Nutrition Research and Practice

The Korean Nutrition Society (한국영양학회)
권호 표지
DOI QR코드

DOI QR Code

Antioxidant activity and anti-inflammatory activity of ethanol extract and fractions of Doenjang in LPS-stimulated RAW 264.7 macrophages

  • Received : 2015.03.16
  • Accepted : 2015.05.14
  • Published : 2015.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

BACKGROUND/OBJECTIVES: Fermentation can increase functional compounds in fermented soybean products, thereby improving antioxidant and/or anti-inflammatory activities. We investigated the changes in the contents of phenolics and isoflavones, antioxidant activity and anti-inflammatory activity of Doenjang during fermentation and aging. MATERIALS/METHODS: Doenjang was made by inoculating Aspergillus oryzae and Bacillus licheniformis in soybeans, fermenting and aging for 1, 3, 6, 8, and 12 months (D1, D3, D6, D8, and D12). Doenjang was extracted using ethanol, and sequentially fractioned by hexane, dichloromethane (DM), ethylacetate (EA), n-butanol, and water. The contents of total phenolics, flavonoids and isoflavones, 2,2-diphenyl-1 picryl hydrazyl (DPPH) radical scavenging activity, and ferric reducing antioxidant power (FRAP) were measured. Anti-inflammatory effects in terms of nitric oxide (NO), prostaglandin (PG) E2 and pro-inflammatory cytokine production and inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 expressions were also measured using LPS-treated RAW 264.7 macrophages. RESULTS: Total phenolic and flavonoid contents showed a gradual increase during fermentation and 6 months of aging and were sustained thereafter. DPPH radical scavenging activity and FRAP were increased by fermentation. FRAP was further increased by aging, but DPPH radical scavenging activity was not. Total isoflavone and glycoside contents decreased during fermentation and the aging process, while aglycone content and its proportion increased up to 3 or 6 months of aging and then showed a slow decrease. DM and EA fractions of Doenjang showed much higher total phenolic and flavonoid contents, and DPPH radical scavenging activity than the others. At $100{\mu}g/mL$, DM and EA fractions of D12 showed strongly suppressed NO production to 55.6% and 52.5% of control, respectively, and PGE2 production to 25.0% and 28.3% of control with inhibition of iNOS or COX-2 protein expression in macrophages. CONCLUSIONS: Twelve month-aged Doenjang has potent antioxidant and anti-inflammatory activities with high levels of phenolics and isoflavone aglycones, and can be used as a beneficial food for human health.

