Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Anti-inflammatory Activity of an Ethanol Extract of Laminaria japonica Root on Lipopolysaccharide-induced Inflammatory Responses in RAW 264.7 Cells

Lipopolysaccharide로 유도된 RAW 264.7 세포에서 다시마 뿌리 에탄올 추출물의 항염증 효과

  • Kang, Bo-Kyeong (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Kim, Koth-Bong-Woo-Ri (Institute of Fisheries Sciences, Pukyong National University) ;
  • Kim, Min-Ji (Institute of Fisheries Sciences, Pukyong National University) ;
  • Bark, Si-Woo (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Pak, Won-Min (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Kim, Bo-Ram (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Ahn, Na-Kyung (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Choi, Yeon-Uk (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Ahn, Dong-Hyun (Department of Food Science & Technology/Institute of Food Science, Pukyong National University)
  • 강보경 (부경대학교 식품공학과/식품연구소) ;
  • 김꽃봉우리 (부경대학교 수산과학연구소) ;
  • 김민지 (부경대학교 수산과학연구소) ;
  • 박시우 (부경대학교 식품공학과/식품연구소) ;
  • 박원민 (부경대학교 식품공학과/식품연구소) ;
  • 김보람 (부경대학교 식품공학과/식품연구소) ;
  • 안나경 (부경대학교 식품공학과/식품연구소) ;
  • 최연욱 (부경대학교 식품공학과/식품연구소) ;
  • 안동현 (부경대학교 식품공학과/식품연구소)
  • Received : 2014.08.02
  • Accepted : 2014.10.16
  • Published : 2014.12.31
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Abstract

Laminaria japonica roots have not been used practically in Korea. In this study, in order to promote the use of these by-products, the anti-inflammatory activity of an ethanol extract of Laminaria japonica root (LJREE) was investigated using a lipopolysaccharide (LPS) to induce an inflammatory response in RAW 264.7 cells. To examine the potential anti-inflammatory effects of LJREE, levels of nitric oxide (NO) and pro-inflammatory cytokines, such as interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and interleukin-$1{\beta}$ (IL-$1{\beta}$), and cell proliferation were measured. We found that NO levels decreased in a dose-dependent manner, the production of IL-6, TNF-${\alpha}$, and IL-$1{\beta}$ was suppressed, and the production of IL-$1{\beta}$ was inhibited over 30% after treatment with $100{\mu}g/mL$ LJREE. In conclusion, the proliferation of RAW 264.7 cells, measured by MTT assay, confirmed that LJREE may have significant effects on inflammatory factors without any cytotoxicity, making it a potential anti-inflammatory agent.

본 연구에서는 다시마의 가공 시 부산물로 발생하는 다시마 뿌리의 항염증 효과를 알아보기 위하여 LPS로 염증을 유도한 RAW 264.7 세포에 대한 추출물의 효과를 살펴보았다. 먼저 다시마 뿌리 에탄올 추출물이 세포독성을 가지는지 알아보기 위해 MTT assay를 진행한 결과, 모든 첨가 농도에서 대식세포 증식능이 control에 비해 유의적으로 증가하여 세포 독성을 가지지 않는 것을 확인하였다. 세포 독성을 가지지 않았던 농도대(0.1, 1, 10, 50, $100{\mu}g/mL$)에서 염증 억제 효과를 살펴보기 위해 NO 및 염증성 cytokine의 분비 억제 효과를 살펴보았다. 대식세포 배양액 내의 $NO_2{^-}$ 농도를 측정한 결과, 추출물 첨가 농도에 따라 유의적으로 분비량이 감소함을 확인하였으며, 특히 $100{\mu}g/mL$의 농도에서 약 28%의 억제 효과를 보였다. 또한 대식세포의 염증성 cytokine인 IL-6, TNF-${\alpha}$ 및 IL-$1{\beta}$의 분비량을 첨가 농도 의존적으로 억제함을 확인하였다. 특히 IL-$1{\beta}$에 대해, $100{\mu}g/mL$의 농도에서 약 32%의 감소 효과를 보였다. 따라서 다시마 뿌리 에탄올 추출물은 세포 독성을 가지지 않고 염증성 매개 인자인 NO 및 cytokine을 억제하여 항염증 효과를 가짐을 확인하였다.

