Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Anti-Inflammatory Effect of Ethanol Extract from Grateloupia elliptica Holmes on Lipopolysaccharide-Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears

LPS로 유도된 RAW 264.7 세포와 마우스 귀 조직에 대한 참도박(Grateloupia elliptica Holmes) 에탄올 추출물의 항염증 효과

  • Bae, Nan-Young (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Kim, Min-Ji (Institute of Fisheries Sciences, Pukyong National University) ;
  • Kim, Koth-Bong-Woo-Ri (Institute of Fisheries Sciences, 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) ;
  • Park, Ji-Hye (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Park, Sun-Hee (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)
  • 배난영 (부경대학교 식품공학과/식품연구소) ;
  • 김민지 (부경대학교 수산과학연구소) ;
  • 김꽃봉우리 (부경대학교 수산과학연구소) ;
  • 안나경 (부경대학교 식품공학과/식품연구소) ;
  • 최연욱 (부경대학교 식품공학과/식품연구소) ;
  • 박지혜 (부경대학교 식품공학과/식품연구소) ;
  • 박선희 (부경대학교 식품공학과/식품연구소) ;
  • 안동현 (부경대학교 식품공학과/식품연구소)
  • Received : 2015.06.09
  • Accepted : 2015.07.22
  • Published : 2015.08.31
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Abstract

This study investigated the anti-inflammatory effects of ethanol extract from Grateloupia elliptica Holmes (GEHEE) on the lipopolysaccharide-induced inflammatory response. Anti-inflammatory effects were detected by enzyme-linked immunosorbent assay, Western blotting, and immunohistochemistry. There were no cytotoxic effects on proliferation of macrophages treated with GEHEE compared to the control. GEHEE remarkably suppressed NO and pro-inflammatory cytokines (interleukin-6, tumor necrosis $factor-{\alpha}$, and $interleukin-1{\beta}$) production and reduced expression of inducible nitric oxide synthase, cyclooxygenase-2, and nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) proteins in a dose-dependent manner. GEHEE also significantly reduced activation of mitogen-activated protein kinases (MAPKs). The formation of edema in mouse ears was reduced at the highest dose compared to the control. GEHEE also reduced dermal thickness and mast cell numbers based on histological analysis. These results suggest that GEHEE exerts significant anti-inflammatory activity via inhibition of $NF-{\kappa}B$ and MAPKs activation and may be a potential anti-inflammatory therapeutic material.

본 연구에서는 참도박 에탄올 추출물(GEHEE)의 항염증 효과를 알아보기 위해 lipopolysaccharide에 의해 활성화된 대식세포로부터 분비되는 염증매개인자들의 발현량과 마우스 모델을 이용한 귀 부종 및 조직학적 관찰 실험을 진행하였다. 그 결과 염증을 유발하는 대표적 물질인 NO 및 사이토카인[interleukin(IL)-6, $IL-1{\beta}$ 및 tumor necrosis factor $receptor-{\alpha}$]의 분비량이 GEHEE 농도 의존적으로 유의적 감소를 보였다. 또한 전사인자인 nuclear $factor-{\kappa}B$의 활성과 인산화효소인 mitogen-activated protein kinases(p38, JNK 및 ERK)의 발현량이 억제됨을 확인함에 따라 염증작용 기전에서 이들의 활성 억제가 NO 및 사이토카인 분비량 조절에 영향을 미침을 확인하였다. 동물모델을 이용한 실험에서는 croton oil로 부종을 유발한 마우스 귀 조직에 GEHEE를 처리하였을 때 항염증제인 prednisolone 처리구와 유사한 수준으로 경피 및 진피의 두께가 감소한 것을 확인하였으며, 조직 내 침윤되는 mast cell의 수도 현저히 억제됨을 확인하였다. 따라서 본 연구 결과는 참도박 에탄올 추출물의 항염증 효능을 가진 기능성 소재로의 이용 가능성을 제시한다.