Keywords

References

  1. Lee TH, Do MH, Oh YL, Cho DW, Kim SH, Kim SY. Dietary fermented soybean suppresses UVB-induced skin inflammation in hairless mice via regulation of the MAPK signaling pathway. J Agric Food Chem 2014;62:8962-72. https://doi.org/10.1021/jf5018252
  2. Kwon DY, Daily JW 3rd, Kim HJ, Park S. Antidiabetic effects of fermented soybean products on type 2 diabetes. Nutr Res 2010;30:1-13. https://doi.org/10.1016/j.nutres.2009.11.004
  3. Chai C, Ju HK, Kim SC, Park JH, Lim J, Kwon SW, Lee J. Determination of bioactive compounds in fermented soybean products using GC/MS and further investigation of correlation of their bioactivities. J Chromatogr B Analyt Technol Biomed Life Sci 2012;880:42-9. https://doi.org/10.1016/j.jchromb.2011.11.013
  4. Chung SI, Rico CW, Kang MY. Comparative study on the hypoglycemic and antioxidative effects of fermented paste (doenjang) prepared from soybean and brown rice mixed with rice bran or red ginseng marc in mice fed with high fat diet. Nutrients 2014;6:4610-24. https://doi.org/10.3390/nu6104610
  5. Hu CC, Hsiao CH, Huang SY, Fu SH, Lai CC, Hong TM, Chen HH, Lu FJ. Antioxidant activity of fermented soybean extract. J Agric Food Chem 2004;52:5735-9. https://doi.org/10.1021/jf035075b
  6. Chen YC, Sugiyama Y, Abe N, Kuruto-Niwa R, Nozawa R, Hirota A. DPPH radical-scavenging compounds from Dou-chi, a soybean fermented food. Biosci Biotechnol Biochem 2005;69:999-1006. https://doi.org/10.1271/bbb.69.999
  7. Kwak CS, Lee MS, Park SC. Higher antioxidant properties of Chungkookjang, a fermented soybean paste, may be due to increased aglycone and malonylglycoside isoflavone during fermentation. Nutr Res 2007;27:719-27. https://doi.org/10.1016/j.nutres.2007.09.004
  8. Jung KO, Park SY, Park KY. Longer aging time increases the anticancer and antimetastatic properties of doenjang. Nutrition 2006;22:539-45. https://doi.org/10.1016/j.nut.2005.11.007
  9. Park KY, Jung KO, Rhee SH, Choi YH. Antimutagenic effects of doenjang (Korean fermented soypaste) and its active compounds. Mutat Res 2003;523-524:43-53. https://doi.org/10.1016/S0027-5107(02)00320-2
  10. Park KJ, Kim YM, Lee BH, Lee BK. Fungal microflora on Korean home-made Meju. Korean J Mycol 1977;5:7-12.
  11. Lee SS. Meju fermentation for a raw material of Korean traditional soy products. Korean J Mycol 1995;23:161-75.
  12. Kwak CS, Park SC, Song KY. Doenjang, a fermented soybean paste, decreased visceral fat accumulation and adipocyte size in rats fed with high fat diet more effectively than nonfermented soybeans. J Med Food 2012;15:1-9. https://doi.org/10.1089/jmf.2010.1224
  13. Park NY, Rico CW, Lee SC, Kang MY. Comparative effects of doenjang prepared from soybean and brown rice on the body weight and lipid metabolism in high fat-fed mice. J Clin Biochem Nutr 2012;51:235-40.
  14. Cha YS, Yang JA, Back HI, Kim SR, Kim MG, Jung SJ, Song WO, Chae SW. Visceral fat and body weight are reduced in overweight adults by the supplementation of Doenjang, a fermented soybean paste. Nutr Res Pract 2012;6:520-6. https://doi.org/10.4162/nrp.2012.6.6.520
  15. Blay M, Espinel AE, Delgado MA, Baiges I, Blade C, Arola L, Salvado J. Isoflavone effect on gene expression profile and biomarkers of inflammation. J Pharm Biomed Anal 2010;51:382-90. https://doi.org/10.1016/j.jpba.2009.03.028
  16. Scott A, Khan KM, Cook JL, Duronio V. What is "inflammation"? Are we ready to move beyond Celsus? Br J Sports Med 2004;38:248-9. https://doi.org/10.1136/bjsm.2003.011221
  17. Prasad S, Phromnoi K, Yadav VR, Chaturvedi MM, Aggarwal BB. Targeting inflammatory pathways by flavonoids for prevention and treatment of cancer. Planta Med 2010;76:1044-63. https://doi.org/10.1055/s-0030-1250111
  18. Witkamp R, Monshouwer M. Signal transduction in inflammatory processes, current and future therapeutic targets: a mini review. Vet Q 2000;22:11-6. https://doi.org/10.1080/01652176.2000.9695016
  19. Bak MJ, Jeong JH, Kang HS, Jin KS, Ok S, Jeong WS. Cedrela sinensis leaves suppress oxidative stress and expressions of iNOS and COX-2 via MAPK signaling pathways in RAW 264.7 cells. J Food Sci Nutr 2009;14:269-76. https://doi.org/10.3746/jfn.2009.14.4.269
  20. Kwon SH, Lee KB, Im KS, Kim SO, Park KY. Weight reduction and lipid lowering effects of Korean traditional soybean fermented products. J Korean Soc Food Sci Nutr 2006;35:1194-9. https://doi.org/10.3746/jkfn.2006.35.9.1194
