Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Anti-inflammatory effects of ethyl acetate fraction of unripe astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) on lipopolysaccharide-stimulated RAW 264.7 cells

지방질다당류로 자극한 RAW 264.7 세포에서 청도반시 땡감 에틸 아세테이트 분획물의 항염증 효과

  • Park, Ye Bin (Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Jeong, Ha-Ram (Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Lee, Seung Hwan (Department of Food Science and Biotechnology, Andong National University) ;
  • Kim, Taewan (Department of Food Science and Biotechnology, Andong National University) ;
  • Kim, Dae-Ok (Department of Food Science and Biotechnology, Kyung Hee University)
  • 박예빈 (경희대학교 식품생명공학과) ;
  • 정하람 (경희대학교 식품생명공학과) ;
  • 이승환 (안동대학교 식품생명공학과) ;
  • 김태완 (안동대학교 식품생명공학과) ;
  • 김대옥 (경희대학교 식품생명공학과)
  • Received : 2018.12.28
  • Accepted : 2019.02.02
  • Published : 2019.02.28
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Abstract

Unripe astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) is a by-product produced when thinning out the superfluous fruit of persimmon. We investigated whether unripe astringent persimmon has antioxidative and anti-inflammatory effects. Unripe astringent persimmon extract was fractionated sequentially in n-hexane, chloroform, ethyl acetate, n-butanol, and water. The ethyl acetate fraction had the highest total phenolic content, total flavonoid content, and antioxidant capacity compared to those of the other fractions. Pretreatment of lipopolysaccharide-stimulated RAW 264.7 macrophages with the ethyl acetate fraction reduced nitric oxide, interleukin-6, and intracellular oxidative stress in a dose-dependent manner. Ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry analysis revealed gallic acid, protocatechuic acid, 4-hydroxybenzoic acid, quercetin-3-O-glucoside, quercetin, and p-coumaric acid as the phenolic compounds of the ethyl acetate fraction. Collectively, these findings suggest that unripe astringent persimmon is a source of functional materials that can promote antioxidative and anti-inflammatory effects.

본 연구에서는 적과한 청도반시 땡감을 40% (v/v) 에탄올-물 혼합 용액으로 추출하고, 이 추출물을 액체-액체 추출법으로 5개 분획물(노말-헥세인, 클로로폼, 에틸 아세테이트, 노말-뷰탄올, 물)을 얻었다. 5개 분획물 중에서 에틸 아세테이트 분획물이 총 페놀 함량, 총 플라보노이드 함량 및 산화방지능이 가장 높았다. 에틸 아세테이트 분획물은 지방질다당류로 자극한 RAW 264.7 세포에서 산화 질소, IL-6, 산화 스트레스를 감소시켰다. UPLC-ESI-MS/MS를 이용하여 땡감 에틸 아세테이트 분획물에서 페놀 화합물로 갈산, 프로토카테츄산, 4-하이드록시벤조산, 쿼서틴-3-O-글루코사이드, 파라-쿠마르산, 쿼서틴을 동정하였다. 본 연구를 통해서 적과 과정에서 부산물로 버려지는 청도반시 땡감이 산화방지능 및 항염증 효과를 보유한 기능성 소재로서 활용될 가능성을 제시하였다. 그러나 향후 적과한 청도반시 땡감의 산화방지능에 기반한 항염증 효과를 좀 더 명확하게 밝히기 위해서는 청도반시 땡감에 존재하는 개별 생리활성물질의 정량 분석 및 항염증에 대한 분자생물학적 메커니즘을 규명할 필요가 있다.

Keywords

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Fig. 1. Inhibitory effects of the ethyl acetate fraction from unripe astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo- Bansi) on nitrite production in lipopolysaccharide (LPS)- stimulated RAW 264.7 cells.

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Fig. 2. Inhibitory effects of the ethyl acetate fraction from unripe astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo- Bansi) on inflammatory cytokine interleukin 6 (IL-6) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells.

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Fig. 3. Effects of the ethyl acetate fraction from unripe astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo- Bansi) on intracellular oxidative stress in RAW 264.7 cells against oxidative stress induced with AAPH using the DCFHDA assay.

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Fig. 4. UPLC-ESI-MS/MS chromatogram of the ethyl acetate fraction from unripe astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi).

