Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
권호 표지
DOI QR코드

DOI QR Code

Anti-inflammatory Activity of Sargassum micracanthum Water Extract

잔가시 물 추출물의 항염증 효과

  • Jeong, Da Hyun (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • 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) ;
  • Ahn, Dong Hyun (Department of Food Science & Technology/Institute of Food Science, Pukyong National University)
  • Received : 2013.10.16
  • Accepted : 2014.02.09
  • Published : 2014.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

The anti-inflammatory effect of Sargassum micracanthum water extract (SMWE) was investigated using lipopolysaccharide (LPS)-induced inflammatory response in this study. The murine macrophage cell line RAW 264.7 cells were used and MTT assay was performed to measure the cell proliferation ability. The secretion of nitric oxide (NO), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-6 (IL-6), and IL-$1{\beta}$ was measured in LPS-induced RAW 264.7 cells by ELISA. The expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and nuclear transcription factor-kappa B p65 protein was studied by immunoblotting. The Balb/c mice were used for an acute toxicity test, and imprinting control region mice were purchased to evaluate a croton oil-induced ear edema. As a result, there was no cytotoxicity in the macrophage proliferation treated with SMWE compared to the control. NO levels decreased with increasing concentration of SMWE and were inhibited over 50%. Moreover, the secretion of IL-6, TNF-${\alpha}$, and IL-$1{\beta}$ was suppressed in a dose-dependent manner, especially, IL-$1{\beta}$ inhibition activity was over 50% at 50 ${mu}g$/mL. The formation of ear edema of mice was reduced at the highest dose tested compared to that in the control. Moreover, in acute toxicity test, no moralities occurred in mice administered 5,000 mg/kg body weight of SMWE over 2 weeks observation period. These results suggested that SMWE may have significant effects on inflammatory factors and be potential anti-inflammatory therapeutic materials.

본 연구에서는 LPS로 유도된 RAW 264.7 대식세포의 염증반응을 통해 잔가시 모자반 (S. micracanthum) 물 추출물의 항염증 활성을 알아보았다. 잔가시 모자반 물 추출물이 대식세포에 미치는 독성을 알아보기 위해 MTT assay를 시행했으며, NO 생성량을 비롯하여 TNF-${\alpha}$, IL-6 및 IL-$1{\beta}$와 같은 염증 매개성 사이토카인 분비량을 측정하기 위해 ELISA법을 사용하였다. 또한, immunoblotting을 통해 iNOS, COX-2 및 NF-${\kappa}B$ p65 단백질 발현량을 알아보았다. 실험결과, 잔가시 모자반 물 추출물이 대식세포에 미치는 독성은 보이지 않았으며, NO 및 염증 매개성 사이토카인의 분비량이 농도 의존적으로 억제됨을 보였다. 특히, IL-$1{\beta}$ 분비량이 $50{\mu}g/mL$에서 50% 이상 저해됨을 보였다. 또한, iNOS, COX-2 및 NF-${\kappa}B$ p65 단백질 발현량에서도 농도의존적인 감소를 보였다. 모자반 물 추출물을 5,000 mg/kg body weight까지 경구투여 한 후 2주 동안 관찰한 결과 경구독성을 보이지 않음을 확인하였다. 그 후, 동물실험을 통해 염증으로 인한 귀 부종의 두께를 측정한 결과, 가장 높은 처리 농도인 250 mg/kg body weight으로 경구투여 하였을 때 prednisolone을 투여한 대조군과 유의적으로 비슷한 수준까지 귀 부종이 완화됨을 보였다. 따라서 본 연구는 잔가시 모자반 물 추출물이 뛰어난 항염증 효과를 가지고 있음을 나타내며, 향후 염증질환 치료제 개발에 잔가시 모자반 물 추출물이 이용될 수 있을 것으로 사료된다.