  21. Jang CH, Lim JK, Kim JH, Park CS, Kwon DY, Kim YS, Shin DH, Kim JS. Change of isoflavone content during manufacturing of Cheonggukjang, a traditional Korean fermented soyfood. Food Sci Biotechnol 2006;15:643-6.
  22. Nakajima N, Nozaki N, Ishihara K, Ishikawa A, Tsuji H. Analysis of isoflavone content in tempeh, a fermented soybean, and preparation of a new isoflavone-enriched tempeh. J Biosci Bioeng 2005;100:685-7. https://doi.org/10.1263/jbb.100.685
  23. Shon MY, Seo KI, Lee SW, Choi SH, Sung NJ. Biological activities of Chungkugjang prepared with black bean and changes in phytoestrogen content during fermentation. Korean J Food Sci Technol 2000;32:936-41.
  24. Danciu C, Soica C, Csanyi E, Ambrus R, Feflea S, Peev C, Dehelean C. Changes in the anti-inflammatory activity of soy isoflavonoid genistein versus genistein incorporated in two types of cyclodextrin derivatives. Chem Cent J 2012;6:58. https://doi.org/10.1186/1752-153X-6-58
  25. Liang YC, Huang YT, Tsai SH, Lin-Shiau SY, Chen CF, Lin JK. Suppression of inducible cyclooxygenase and inducible nitric oxide synthase by apigenin and related flavonoids in mouse macrophages. Carcinogenesis 1999;20:1945-52. https://doi.org/10.1093/carcin/20.10.1945
  26. Ji G, Zhang Y, Yang Q, Cheng S, Hao J, Zhao X, Jiang Z. Genistein suppresses LPS-induced inflammatory response through inhibiting $NF-{\kappa}B$ following AMP kinase activation in RAW 264.7 macrophages. PLoS One 2012;7:e53101. https://doi.org/10.1371/journal.pone.0053101
  27. Singleton VL, Orthofer R, Lamuela-Raventos RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods Enzymol 1999;299:152-78. https://doi.org/10.1016/S0076-6879(99)99017-1
  28. Chae SK, Kang GS, Ma SJ, Bang KW, Oh MW, Oh SH. Standard Food Analysis. Seoul: Jigu Publishing Co.; 1997.
  29. Senba Y, Nishishita T, Saito K, Yoshioka H, Yoshioka H. Stopped-flow and spectrophotometric study on radical scavenging by tea catechins and the model compounds. Chem Pharm Bull (Tokyo) 1999;47:1369-74. https://doi.org/10.1248/cpb.47.1369
  30. Yildirim A, Mavi A, Kara AA. Determination of antioxidant and antimicrobial activities of Rumex crispus L. extracts. J Agric Food Chem 2001;49:4083-9. https://doi.org/10.1021/jf0103572
  31. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63. https://doi.org/10.1016/0022-1759(83)90303-4
  32. Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem 1982;126:131-8. https://doi.org/10.1016/0003-2697(82)90118-X
  33. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-54. https://doi.org/10.1016/0003-2697(76)90527-3
  34. Zhang R, Brennan ML, Shen Z, MacPherson JC, Schmitt D, Molenda CE, Hazen SL. Myeloperoxidase functions as a major enzymatic catalyst for initiation of lipid peroxidation at sites of inflammation. J Biol Chem 2002;277:46116-22. https://doi.org/10.1074/jbc.M209124200
  35. Coulibaly AY, Kiendrebeogo M, Kehoe PG, Sombie PA, Lamien CE, Millogo JF, Nacoulma OG. Antioxidant and anti-inflammatory effects of Scoparia dulcis L. J Med Food 2011;14:1576-82. https://doi.org/10.1089/jmf.2010.0191
  36. Chang ST, Wu JH, Wang SY, Kang PL, Yang NS, Shyur LF. Antioxidant activity of extracts from Acacia confusa bark and heartwood. J Agric Food Chem 2001;49:3420-4. https://doi.org/10.1021/jf0100907
  37. Michel P, Dobrowolska A, Kicel A, Owczarek A, Bazylko A, Granica S, Piwowarski JP, Olszewska MA. Polyphenolic profile, antioxidant and anti-inflammatory activity of eastern teaberry (Gaultheria procumbens L.) leaf extracts. Molecules 2014;19:20498-520. https://doi.org/10.3390/molecules191220498
  38. Noh KH, Jang JH, Min KH, Chinzorig R, Lee MO, Song YS. Suppressive effect of green tea seed coat ethyl acetate fraction on inflammation and its mechanism in RAW264.7 macrophage cell. J Korean Soc Food Sci Nutr 2011;40:625-34. https://doi.org/10.3746/jkfn.2011.40.5.625
  39. Kim YS, Lee SJ, Hwang JW, Kim EH, Park PJ, Jeong JH. Anti-inflammatory effects of extracts from Ligustrum ovalifolium H. leaves on RAW264.7 macrophages. J Korean Soc Food Sci Nutr 2012;41:1205-10. https://doi.org/10.3746/jkfn.2012.41.9.1205
  40. Velioglu YS, Mazza G, Gao L, Oomah BD. Antioxidant activity and total phenolics in selected fruits, vegetables, and grain products. J Agric Food Chem 1998;46:4113-7. https://doi.org/10.1021/jf9801973
  41. Zainol MK, Abd-Hamid A, Yusof S, Muse R. Antioxidative activity and total phenolic compounds of leaf, root and petiole of four accessions of Centella asiatica (L.) urban. Food Chem 2003;81:575-81. https://doi.org/10.1016/S0308-8146(02)00498-3