Table 1. Total phenolic and flavonoid contents, and antioxidant capacities of five fractions of unripe astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi)

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Table 2. Identification of phenolic compounds in the ethyl acetate fraction of unripe astringent persimmon (Diospyros kaki Thunb. cv.Cheongdo-Bansi) using UPLC-ESI-MS/MS

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References

  1. Azad N, Rojanasakul Y, Vallyathan V. Inflammation and lung cancer: roles of reactive oxygen/nitrogen species. J. Toxicol. Environ. Health Part B 11: 1-15 (2008) https://doi.org/10.1080/10937400701436460
  2. Bennett RN, Wallsgrove RM. Secondary metabolites in plant defence mechanisms. New Phytol. 127: 617-633 (1994) https://doi.org/10.1111/j.1469-8137.1994.tb02968.x
  3. Biswas SK. Does the interdependence between oxidative stress and inflammation explain the antioxidant paradox? Oxid. Med. Cell. Longev. 2016: 5698931 (2016) https://doi.org/10.1155/2016/5698931
  4. Brand-Williams W, Cuvelier ME, Berset C. Use of a free radical method to evaluate antioxidant activity. LWT-Food Sci. Technol. 28: 25-30 (1995) https://doi.org/10.1016/S0023-6438(95)80008-5
  5. Butt MS, Sultan MT, Aziz M, Naz A, Ahmed W, Kumar N, Imran M. Persimmon (Diospyros kaki) fruit: hidden phytochemicals and health claims. EXCLI J. 14: 542-561 (2015)
  6. Chen XN, Fan JF, Yue X, Wu XR, Li LT. Radical scavenging activity and phenolic compounds in persimmon (Diospyros kaki L. cv. Mopan). J. Food Sci. 73: C24-C28 (2008) https://doi.org/10.1111/j.1750-3841.2007.00587.x
  7. Coleman JW. Nitric oxide in immunity and inflammation. Int. Immunopharmacol. 1: 1397-1406 (2001) https://doi.org/10.1016/S1567-5769(01)00086-8
  8. Comalada M, Ballester I, Bailon E, Sierra S, Xaus J, Galvez J, Sanchez de Medina F, Zarzuelo A. Inhibition of pro-inflammatory markers in primary bone marrow-derived mouse macrophages by naturally occurring flavonoids: analysis of the structure-activity relationship. Biochem. Pharmacol. 72: 1010-1021 (2006) https://doi.org/10.1016/j.bcp.2006.07.016
  9. Comalada M, Camuesco D, Sierra S, Ballester I, Xaus J, Galvez J, Zarzuelo A. In vivo quercitrin anti-inflammatory effect involves release of quercetin, which inhibits inflammation through downregulation of the NF-${\kappa}$B pathway. Eur. J. Immunol. 35: 584-592 (2005) https://doi.org/10.1002/eji.200425778
  10. Heo H-J, Cho H-Y, Hong B, Kim H-K, Kim E-K, Kim B-G, Shin DH. Protective effect of 4',5-dihydroxy-3',6,7-trimethoxyflavone from Artemisia asiatica against A${\beta}$-induced oxidative stress in PC12 cells. Amyloid-J. Protein Fold. Disord. 8: 194-201 (2001) https://doi.org/10.3109/13506120109007362
  11. Hossain A, Moon HK, Kim J-K. Antioxidant properties of Korean major persimmon (Diospyros kaki) leaves. Food Sci. Biotechnol. 27: 177-184 (2018) https://doi.org/10.1007/s10068-017-0195-y
  12. Hussain SP, Hofseth LJ, Harris CC. Radical causes of cancer. Nat. Rev. Cancer 3: 276-285 (2003) https://doi.org/10.1038/nrc1046
  13. Hussain T, Tan B, Yin Y, Blachier F, Tossou MCB, Rahu N. Oxidative stress and inflammation: what polyphenols can do for us? Oxid. Med. Cell. Longev. 2016: 7432797 (2016)
  14. Janssens S, Beyaert R. Role of Toll-like receptors in pathogen recognition. Clin. Microbiol. Rev. 16: 637-646 (2003) https://doi.org/10.1128/CMR.16.4.637-646.2003
