Anti-inflammatory Effects of Enzymatic Extract from Ecklonia cava on TPA-induced Ear Skin Edema

  • Ahn, Ginnae (Department of Marine Life Science, Cheju National University) ;
  • Park, Eun-Jin (College of Veterinary Medicine and Institute of Medical Science, Cheju National University) ;
  • Kim, Dae-Seung (Department of Marine Life Science, Cheju National University) ;
  • Jeon, You-Jin (Faculty of Marine Medicine Science, Cheju National University) ;
  • Shin, Tae-Kyun (College of Veterinary Medicine and Institute of Medical Science, Cheju National University) ;
  • Park, Jae-Woo (Department of Nuclear and Energy Engineering, Cheju National University) ;
  • Woo, Ho-Chun (College of Veterinary Medicine and Institute of Medical Science, Cheju National University) ;
  • Lee, Ki-Wan (Department of Marine Life Science, Cheju National University) ;
  • Jee, Young-Heun (College of Veterinary Medicine and Institute of Medical Science, Cheju National University)
  • Published : 2008.08.31


Anti-inflammatory potential of the enzymatic extract prepared by Kojizyme (ECK), a component of brown seaweeds Ecklonia cava (Alariaceae, Phaeophyta) in vivo was investigated. For the application of mouse ear edema model, 12-O-tetradecanoylphorbol acetate (TPA) was used, a topical inducer of a long-lasting inflammatory response. Our results demonstrated that ECK inhibited ear edema when topically applied to mouse ear skin. In histological evaluation, the inhibition activity of ECK on TPA-induced inflammation is similar to that of dexamethasone, although less strong. In addition, the mRNA expression levels of IL-$1{\beta}$, IFN-$\gamma$, TNF-$\alpha$, and cyclooxygenase-2 (COX2) and the immunoreactivity to inducible nitric oxide synthase (iNOS) and COX2 expressed mainly in inflammatory cells were down-regulated by ECK. These results indicate that ECK has anti-inflammatory effects through the inhibition of Th1 cytokines and 2 inducers of inflammation in TPA-induced ear skin edema.