  42. Kwak CS, Kim SA, Lee MS. The correlation of antioxidative effects of 5 Korean common edible seaweeds and total polyphenol content. J Korean Soc Food Sci Nutr 2005;34:1143-50. https://doi.org/10.3746/jkfn.2005.34.8.1143
  43. Setchell KD, Brown NM, Zimmer-Nechemias L, Brashear WT, Wolfe BE, Kirschner AS, Heubi JE. Evidence for lack of absorption of soy isoflavone glycosides in humans, supporting the crucial role of intestinal metabolism for bioavailability. Am J Clin Nutr 2002;76:447-53. https://doi.org/10.1093/ajcn/76.2.447
  44. Izumi T, Piskula MK, Osawa S, Obata A, Tobe K, Saito M, Kataoka S, Kubota Y, Kikuchi M. Soy isoflavone aglycones are absorbed faster and in higher amounts than their glucosides in humans. J Nutr 2000;130:1695-9. https://doi.org/10.1093/jn/130.7.1695
  45. Pratt DE. Natural antioxidants of soybeans and other oil-seeds. In: Simic MG, Karel M, editors. Autoxidation in Food and Biological Systems. New York (NY): Plenum Press; 1980. p.283-293.
  46. Sheih IC, Fang TJ, Wu TK, Chen RY. Effects of fermentation on antioxidant properties and phytochemical composition of soy germ. J Sci Food Agric 2014;94:3163-70. https://doi.org/10.1002/jsfa.6666
  47. Dziedzic SZ, Hudson BJ. Hydroxy isoflavones as antioxidants for edible oils. Food Chem 1983;11:161-6. https://doi.org/10.1016/0308-8146(83)90099-7
  48. Kwon SH, Shon MY. Antioxidant and anticarcinogenic effects of traditional Doenjang during maturation periods. Korean J Food Preserv 2004;11:461-7.
  49. Kim MH, Im SS, Kim SH, Kim GE, Lee JH. Antioxidative materials in domestic Meju and Doenjang 2. Seperation of lipophilic brown pigment and their antioxidative activity. J Korean Soc Food Sci Nutr 1994;23:251-60.
  50. Jeong JK, Zheng Y, Choi HS, Han GJ, Park KY. Catabolic enzyme activities and physiological functionalities of lactic acid bacteria isolated from Korean traditional Meju. J Korean Soc Food Sci Nutr 2010;39:1854-9. https://doi.org/10.3746/jkfn.2010.39.12.1854
  51. Roh C, Jung U, Jo SK. 6,7,4'-Trihydroxyisoflavone from Doenjang inhibits lipid accumulation. Food Chem 2011;129:183-7. https://doi.org/10.1016/j.foodchem.2011.04.082
  52. Park JS, Park HY, Kim DH, Kim DH, Kim HK. Ortho-dihydroxyisoflavone derivatives from aged Doenjang (Korean fermented soypaste) and its radical scavenging activity. Bioorg Med Chem Lett 2008;18:5006-9. https://doi.org/10.1016/j.bmcl.2008.08.016
  53. Wang HJ, Murphy PA. Mass balance study of isoflavones during soybean processing. J Agric Food Chem 1996;44:2377-83. https://doi.org/10.1021/jf950535p
  54. Wu CH, Chou CC. Enhancement of aglycone, vitamin K2 and superoxide dismutase activity of black soybean through fermentation with Bacillus subtilis BCRC 14715 at different temperatures. J Agric Food Chem 2009;57:10695-700. https://doi.org/10.1021/jf902752t
  55. Lee S, Seo MH, Oh DK, Lee CH. Targeted metabolomics for Aspergillus oryzae-mediated biotransformation of soybean isoflavones, showing variations in primary metabolites. Biosci Biotechnol Biochem 2014;78:167-74. https://doi.org/10.1080/09168451.2014.877827
  56. Jeong JW, Lee HH, Han MH, Kim GY, Kim WJ, Choi YH. Anti-inflammatory effects of genistein via suppression of the toll-like receptor 4-mediated signaling pathway in lipopolysaccharide-stimulated BV2 microglia. Chem Biol Interact 2014;212:30-9. https://doi.org/10.1016/j.cbi.2014.01.012
  57. Sung YH, Chang HK, Kim SE, Kim YM, Seo JH, Shin MC, Shin MS, Yi JW, Shin DH, Kim H, Kim CJ. Anti-inflammatory and analgesic effects of the aqueous extract of Corni fructus in murine RAW 264.7 macrophage cells. J Med Food 2009;12:788-95. https://doi.org/10.1089/jmf.2008.1011
  58. Bredt DS, Snyder SH. Nitric oxide: a physiologic messenger molecule. Annu Rev Biochem 1994;63:175-95. https://doi.org/10.1146/annurev.bi.63.070194.001135
  59. 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 2003;139(1):11-20. https://doi.org/10.1038/sj.bjp.0705231
  60. Lee EJ, Kim C, Kim JY, Kim SM, Nam D, Jang HJ, Kim SH, Shim BS, Ahn KS, Choi SH, Jung SH, Ahn KS. Inhibition of LPS-induced inflammatory biomarkers by ethyl acetate fraction of Patrinia scabiosaefolia through suppression of $NF-{\kappa}B$ activation in RAW 264.7 cells. Immunopharmacol Immunotoxicol 2012;34:282-91. https://doi.org/10.3109/08923973.2011.602412
  61. Koeberle A, Northoff H, Werz O. Curcumin blocks prostaglandin E2 biosynthesis through direct inhibition of the microsomal prostaglandin E2 synthase-1. Mol Cancer Ther 2009;8:2348-55.