Keywords

References

  1. Kim YS, Kang CO, Kim MH, Cha WS, Shin HJ. Contents of water extract for Laminaria japonica and its antioxidant activity. Korean Soc. Biotechnol. Bioeng. J. 26: 112-118 (2011) https://doi.org/10.7841/ksbbj.2011.26.2.112
  2. Kang KS, Nam CH, Park EK, Ha BJ. The enzymatic regulatory effects of Laminaria japonica fucoidan extract in hepatotoxicity. Korean J. Life Sci. 16: 1104-1108 (2006) https://doi.org/10.5352/JLS.2006.16.7.1104
  3. Choi JH, Choi JS, Byun DS, Yang DS. Basic studies on the development of diet for the treatment of obesity. II. Comparison of the inhibitory effect of algae and crude drug components on obesity. Korean J. Fish. Aquat. Sci. 19: 485-492 (1986)
  4. Lahaye M. Marine algae as sources of fibres: determination of soluble and insoluble dietary fibre contents in some 'sea vegetables'. J. Sci. Food Agr. 54: 587-594 (1991) https://doi.org/10.1002/jsfa.2740540410
  5. Kim YY, Lee KW, Kim GB, Cho YJ. Studies on physicochemical and biological properties of depolymerized alginate from sea tangle, Laminaria japonicus by thermal decomposition. J. Korean Fish. Soc. 33: 388-392 (2000)
  6. Ryu BH, Kim DS, Cho KJ, Sin DB. Antitumor activity of seaweeds toward sarcoma-180. Korean J. Food Sci. Technol. 21: 595-600 (1989)
  7. Oh CK, Oh MC, Kim SH, Lim SB, Kim SH. Antimutagenic and antimicrobial effect of ethanol extracts from sea-mustard and seatangle. J. Korean Fish. Soc. 31: 90-94 (1998)
  8. Nakashima H, Kido N, Kobayashi N, Motoki M, Neushul M, Yamamoto N. Purification and characterization of an avian myeloblastosis and human immunodeficiency virus reverse transcriptase inhibitor sulfated polysaccharide extracted from sea algae. Antimicrob. Agents Chemother. 31: 1524-1528 (1987) https://doi.org/10.1128/AAC.31.10.1524
  9. Ryu DS, Oh SM, Kim KH, Kim SH, Choi HJ, Lee DS. Immunomodulating activity of Laminaria japonica polysaccharides. Korean J. Food Sci. Technol. 42: 350-354 (2010)
  10. Vishchuk OS, Ermakova SP, Zvyagintseva TN. Sulfated polysaccharides from brown seaweeds Saccharina japonica and Undaria pinnatifida: isolation, structural characteristics, and antitumor activity. Carbohydr. Res. 346: 2769-2776 (2011) https://doi.org/10.1016/j.carres.2011.09.034
  11. Wang J, Zhang Q, Zhang Z, Song H, Li P. Potential antioxidant and anticoagulant capacity of low molecular weight fucoidan fractions extracted from Laminaria japonica. Int. J. Biol. Macromol. 46: 6-12 (2010) https://doi.org/10.1016/j.ijbiomac.2009.10.015
  12. Lee JY, Lee MS, Choi HJ, Choi JW, Shin TS, Woo HC, Kim JI, Kim HR. Hexane fraction from Laminaria japonica exerts antiinflammatory effects on lipopolysaccharide-stimulated RAW 264.7 macrophages via inhibiting NF-kappaB pathway. Eur. J. Nutr. 52: 409-421 (2013) https://doi.org/10.1007/s00394-012-0345-1
  13. Chun JY, Cha SH, Lee JS, Kim YS, Park KP. Extraction of carbohydrates and minerals from laminaria using organic acid. Korean Chem. Eng. Res. 50: 238-243 (2012) https://doi.org/10.9713/kcer.2012.50.2.238
  14. Jung HK, Kang BM, Jang JH, Ahn BK, Yeo JH, Jung WS, Cho JH, Kuk YI, Hyun KH, Cho HW. Inhibitory effect of Alopecurus aequalis sobol ethanol extracts on LPS-induced inflammatory response in RAW 264.7 cells. Korean J. Medicinal Crop Sci. 22: 98-104 (2014) https://doi.org/10.7783/KJMCS.2014.22.2.98
  15. Higuchi M, Hisgahi N, Taki H, Osawa T. Cytolytic mechanism of activated macrophages. Tumor necrosis factor and L-arginine-dependent mechanisms act synergistically as the major cytolytic mechanisms of activated macrophages. J. Immunol. 144: 1425-1431 (1990)
  16. Ljung T, Lundberg S, Varsanyi M, Johansson C, Schmidt PT, Herufl M, Lundberg JO, Hellstrom PM. Rectal nitric oxide as biomarker in the treatment of inflammatory bowel disease: responders versus nonresponders. World J. Gastroenterol. 12: 3368-3392 (2006)