Keywords

References

  1. Willoughby DA. 1975. Human arthritis applied to animal models. Towards a better therapy. Ann Rheum Dis 34: 471-478. https://doi.org/10.1136/ard.34.6.471
  2. Guha M, 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
  3. Miyake K. 2004. Innate recognition of lipopolysaccharide by Toll-like receptor 4-MD-2. Trends Microbiol 12: 186-192. https://doi.org/10.1016/j.tim.2004.02.009
  4. Hofseth LJ, Ying L. 2006. Identifying and defusing weapons of mass inflammation in carcinogenesis. Biochim Biophys Acta 1765: 74-84.
  5. Kim YS, Lee SJ, Hwang JW, Kim EH, Park PJ, Jeong JH. 2012. Anti-inflammatory effects of extracts from Ligustrum ovalifolium H. leaves on RAW 264.7 macrophages. J Korean Soc Food Sci Nutr 41: 1205-1210. https://doi.org/10.3746/jkfn.2012.41.9.1205
  6. Lee JY, Ahn EK, Ko HJ, Cho YR, Ko W, Jung YH, Choi KM, Choi MR, Oh JS. 2014. Anti-melanogenic, anti-wrinkle, anti-inflammatory and anti-oxidant effects of Xylosma congesta leaf ethanol extract. J Appl Biol Chem 57: 365-371. https://doi.org/10.3839/jabc.2014.058
  7. Kim ES, Jeong NH. 2014. Anti-inflammatory effect of germinated mung bean and hairdye applications. J Korean Oil Chemists' Soc 31: 23-30. https://doi.org/10.12925/jkocs.2014.31.1.23
  8. Jung YS, Eun CS, Jung YT, Kim HJ, Yu MH. 2013. Antiinflammatory effects of Picrasma quassioides (D.DON) BENN leaves extract. J Life Sci 23: 629-636. https://doi.org/10.5352/JLS.2013.23.5.629
  9. Park JC, Jang YI, Doo MS, Kim SH, Choi JW. 1996. Effect of methanolic extract of Pachymeniopsis elliptica on lipids component of hyperlipidemic rats. J Korean Soc Food Sci Nutr 25: 958-962.
  10. Yoon JA, Yu KW, Jun WJ, Cho HY, Son YS, Yang HC. 2000. Purification of blood anticoagulant polysaccharide from Pachymeniopsis elliptica. J Korean Soc Food Sci Nutr 29: 908-916.
  11. Lee SY. 2002. Studies on the antioxidants from the Phelinus linteus and Grateloupia elliptica. MS Thesis. Hanbat National University, Daejeon, Korea.
  12. Kang JI, Kim SC, Han SC, Hong HJ, Jeon YJ, Kim B, Ko YS, Yoo ES, Kang HK. 2012. Hair-loss preventing effect of Grateloupia elliptica. Biomol Ther 20: 118-124. https://doi.org/10.4062/biomolther.2012.20.1.118
  13. Park YM, Won JH, Yun KJ, Ryu JH, Han YN, Choi SK, Lee KT. 2006. Preventive effect of Ginkgo biloba extract (GBB) on the lipopolysaccharide-induced expressions of inducible nitric oxide synthase and cyclooxygenase-2 via suppression of nuclear factor-${\kappa}B$ in RAW 264.7 cells. Biol Pharm Bull 29: 985-990. https://doi.org/10.1248/bpb.29.985
  14. Lee ST, Jeong YR, Ha MH, Kim SH, Byun MW, Jo SK. 2000. Induction of nitric oxide and TNF-α by herbal plant extracts in mouse macrophages. J Korean Soc Food Sci Nutr 29: 342-348.
  15. Sheeba MS, Asha VV. 2009. Cardiospermum halicacabum ethanol extract inhibits LPS induced COX-2, TNF-${\alpha}B$ and iNOS expression, which is mediated by NF-${\kappa}B$ regulation, in RAW264.7 cells. J Ethnopharmacol 124: 39-44. https://doi.org/10.1016/j.jep.2009.04.020
  16. Laemmli UK. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685. https://doi.org/10.1038/227680a0