Cited by

  1. ) Ingestion in Estrogen-Deficient Rats vol.83, pp.8, 2018, https://doi.org/10.1111/1750-3841.14214
  2. 된장과 고추장 키위장아찌의 기능성 및 관능 특성 vol.50, pp.2, 2015, https://doi.org/10.9721/kjfst.2018.50.2.238
  3. Doenjang, A Korean Traditional Fermented Soybean Paste, Ameliorates Neuroinflammation and Neurodegeneration in Mice Fed a High-Fat Diet vol.11, pp.8, 2019, https://doi.org/10.3390/nu11081702
  4. Sargassum Seaweed as a Source of Anti-Inflammatory Substances and the Potential Insight of the Tropical Species: A Review vol.17, pp.10, 2015, https://doi.org/10.3390/md17100590
  5. Discovery and Development of Inflammatory Inhibitors from 2-Phenylchromonone (Flavone) Scaffolds vol.20, pp.None, 2015, https://doi.org/10.2174/1568026620666200924115611
  6. Therapeutic effects of polyphenols in fermented soybean and black soybean products vol.81, pp.None, 2021, https://doi.org/10.1016/j.jff.2021.104467
  7. Metabolomic-Based Comparison of Traditional and Industrial Doenjang Samples with Antioxidative Activities vol.10, pp.6, 2015, https://doi.org/10.3390/foods10061377
  8. Korean traditional foods as antiviral and respiratory disease prevention and treatments: A detailed review vol.116, pp.None, 2021, https://doi.org/10.1016/j.tifs.2021.07.037
  9. Current perspectives on the anti-inflammatory potential of fermented soy foods vol.152, pp.None, 2015, https://doi.org/10.1016/j.foodres.2021.110922