Keywords

References

  1. Baek YM, Choi JY, Lee CW, Jeon YS, Han JT, Jang SI et al. (2012) Effects of Chinemys reevesii on lipopolysaccharide-induced inflammatory reactions. Korean J Orinet Physiol Pathol 26, 26-34.
  2. Beutler B and Cerami A (1989) The biology of cachectin/TNF-$\alpha$ primary mediator of the host response. Annu Rev Immunol 7, 625-55. https://doi.org/10.1146/annurev.iy.07.040189.003205
  3. Bishop-Bailey D, Calatayud S, Warner TD, Hla T, and Mitchell JA (2002) Prostaglandins and the regulation of tumor growth. J Environ Pathol Toxicol Oncol 21, 93-101.
  4. Celec P (2004) Nuclear factor kappa B-molecular biomedicine: the next generation. Biomed Pharmacother 58, 365-71. https://doi.org/10.1016/j.biopha.2003.12.015
  5. Chae SY, Kim MJ, Kim DS, Park JE, Jo SK, and Yee ST (2007) Effect of Asterina pectimifera extraction on the activation of immune cells. J Korean Soc Food Sci Nutr 36, 269-375. https://doi.org/10.3746/jkfn.2007.36.3.269
  6. Cho EK and Choi YJ (2010) Physiological activities of hot water extracts from Ecklonia cava Kjellman. J Life Sci 20, 1675-82. https://doi.org/10.5352/JLS.2010.20.11.1675
  7. Delgado AV, McManus AT, and Chambers JP (2003) Production of tumor necrosis factor-alpha, interleukin 1-beta, interleukin 2 and interleukin 6 by rat leukocyte subpopulations after exposure to substance P. Neuro 37, 355-61.
  8. Ghosh S and Hayden HS (2008) New regulators of NF-$\kappa{B}$ in inflammation. Nat Rev Immunol. 8, 837-48. https://doi.org/10.1038/nri2423
  9. Ham YM, Kim KN, Lee WJ, Lee NH, and Hyun CG (2010) Chemical constituents from Sargassum micracanthum and antioxidant activity. Int J Pharmacol 6, 147-51. https://doi.org/10.3923/ijp.2010.147.151
  10. Heinzemann A and Daser A (2002) Mouse models for the genetic dissection of atopy. Int Arch Allergy Immunol 127, 170-80. https://doi.org/10.1159/000053861
  11. Higuchi M, Higashi N, Taki H, and Osawa T (1990) Cytolytic mechanism of activated macrophages. Tumor necrosis factor and L-arginine-dependent mechanism acts as synergistically as the major cytolytic mechanisms of activated macrophages. J Immunol 144, 1425-31.
  12. Hume DA, Wells CA, and Ravasi T (2007) Transcriptional regulatory networks in macrophages. Novartis Found Symp 281, 2-18.
  13. Imm JY and Kim SJ (2010) Anti-cancer and ant-inflammatory effects of mung bean and soybean extracts. Korean J Food Sci Technol 42, 755-61.
  14. Ju MS, Jeong HU, Kim HG, Park GH, Youn YS, Kim YO et al. (2010) Antinociceptive and anti-inflammatory effects of Geranii herba. Korea J Herbol 25, 97-101.
  15. Kim BH, Oh JM, Kang KW, Kwak SH, Yun SY, Lee CH et al. (2008) Evaluation of oxy-radical scavenging capacity of fucoidan. J Environ Toxicol 23, 41-5.
  16. Kim C, Lee IK, Cho GY, Oh KH, Lim YW, and Yun BS (2012a) Sargassumol, a novel antioxidant from the brown alga Sargassum micracanthum. J Antibiot 65, 87-9. https://doi.org/10.1038/ja.2011.107
  17. Kim DH, Hwang EY, and Son JH (2013) Anti-inflammatory activity of Carthamus tinctorious seed extracts in RAW 264.7 cells. J Life Sci 23, 55-62. https://doi.org/10.5352/JLS.2013.23.1.55
  18. Kim DH, Park SJ, Jung JY, Kim SC, and Byun SH (2009) Anti-inflammatory effects of the aqueous extract of Hwangnyeonhaedok-tang in LPSactivated macrophage cells. Kor J Herbol 24, 39-47.
  19. Kim JY, Jung KS, and Jeong HG (2004) Suppressive effects of the kahweol and cafestol on cyclooxygenase-2 expression in macrophages. FEBS Lett 569, 321-6. https://doi.org/10.1016/j.febslet.2004.05.070
  20. Kim MJ (2012) Anti-inflammatory activity of the Sargassum fulvellum and Sargassum sagamianum ethanol extracts. MS Thesis, Pukyong national university, Korea.
  21. Kim NY, Kim HJ, Lee JH, Lee EK, Kang OH, So HS et al. (2011a) Comparison of anti-inflammatory effects of water fermented and ethanol fermented extracts from Rhei Radix et Rhizoma. Korean J Oreint Physiol Pathol 25, 227-33.
  22. Kim RG, Shin KM, Chun SK, Ji SY, Seo SH, Park HJ et al. (2002) In vitro antiinflammatory activity of the essential oil from Ligularia fischeri var. spiciformis in murine macrophage RAW 264.7 cells. Yakhak Hoeji 46, 343-7.
  23. Kim S, Jung E, Shin S, Kim M, Kim YS, Lee J et al. (2011b) Anti-inflammatory activity of Camellia japonica oil. BMB Rep 45, 177-82.
  24. Kim YS, Lee SJ, Hwang JW, Kim EH, Park PJ, and Jeong JH (2012b) Antiinflammatory effects of extracts from Ligustrum ovalifolium H. leaves on RAW 264.7 macrophages. J Korean Soc Food Sci Nutr 41, 1205-10. https://doi.org/10.3746/jkfn.2012.41.9.1205
  25. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-5. https://doi.org/10.1038/227680a0
