in porcine immune cells. Vet Immunol Immunopathol 1998; 61: 279-289. https://doi.org/10.1016/S0165-2427(97)00139-6
  29. Park HY, Han MH, Park C, Jin CY, Kim GY, Choi IW, Choi YH. Anti-inflammatory effects of fucoidan through inhibition of NF-${\kappa}B$, MAPK and Akt activation in lipopolysaccharide-induced BV2 microglia cells. Food Chem Toxicol 2011; 49: 1745-1752. https://doi.org/10.1016/j.fct.2011.04.020
  30. Sarkar FH, Li Y, Wang Z, Kong D. NF-${\kappa}B$ signaling pathway and its therapeutic implications in human diseases. Int Rev Immunol 2008; 27: 293-319. https://doi.org/10.1080/08830180802276179
  31. Shan J, Fu J, Zhao Z, Kong X, Huang H, Luo L, Yin Z. Chlorogenic acid inhibits lipopolysaccharide-induced cyclooxygenase-2 expression in RAW264. 7 cells through suppressing NF-${\kappa}B$ and JNK/AP-1 activation. Int Immunopharmacol 2009; 9: 1042-1048. https://doi.org/10.1016/j.intimp.2009.04.011
  32. Shaulian E, Karin M. AP-1 in cell proliferation and survival. Oncogene 2001; 20: 2390-2400. https://doi.org/10.1038/sj.onc.1204383
  33. Sorensen NS, Skovgaard K, Heegaard PM. Porcine blood mononuclear cell cytokine responses to PAMP molecules: comparison of mRNA and protein production. Vet Immunol Immunopathol 2011; 139: 296-302. https://doi.org/10.1016/j.vetimm.2010.10.016
  34. Yang JW, Yoon SY, Oh SJ, Kim SK, Kang KW. Bifunctional effect of fucoidan on the expression of inducible nitric oxide synthase. Biochem Biophys Res Commun 2006; 346: 345-350. https://doi.org/10.1016/j.bbrc.2006.05.135
  35. Yokota T, Nagashima M, Ghazizadeh M, Kawanami O. Increased effect of fucoidan on lipoprotein lipase secretion in adipocytes. Life Sci 2009; 84: 523-529. https://doi.org/10.1016/j.lfs.2009.01.020

Cited by

  1. Effect of trans-10, cis-12 Conjugated Linoleic Acid on Production of Prostaglandin E2, Cyclooxygenase-2 and 5-lipoxygenase in Lipopolysaccharide-Stimulated Porcine Peripheral Blood Mononucl vol.33, pp.4, 2015, https://doi.org/10.17555/jvc.2016.08.33.4.194
  2. Fucoidan Suppresses Prostaglandin E2 Production and Akt Activation in Lipopolysaccharide-Stimulated Porcine Peripheral Blood Mononuclear Cells vol.34, pp.3, 2015, https://doi.org/10.17555/jvc.2017.06.34.3.172