  17. Dogne JM, Hanson J, Supuran C, Pratico D. Coxibs and cardiovascular side-effects: from light to shadow. Curr. Pharm. Design 12: 971-975 (2006) https://doi.org/10.2174/138161206776055949
  18. Makins R, Ballinger A. Gastrointestinal side effects of drugs. Expert Opin. Drug Saf. 2: 421-429 (2003) https://doi.org/10.1517/14740338.2.4.421
  19. Kim YJ, Son DY. Inflammatory mediator regulation of the Zizyphus jujube leaf fractions in the LPS-stimulated Raw 264.7 mouse macrophage. Korean J. Food Preserv. 21: 114-120 (2014) https://doi.org/10.11002/kjfp.2014.21.1.114
  20. Leem HH, Kim EO, Seo MJ, Choi SW. Antioxidant and anti-inflammatory activities of eugenol and its derivatives from clove (Eugenia caryophyllata Thunb.). J. Korean Soc. Food Sci. Nutr. 40: 1361-1370 (2011) https://doi.org/10.3746/jkfn.2011.40.10.1361
  21. Park YM, Won JH, Yun KJ, Ryu JH, Han YN, Choi SK, Lee KT. Preventive effect of Ginkgo biloba extract (GBB) on the lipopolysaccharide-induced expressions of inducible nitric oxide synthase and cyclooxygenase-2 via suppression nuclear factor-${\kappa}B$ in RAW 264.7 cells. Biol. Pharm. Bull. 29: 985-990 (2006) https://doi.org/10.1248/bpb.29.985
  22. Lee ST, Jeong YR, Ha MH, Kim SH, Byun MW, Jo SK. Induction of nitric oxide and TNF-${\alpha}$ by herbal plant extract in mouse macrophage. J. Korean Soc. Food Sci. Nutr. 29: 342-348 (2000)
  23. Lee YH, Lim EM. Anti-inflammatory effect of Ligustri lucidi fructus water extract in RAW 264.7 cells induced by LPS. J. Korean Obstet. Gynecol. 26: 66-81 (2013)
  24. Yoon SB, Han HS, Lee YJ. Effect of Scutellariae radix extract on the proinflammatory mediators in RAW 264.7 cells induced by LPS. Korean J. Herbology 26: 75-81 (2011)
  25. Weisz A, Cicatiello L, Esumi H. Regulation of the mouse inducible-type nitric oxide synthase gene promoter by interferongamma, bacterial lipopolysaccharide and NG-monomethyl-L-arginine. Biochem. J. 316: 209-215 (1996)
  26. An SM, Kim HG, Choi EJ, Hwang HH, Lee ES, Baek JH, Boo YC, Koh JS. Screening for anti-inflammatory activities in extracts from Korean herb medicines. J. Soc. Cosmet. Scientists Korea 40: 95-108 (2014) https://doi.org/10.15230/SCSK.2014.40.1.95
  27. Kang CH, Choi YH, Choi IW, Lee JD, Kim GY. Inhibition of lipopolysaccharide-induced iNOS, COX-2, and TNF-${\alpha}$ expression by aqueous extract of Orixa Japonica in RAW 264.7 cells via suppression of NF-${\kappa}B$ activity. Trop. J. Pharm. Res. 10: 161-168 (2011)
  28. Miyataka M, Rich KA, Ingram M, Yamamoto T, Bing RJ. Nitric oxide, anti-inflammatory drugs on renal prostaglandins and cyclooxygenase-2. Hypertension 39: 785-789 (2002) https://doi.org/10.1161/hy0302.105689
  29. Choi JS, Shin SH, Ha YM, Kim YC, Kim TB, Park SM, Choi IS, Song HJ, Choi YJ. Mineral contents and physiological activities of dried sea tangle (Laminaria japonica) collected from Gijang and Wando in Korea. Korean J. Life Sci. 18: 474-481 (2008) https://doi.org/10.5352/JLS.2008.18.4.474
  30. Adams JL, Czuprynski CJ. Mycobacterial cell wall components induce the production of TNF-${\alpha}$, IL-1, and IL-6 by bovine monocytes and the murine macrophage cell line RAW 264.7. Microb. Pathogenesis 16: 401-411 (1994) https://doi.org/10.1006/mpat.1994.1040
  31. Iwamoto M, Kurachi M, Nakashima T, Kim DK, Yamaguchi K, Oda T, Iwamoto Y, Muramatsu T. Structure-activity relationship of alginate oligosaccharides in the induction of cytokine production from RAW 264.7 cells. FEBS Lett. 579: 4423-4429 (2005) https://doi.org/10.1016/j.febslet.2005.07.007