  26. Lee ES, Ku HK, Moon TC, Lee E, Ahng Y, Lee SH et al. (2004) Inhibition of nitric oxide and tumor necrosis factor-$\alpha$ (TNF-$\alpha$) production by propenone compound through blockade of nuclear factor (NF)-kappa B activation in cultured murine macrophages. Bio Pharm Bull 27, 617-20. https://doi.org/10.1248/bpb.27.617
  27. Lee HN, Kim JK, Kwon GT, Shim JH, Kim JD, and Yoon JH (2012a) Antiinflammatory effects of ethanol extract from Bark of Acer barbinerve Maxim. J Korean Soc Food Sci Nutr 41, 1242-7. https://doi.org/10.3746/jkfn.2012.41.9.1242
  28. Lee KH, Nam HO, and Yoon WH (2007) Effect of protein-bond polysaccharide isolated from Acanthopanax senthopanax in reducing the toxic effect of cisplatin. Korean J Pharmacogn 38, 1-17.
  29. Lee SE, Lee H, Kim K, Kim GS, Kim YO, Soe S et al. (2011) Antiinflammatory activity of medicinal plant extracts. Korean J Medicinal Crop Sci 19, 217-26. https://doi.org/10.7783/KJMCS.2011.19.4.217
  30. Lee ST, Jeong YR, Ha MH, Kim SH, and Byun MW (2000) Induction of nitric oxide and TNF-$\alpha$ by herbal plant extract in mouse macrophage. J Korean Soc Food Sci Nutr 29, 342-8.
  31. Lee YJ, Yoon BR, Choi HS, Lee BY, and Lee OH (2012b) Effect of Sargassum micracanthum extract on lipid accumulation and reactive oxygen species (ROS) production during differentiation of 3T3-L1 preadipocytes. Korean J Food Preserv 19, 455-61. https://doi.org/10.11002/kjfp.2012.19.3.455
  32. Lim H and Shin S (2010) Effects of the essential oil components from Ligusticum chuanxiong on proinflammtory mediators of RAW 264.7 macrophage cells. Nat Prod Sci16, 259-64.
  33. Majdalawieh A and Ro HS (2010) Regulation of $I\kappa{B}\alpha$ function and NF-$\kappa{B}$ signaling: AEBP1 is a novel proinflammatory mediator in macrophages. Mediat Inflamm 2010, 1-27.
  34. Marwaha V, YH Chen, E Helms, S Arad, H Inoue, E Bord et al. (2005) Toligo treatment decreases constitute and UVB-induced COX-2 promoter. J Biol Chem 280, 32379-88. https://doi.org/10.1074/jbc.M503245200
  35. Mathiak G, Grass G, Herzmann T, and Luebke T (2000) Capase-1-inhibitor ac-YVAD-cmk reduces LPS-lethality in rats without affecting haematology or cytokine responses. Br J Pharmacol 131, 383-6. https://doi.org/10.1038/sj.bjp.0703629
  36. McDaniel ML, Kwon G, Hill JR, Marshall CA, and Corbett JA (1996) Cytokines and nitric oxide in islet inflammation and diabetes. Proc Soc Exp Biol Med 211, 24-32. https://doi.org/10.3181/00379727-211-43950D
  37. Sanchez-Borges M (2010) NSAID hypersensitivity (respiratory, cutaneous, and generalized anaphylactic symptoms). Med Clin North Am 94, 853-64. https://doi.org/10.1016/j.mcna.2010.03.005
  38. Seibert K, Zhang Y, Leahy K, Hauser S, Masferrer J, Perkins W et al. (1994) Pharmacological and biochemical demonstration of the role of cyclooxygenase 2 in inflammation and pain. Proc Natl Acad Sci USA 91, 12013-7. https://doi.org/10.1073/pnas.91.25.12013
  39. Smith WL, DeWitt DL, and Garavito RM (2000) Cyclooxygenase: structures, cellular, and molecular biology. Annu Rev Biochem 69, 145-82. https://doi.org/10.1146/annurev.biochem.69.1.145
  40. Sung YY, Kim DS, Yang WK, Nho KJ, Seo HS, Kim YS et al. (2012) Inhibitory effects of Drynaria fortunei extracts on house dust mite antigen-induced atopic dermatitis in NC/Nga mice. J Ethnopharmacol 144, 94-100. https://doi.org/10.1016/j.jep.2012.08.035
  41. Tezuka Y, Irikawa S, Kaneko T, Banskota A, Nagaoka T, Xiong Q et al. (2001) Screening of Chinese herbal drug extracts for inhibitory activity on nitric oxide production and identification of an active compound of Zanthoxylum bungeanum. J Ethnopharmacol 77, 209-17. https://doi.org/10.1016/S0378-8741(01)00300-2
  42. Towbin HT, Staehelin T, and Gordon J (1979) Electrophoretic transfers of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76, 4350-4. https://doi.org/10.1073/pnas.76.9.4350
  43. Tracey KJ, Wei H, Manogue KR, Fong Y, Hesse DG, and Nguyen HT (1988) Cachectin/tumor necrosis factor induces cachexia, anemia, and inflammation. J Exp Med 167, 1211-27. https://doi.org/10.1084/jem.167.3.1211
  44. Willeaume V, Kruys V, Mijatovic T, and Huez G (1996) Tumor necrosis factor-alpha production induced by viruses and by lipopolysaccharide in macrophages: similarities and differences. J Inflamm 46, 1-12.
  45. Yang EJ, Ham YM, Lee WJ, Lee NH, and Hyun CG (2013) Antiinflammatory effects of apo-9-fucoxanthinone from the brown alga, Sargassum muticum. J Pharm Sci 21, 1-7.
  46. Yun YG, Kim GM, Lee SJ, Ryu SH, and Jang SI (2007) Inhibitory effect of aqueous extract from Lonicera japonica flower on LPS-induced inflammatory mediators in RAW 264.7 macrophages. Kor J Herbology 22, 117-25.