  17. Towbin H, Staehelin T, Gordon J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350-4354. https://doi.org/10.1073/pnas.76.9.4350
  18. Kim DW, Chi YS, Son KH, Chang HW, Kim JS, Kang SS, Kim HP. 2002. Effects of sophoraflavanone G, a prenylated flavonoid from Sophora flavescens, on cyclooxygenase-2 and in vivo inflammatory response. Arch Pharm Res 25:329-335. https://doi.org/10.1007/BF02976635
  19. Saraiva RA, Araruna MK, Oliveira RC, Menezes KD, Leite GO, Kerntopf MR, Costa JG, Rocha JB, Tomé AR, Campos AR, Menezes IR. 2011. Topical anti-inflammatory effect of Caryocar coriaceum Wittm. (Caryocaraceae) fruit pulp fixed oil on mice ear edema induced by different irritant agents. J Ethnopharmacol 136: 504-510. https://doi.org/10.1016/j.jep.2010.07.002
  20. Lee HN, Kim JK, Kwon GT, Shim JH, Kim JD, Yoon Park JH. 2012. Anti-inflammatory effects of ethanol extract from bark of Acer barbinerve maxim. J Korean Soc Food Sci Nutr 41: 1242-1247. https://doi.org/10.3746/jkfn.2012.41.9.1242
  21. Jeong H, Sung M, Kim Y, Ham H, Choi Y, Lee J. 2012. Anti-inflammatory activity of Salvia plebeia R. Br. leaf through heme oxygenase-1 induction in LPS-stimulated RAW264.7 macrophages. J Korean Soc Food Sci Nutr 41:888-894. https://doi.org/10.3746/jkfn.2012.41.7.888
  22. Kim JY, Jung KS, Jeong HG. 2004. Suppressive effects of the kahweol and cafestol on cyclooxygenase-2 expression in macrophages. FEBS Lett 569: 321-326. https://doi.org/10.1016/j.febslet.2004.05.070
  23. Jeong DH, Kim KBWR, Kang BK, Jung SA, Kim HJ, Jeong HY, Bark SW, Ahn DH. 2012. Anti-inflammatory activity of the Undaria pinnatifida water extract. J Appl Biol Chem 55: 221-225. https://doi.org/10.3839/jabc.2012.035
  24. Jeong DH, Kang BK, Kim KBWR, Kim MJ, Ahn DH. 2014. Anti-inflammatory activity of Sargassum micracanthum water extract. J Appl Biol Chem 57: 227-234. https://doi.org/10.3839/jabc.2014.036
  25. Hyun TK, Ko YJ, Kim EH, Chung IM, Kim JS. 2015. Antiinflammatory activity and phenolic composition of Dendropanax morbifera leaf extracts. Ind Crop Prod 74: 263-270. https://doi.org/10.1016/j.indcrop.2015.05.002
  26. Chen X, Miao J, Wang H, Zhao F, Hu J, Gao P, Wang Y, Zhang L, Yan M. 2015. The anti-inflammatory activities of Ainsliaea fragrans Champ. extract and its components in lipopolysaccharide-stimulated RAW 264.7 macrophages through inhibition of NF-${\kappa}B$ pathway. J Ethnopharmacol 170: 72-80. https://doi.org/10.1016/j.jep.2015.05.004
  27. Al-Banna N, Raghupathy R, Albert MJ. 2008. Correlation of proinflammatory and anti-inflammatory cytokine levels with histopathological changes in an adult mouse lung model of Campylobacter jejuni infection. Clin Vaccine Immunol 15: 1780-1787. https://doi.org/10.1128/CVI.00193-08
  28. Fernando MR, Reyes JL, Iannuzzi J, Leung G, McKay DM. 2014. The pro-inflammatory cytokine, interleukin-6, enhances the polarization of alternatively activated macrophages. PLoS ONE 9: e94188. https://doi.org/10.1371/journal.pone.0094188
  29. Jeong DH, Kim KBWR, Kim MJ, Kang BK, Bark SW, Pak WM, Kim BR, Ahn NK, Choi YU, Ahn DH. 2014. Anti-inflammatory effect of ethanol extract from Sargassum fulvellum on lipopolysaccharide induced inflammatory responses in RAW 264.7 cells and mice ears. J Korean Soc Food Sci Nutr 43: 1158-1165. https://doi.org/10.3746/jkfn.2014.43.8.1158