  32. Choi SB, Bae GS, Jo IJ, Park KC, Seo SH, Kim DG, Shin JY, Gwak TS, Lee JH, Lee GS, Park SJ, Song HJ. The anti-inflammatory effect of Lithospermum Erythrorhizon on lipopolysaccharide-induced inflammatory response in RAW 264.7 cells. Korean J. Herbology 28: 67-73 (2013) https://doi.org/10.6116/kjh.2013.28.2.67
  33. Bhattacharyya A, Pathak S, Datta S, Chattopadhyay S, Basu J, Kundu M. Mitogen-activated protein kinases and nuclear factorkappaB regulate helicobacter pylori-mediated interleukin-8 release from macrophages. Biochem. J. 368: 121-129 (2002) https://doi.org/10.1042/BJ20020555
  34. Kim BW, Kim JI, Kim HR, Byun DS. Anti-inflammatory effect of an ethyl acetate fraction from Myagropsis yendoi on lipopolysaccharides-stimulated RAW 264.7 cells. Korean J. Fish. Aquat. Sci. 47: 527-536 (2014) https://doi.org/10.5657/KFAS.2014.0527
  35. Hibi M, Nakajima T, Hirano T. IL-6 cytokine family and signal transduction: a model of the cytokine system. J. Mol. Med. 74: 1-12 (1996) https://doi.org/10.1007/BF00202068
  36. Kim HJ, Park TS, Jung MS, Son JH. Study on the anti-oxidant and anti-inflammatory activities of sarcocarp and calyx of persimmon (Cheongdo Bansi). J. Appl. Biol. Chem. 54: 71-78 (2011) https://doi.org/10.3839/jabc.2011.013
  37. Wang X, Luo Y, Liao WB, Zhang J, Chen TM. Effect of osteoprotegerin in combination with interleukin-6 on inhibition of osteoclast differentiation. Chin. J. Traumatol. 16: 277-280 (2013)
  38. Kim YJ, Son DY. Antioxidant and inhibitory effects of korean Panax ginseng extract on pro-inflammatory mediators in LPS-induced RAW 264.7 macrophages. J. Korean Soc. Food Sci. Nutr. 41: 1371-1377 (2012) https://doi.org/10.3746/jkfn.2012.41.10.1371
  39. Park SG, Jegal KH, Jung JY, Back YD, Byun SH, Kim YW, Cho IJ, Park SM, Kim SC. Leonuri fructus ameliorates acute inflammation via the inhibition of NF-${\kappa}B$-mediated nitric oxide and pro-inflammatory cytokine production. Korean J. Orient. Physiol. Pathol. 28: 178-185 (2014)
  40. Moon JH, Go H, Shin SM, Kim KT. Anti-inflammatory effect of extracts from Ligustrum obtusifolium S. fruits in RAW 264.7 macrophages. J. Soc. Korean Med. Diagn. 17: 263-273 (2013)
  41. Tracey D, Klareskog L, Sasso EH, Salfeld JG, Tak PP. Tumor necrosis factor antagonist mechanisms of action: a comprehensive review. Pharmacol. Ther. 117: 244-279 (2008) https://doi.org/10.1016/j.pharmthera.2007.10.001
  42. Giuliani C, Napolitano G, Bucci I, Montani V, Monaco F. NF-${\kappa}B$ transcription factor: role in the pathogenesis of inflammatory, autoimmune, and neoplastic diseases and therapy implications. Clin. Ter. 152: 249-253 (2001)
  43. Medzhitov R. Origin and physiological roles of inflammation. Nature 454: 428-435 (2008) https://doi.org/10.1038/nature07201
  44. Tak PP, Firestein GS. NF-${\kappa}B$: a key role in inflammatory diseases. J. Clin. Invest. 107: 7-11 (2001) https://doi.org/10.1172/JCI11830
  45. Lee AK, Sung SH, Kim YC, Kim SG. Inhibition of lipopolysaccharide-inducible nitric oxide synthase, TNF-${\alpha}$ and COX-2 expression by sauchinone effects on I-${\kappa}B{\alpha}$ phosphorylation, C/EBP and AP-1 activation. Br. J. Pharmacol. 139: 11-20 (2003) https://doi.org/10.1038/sj.bjp.0705231
  46. Delgado AV, McManus AT, Chambers JP. Production of tumor necrosis factor-alpha, interleukin 1-beta, interleukin 2, and interleukin 6 by rat leukocyte subpopulations after exposure to substance P. Neuropeptides 37: 355-361 (2003) https://doi.org/10.1016/j.npep.2003.09.005
  47. Masters SL, Simon A, Aksentijevich I, Kastner DL. Horror autoinflammaticus: the molecular pathophysiology of autoinflammatory disease. Ann. Rev. Immunol. 27: 621-668 (2009) https://doi.org/10.1146/annurev.immunol.25.022106.141627

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