Cited by

  1. Anti-Inflammatory Effect of Ethanol Extract from Grateloupia elliptica Holmes on Lipopolysaccharide-Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears vol.44, pp.8, 2015, https://doi.org/10.3746/jkfn.2015.44.8.1128
  2. Anti-Inflammatory Effect of Alginate Oligosaccharides Produced by an Alginate-Degrading Enzyme from Shewanella oneidensis PKA1008 on LPS-Induced RAW 264.7 Cells vol.48, pp.6, 2015, https://doi.org/10.5657/KFAS.2015.0888
  3. Anti-Inflammatory Effect of Grateloupia imbricata Holmes Ethanol Extract on LPS-Induced RAW 264.7 Cells vol.45, pp.2, 2016, https://doi.org/10.3746/jkfn.2016.45.2.181
  4. 잔가시 모자반 추출물의 주름 개선 및 미백 효과 vol.44, pp.1, 2016, https://doi.org/10.4014/mbl.1510.10002
  5. Sargassum Seaweed as a Source of Anti-Inflammatory Substances and the Potential Insight of the Tropical Species: A Review vol.17, pp.10, 2014, https://doi.org/10.3390/md17100590
  6. Anti-Inflammatory Effects and Their Correlation with Microbial Community of Shindari, a Traditional Jeju Beverage vol.6, pp.3, 2014, https://doi.org/10.3390/fermentation6030087
  7. Anti-Inflammatory and Antioxidant Properties of Ethanol Extracts of Raw, Blanched, Steamed, and Sous-Vide Cooked Okra (Abelmoschus esculentus L.) in LPS or H2O2-Treated RAW264.7 Cells vol.11, pp.5, 2014, https://doi.org/10.3390/app11052432
  8. Inhibition of nitric oxide and lipid accumulation by Sargassum sp. seaweeds and their antioxidant properties vol.28, pp.2, 2021, https://doi.org/10.11002/kjfp.2021.28.2.288
  9. Anti-inflammatory Effect of Hot Water Extracts of Acer tegmentosum Maxim Extracted Under Optimal Conditions for Antioxidant Activity on LPS-Induced RAW 264.7 Cells vol.32, pp.2, 2021, https://doi.org/10.7856/kjcls.2021.32.2.269
  10. Antioxidant and Anti-Inflammatory Effects of Merin Beet Sprout Extract vol.50, pp.9, 2014, https://doi.org/10.3746/jkfn.2021.50.9.921