  30. Li YY, Huang SS, Lee MM, Deng JS, Huang GJ. 2015. Anti-inflammatory activities of cardamonin from Alpinia katsumadai through heme oxygenase-1 induction and inhibition of NF-${\kappa}B$ and MAPK signaling pathway in the carrageenan-induced paw edema. Int Immunopharmacol 25:332-339. https://doi.org/10.1016/j.intimp.2015.02.002
  31. Miyasaka N, Hirata Y. 1997. Nitric oxide and inflammatory arthritides. Life Sci 61: 2073-2081. https://doi.org/10.1016/S0024-3205(97)00585-7
  32. Simmons ML, Murphy S. 1992. Induction of nitric oxide from glial cells. J Neurochem 59: 897-905. https://doi.org/10.1111/j.1471-4159.1992.tb08328.x
  33. Martel-Pelletier J, Pelletier JP, Fahmi H. 2003. Cyclooxygenase-2 and prostaglandins in articular tissues. Semin Arthritis Rheum 33: 155-167. https://doi.org/10.1016/S0049-0172(03)00134-3
  34. Needleman P, Isakson PC. 1997. The discovery and function of COX-2. J Rheumatol Suppl 49: 6-8.
  35. Seybold VS, Jia YP, Abrahams LG. 2003. Cyclo-oxygenase- 2 contributes to central sensitization in rats with peripheral inflammation. Pain 105: 47-55. https://doi.org/10.1016/S0304-3959(03)00254-9
  36. Xie QW, Whisnant R, 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
  37. Kim B, Kim JI, Kim HR, Byun DS. 2014. Anti-inflammatory effect of an ethyl acetate fraction from Myagropsis yendoi on lipopolysaccharides-stimulated RAW 264.7 cells. Kor J Fish Aquant Sci 47: 527-536.
  38. Dong C, Davis RJ, Flavell RA. 2002. MAP kinases in the immune response. Annu Rev Immunol 20: 55-72. https://doi.org/10.1146/annurev.immunol.20.091301.131133
  39. Kim HG, Shrestha B, Lim SY, Yoon DH, Chang WC, Shin DJ, Han SK, Park SM, Park JH, Park HI, Sung JM, Jang Y, Chung N, Hwang KC, Kim TW. 2006. Cordycepin inhibits lipopolysaccharide-induced inflammation by the suppression of NF-${\kappa}B$ through Akt and p38 inhibition in RAW 264.7 macrophage cells. Eur J Pharmacol 545: 192-199. https://doi.org/10.1016/j.ejphar.2006.06.047
  40. Park JS, Lee EJ, Lee JC, Kim WK, Kim HS. 2007. Anti-inflammatory effects of short chain fatty acids in IFN-${\gamma}$-stimulated RAW 264.7 murine macrophage cells: Involvement of NF-${\kappa}B$ and ERK signaling pathways. Int Immunopharmacol 7: 70-77. https://doi.org/10.1016/j.intimp.2006.08.015
  41. Towbin H, Pignat W, Wiesenberg I. 1995. Time-dependent cytokine production in the croton oil-induced mouse ear oedema and inhibition by prednisolone. Inflamm Res 44: S160-161. https://doi.org/10.1007/BF01778311
  42. Kang BK, Kim KBWR, Kim MJ, Bark SW, Pak WM, Kim BR, Ahn NK, Choi YU, Ahn DH. 2014. Anti-inflammatory activity of an ethanol extract of Laminaria japonica root on lipopolysaccharide-induced inflammatory response in RAW 264.7 cells. Korean J Food Sci Technol 46: 729-733. https://doi.org/10.9721/KJFST.2014.46.6.729
  43. Jeong DH, Kim KB, Kim MJ, Kang BK, Ahn DH. 2014. Anti-inflammatory activity of methanol extract and n-hexane fraction mojabanchromanol b from Myagropsis myagroides. Life Sci 114: 12-19. https://doi.org/10.1016/j.lfs.2014.07.036

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