  15. Jeon IH, Kang HJ, Lee H-S, Shin JH, Park YG, Jeong S-I, Jang SI. Antioxidant and anti-inflammatory activities of water-soluble extracts from different parts of Kojongsi persimmon (Diospyros kaki L.). Korean J. Food Sci. Technol. 46: 505-510 (2014) https://doi.org/10.9721/KJFST.2014.46.4.505
  16. Jeong D-W, Cho CH, Lee JS, Lee SH, Kim T, Kim D-O. Deastringent peel extracts of persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) protect neuronal PC-12 and SH-SY5Y cells against oxidative stress. J. Microbiol. Biotechnol. 28: 1094-1104 (2018) https://doi.org/10.4014/jmb.1801.01013
  17. Kim D-O, Jeong SW, Lee CY. Antioxidant capacity of phenolic phytochemicals from various cultivars of plums. Food Chem. 81: 321-326 (2003) https://doi.org/10.1016/S0308-8146(02)00423-5
  18. Kim D-O, Lee KW, Lee HJ, Lee CY. Vitamin C equivalent antioxidant capacity (VCEAC) of phenolic phytochemicals. J. Agric. Food Chem. 50: 3713-3717 (2002) https://doi.org/10.1021/jf020071c
  19. Kim EO, Lee H, Cho CH, Kim YJ, Kim D-O. Antioxidant capacity and anti-inflammatory effect of the ethyl acetate fraction of dried persimmon (Diospyros kaki Thumb.) on THP-1 human acute monocytic leukemia cell line. J. Korean Soc. Appl. Biol. Chem. 54: 606-611 (2011a)
  20. Kim H-J, Park T-S, Jung M-S, Son J-H. Study on the anti-oxidant and anti-inflammatory activities of sarcocarp and calyx of persimmon (Cheongdo Bansi). J. Appl. Biol. Chem. 54: 71-78 (2011b) https://doi.org/10.3839/jabc.2011.013
  21. Lee MS, Lee II, Kim Y, Kim YJ, Heo HJ, Kim D-O. Inhibitory effect of the ethyl acetate fraction from astringent persimmon on $H_2O_2$-induced oxidative stress in HepG2 cells. Food Sci. Biotechnol. 23: 1247-1252 (2014) https://doi.org/10.1007/s10068-014-0171-8
  22. Mallavadhani UV, Panda AK, Rao YR. Pharmacology and chemotaxonomy of diospyros. Phytochemistry 49: 901-951 (1998) https://doi.org/10.1016/S0031-9422(97)01020-0
  23. Park W. Inhibitory effect of gallic acid on production of interleukins in mouse macrophage stimulated by lipopolysaccharide. J. Pharmacopuncture 13: 63-71 (2010)
  24. Ryter SW, Kim HP, Hoetzel A, Park JW, Nakahira K, Wang X, Choi AMK. Mechanisms of cell death in oxidative stress. Antioxid. Redox Signal. 9: 49-89 (2007) https://doi.org/10.1089/ars.2007.9.49
  25. Schieber A, Stintzing FC, Carle R. By-products of plant food processing as a source of functional compounds-recent developments. Trends Food Sci. Technol. 12: 401-413 (2001) https://doi.org/10.1016/S0924-2244(02)00012-2
  26. Shin D-J, Kim K-H, Son G-M, Lee S-C, Hwang Y-I. Changes of physicochemical properties during preparation of prepersimmon pickles. J. Korean Soc. Food Sci. Nutr. 29: 420-424 (2000)
  27. Singleton VL, Rossi JA, Jr. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am. J. Enol. Vitic. 16: 144-158 (1965)
  28. Suzuki T, Someya S, Hu F, Tanokura M. Comparative study of catechin compositions in five Japanese persimmons (Diospyros kaki). Food Chem. 93: 149-152 (2005) https://doi.org/10.1016/j.foodchem.2004.10.017
  29. Torres JL, Varela B, Garcia MT, Carilla J, Matito C, Centelles JJ, Cascante M, Sort X, Bobet R. Valorization of grape (Vitis vinifera) byproducts. Antioxidant and biological properties of polyphenolic fractions differing in procyanidin composition and flavonol content. J. Agric. Food Chem. 50: 7548-7555 (2002) https://doi.org/10.1021/jf025868i
  30. Winter AN, Brenner MC, Punessen N, Snodgrass M, Byars C, Arora Y, Linseman DA. Comparison of the neuroprotective and antiinflammatory effects of the anthocyanin metabolites, protocatechuic acid and 4-hydroxybenzoic acid. Oxid. Med. Cell. Longev. 2017: 6297080 (2017)