  1. Hashim MA, Chu KH. Biosorption of cadmium by brown, green, and red seaweeds. Chem. Eng. J. 97: 249-255 (2004)
  2. Siriwardhana N, Jeon YJ, Kim SH, Ha JH, Heo SJ, Lee KW. Enzymatic hydrolysis for effective extraction of antioxidative compounds from Hizikia fusiformis. Algae 19: 59-68 (2004)
  3. Ruperez P, Saura-Calixto F. Diatary fibers and physiochemical properties of edible seaweeds. Eur. Food Res. Technol. 212: 349-354 (2001)
  4. Heo SJ, Park EJ, Lee KW, Jeon YJ. Antioxidative effect of proteolytic hydrolysates from Ecklonia cava on radical scavenging using ESR and $H_2O_2$-induced DNA damage. Food Sci. Biotechnol. 14: 614-620 (2005)
  5. Kang KA, Lee KH, Chae SW, Koh YS, Yoo BS, Kim JH, Ham YM, Baik JS, Lee NH, Hyun JW. Triphlorethol-A from Ecklonia cava protects V79-4 lung fibroblasts against hydrogen peroxide induced cell damage. Free Radical Res. 39: 883-892 (2005)
  6. Kang KA, Lee KH, Chae SW, Zhang R, Jung MS, Lee YG, Kim SY, Kim HS. Eckol isolated from Ecklonia cava attenuates oxidative stress induced cell damage in lung fibroblast cells. FEBS Lett. 579: 6295-6304 (2005)
  7. Maruyama M, Tamauchi H, Hasimoto M, Nakano T. Antitumor activity and immune response of Mekabu fucoidan extracted from Sporophyll of Undaria pinnatifida. In Vivo 17: 245-249 (2003)
  8. Kim JA, Lee JM, Shin DB, Lee NH. The antioxidant activity and tyrosinase inhibitory activity of phloro-tannins in Ecklonia cava. Food Sci. Biotechnol. 13: 476-480 (2004)
  9. Fukuyama Y, Kodama M, Miura I, Kinzyo Z, Kido M, Nakayama Y, Takahashi H. Structure of an anti-plasmin inhibitor, eckol, isolated from the brown algae Ecklonia kurome Okamura and inhibitory activities of its derivatives on plasma plasmin inhibitors. Chem. Pharm. Bull. 37: 349-353 (1989)
  10. Nagayama K, Iwamura Y, Shibata T, Hirayama I, Nakamura T. Bactericidal activity of phlorotannins from the brown algae Ecklonia kurome. J. Antimicrob. Chemother. 50: 889-893 (2002)
  11. Lee JH, Kim ND, Choi JS, Kim YJ, Moon YH, Lim SY, Park KY. Inhibitory effects of the methanolic extract of an edible brown algae, Ecklonia stolonifera and its component, phlorglucinol on aflatoxin B1 mutagenicity in vitro (Ames test) and on benzo(a)pyrene or Nmethyl N-nitrosourea clastogenicity in vivo (mouse micronucleus test). Nat. Prod. Sci. 4: 105-114 (1998)
  12. Athukorala Y, Jung WK, Vasanthan T, Jeon YJ. An anticoagulative polysaccharide from an enzymatic hydrolysate of Ecklonia cava. Carbohyd. Polym. 66: 184-191 (2006)
  13. Kim MM, Ta QV, Mendis E, Rajapakse N, Jung WK, Gyun HG, Jeon YJ, Kim SK. Phlorotannins in Ecklonia cava extract inhibit matrix metalloproteinase activity. Life Sci. 79: 1436-1443 (2006)
  14. Lefkowitz DL, Gelderman MP, Fuhrmann SR, Graham S, Starnes JD, Lefkowitz SS, Bollen A, Moguilevsky N. Neutrophilic lysozymemacrophage interactions perpetuate chronic inflammation associated with experimental arthritis. Clin. Immunol. 91: 145-155 (1999)
  15. Lee JH, Choi SI, Lee YS, Kim GH. Antioxidant and antiinflammatory activities of Allium victorialis subsp. platyphyllum extracts. Food Sci. Biotechnol. 16: 796-801 (2007)
  16. Lee JM, Kim NJ, Cho DH, Chung MY, Hwang KT, Kin HJ, Jun WJ, Park CS. Ethanol extract of Oenanthe javanica modulates inflammatory response by inhibiting NF-$\kappa$B mediated cyclooxygenase-2 expression in RAW 264.7 macrophage. Food Sci. Biotechnol. 15:303-307 (2006)
  17. Harris SG, Padilla J, Koumas L, Ray D, Phipps RP. Prostaglandins as modulators of immunity. Trends Immunol. 23: 144-150 (2002)
  18. MacMicking J, Xie QW, Nathan C. Nitric oxide and macrophage function. Annu. Rev. Immunol. 15: 323-350 (1997)
  19. Kaur G, Hamid H, Ali A, Alam MS, Athar M. Antiinflammatory evaluation of alcoholic extract of galls of Quercus infectoria. J. Ethnopharmacol. 90: 285-292 (2004)
  20. Bourke E, Moynagh PN. Antiinflammatory effects of glucocorticoids in brain cells, independent of NF-kappa B. J. Immunol. 163: 2113-2119 (1999)
  21. Heo SJ, Jeon YJ, Lee J, Kim HT, Lee KW. Antioxidant effect of enzymatic hydrolyzate from a kelp, Ecklonia cava. Algae 18: 341-347 (2003)
  22. Ahn G, Hwang IS, Park EJ, Kim JH, Jeon YJ, Jee Y. Immunomodulatory effects of an enzymatic extract from Ecklonia cava on murine splenocytes. Mar. Biotechnol. 10: 278-289 (2008)
  23. Cumashi A, Ushakova NA, Preobrazhenskaya ME, D'Incecco A, Piccoli A, Totani L, Tinari N, Morozevich GE, Berman AE, Bilan MI, Usov AI, Ustyuzhanina NE, Grachev AA, Sanderson CJ, Kelly M, Rabinovich GA, Iacobelli S, Nifantiev NE, Consorzio Interuniversitario Nazionale per la Bio-Oncologia, Italy. A comparative study of the anti-inflammatory, anticoagulant, antiangiogenic, and antiadhesive activities of nine different fucoidans from brown seaweeds. Glycobiology 17: 541-552 (2007)
  24. Young JM, Young LMD. Pharmacological Methods in the Control of Inflammation. Alan R Liss, New York, NY, USA. p. 259 (2006)
  25. Oh CH, Kang PS, Kim JW, Kwon J, Oh SH. Water extracts of cultured mountain ginseng stimulate immune cells and inhibit cancer cell proliferation. Food Sci. Biotechnol. 15: 369-373 (2006)
  26. Katiyar SK, Mukhtar H. Inhibition of phorbol ester tumor promoter 12-O-tetradecanoylphorbol-13-acetate-caused inflammatory responses in SENCAR mouse skin by black tea polyphenols. Carcinogenesis 18: 1911-1916 (1997)
  27. Sanchez T, Moren JJ. Role of leukocyte influx in tissue prostaglandin H synthase-2 overexpression induced by phorbol ester and arachidonic acid in skin. Biochem. Pharmacol. 58: 877-879 (1999)
  28. Kwan WJ, Han CK, Son KH, Chang HW, Kang SS, Park BK, Kim HP. Effects of ginkgetin from Ginkgo biloba leaves on cyclooxygenases and in vivo skin inflammation, SK Chemicals Ltd. Pharmacology 68: 316-321 (2005)
  29. Park BK, Heo MY, Park H, Kim HP. Inhibition of TPA-induced cyclooxygenase-2 expression and skin inflammation in mice by wogonin, a plant flavone from Scutellaria radix. Eur. J. Pharmacol. 425: 153-157 (2001)
  30. Omata M, Matsui N, Inomata N, Ohno T. Protective effects of polysaccharide fucoidin on myocardial ischemia-reperfusion injury in rats. J. Cardiovasc. Pharm. 30: 717-724 (1997)
  31. Granert C, Raud J, Waage A, Lindquist L. Effects of polysaccharide fucoidin on cerebrospinal fluid interleukin-1 and tumor necrosis factor a in pneumococcal meningitis in the rabbit. Infect. Immun. 67: 2071-2074 (1999)
  32. Foxall C, Watson SR, Dowbenko D, Lasky LA, Kiso M, Hasegawa A, Asa D, Brandley BK. The three members of the selectin receptor family recognize a common carbohydrate epitope, the sialyl Lewis(x) oligosaccharide. J. Cell Biol. 117: 895-902 (1992)
  33. Game SM, Rajapurohit PK, Clifford M, Bird MI, Priest R, Bovin NV, Nifantiev NE, O'Beirne G, Cook ND. Scintillation proximity assay for E-, P-, and L-selectin utilizing polyacrylamide-based neoglycoconjugates as ligands. Anal. Biochem. 258: 127-135 (1998)