Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
권호 표지
DOI QR코드

DOI QR Code

Antioxidant capacity and hepatoprotective effect of ethyl acetate fraction from shoot of Aralia elata on alcohol-induced cytotoxicity

두릅 아세트산 에틸 분획물의 산화방지 효과 및 알코올에 대한 간세포 보호효과

  • Kwon, Bong Seok (Division of Applied Life Science (BK21 plus), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Park, Seon Kyeong (Division of Applied Life Science (BK21 plus), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Kim, Jong Min (Division of Applied Life Science (BK21 plus), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Kang, Jin Yong (Division of Applied Life Science (BK21 plus), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Park, Sang Hyun (Division of Applied Life Science (BK21 plus), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Kang, Jeong Eun (Division of Applied Life Science (BK21 plus), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Lee, Chang Jun (Division of Applied Life Science (BK21 plus), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Park, Su Bin (Division of Applied Life Science (BK21 plus), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Yoo, Seul Ki (Division of Applied Life Science (BK21 plus), Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Lee, Uk (Division of Special Forest Products, National Institute of Forest Science) ;
  • Heo, Ho Jin (Division of Applied Life Science (BK21 plus), Institute of Agriculture and Life Science, Gyeongsang National University)
  • 권봉석 (경상대학교 응용생명과학부(BK21 plus), 농업생명과학연구원) ;
  • 박선경 (경상대학교 응용생명과학부(BK21 plus), 농업생명과학연구원) ;
  • 김종민 (경상대학교 응용생명과학부(BK21 plus), 농업생명과학연구원) ;
  • 강진용 (경상대학교 응용생명과학부(BK21 plus), 농업생명과학연구원) ;
  • 박상현 (경상대학교 응용생명과학부(BK21 plus), 농업생명과학연구원) ;
  • 강정은 (경상대학교 응용생명과학부(BK21 plus), 농업생명과학연구원) ;
  • 이창준 (경상대학교 응용생명과학부(BK21 plus), 농업생명과학연구원) ;
  • 박수빈 (경상대학교 응용생명과학부(BK21 plus), 농업생명과학연구원) ;
  • 유슬기 (경상대학교 응용생명과학부(BK21 plus), 농업생명과학연구원) ;
  • 이욱 (국립산림과학원 산림소득자원연구과) ;
  • 허호진 (경상대학교 응용생명과학부(BK21 plus), 농업생명과학연구원)
  • Received : 2018.01.03
  • Accepted : 2018.01.25
  • Published : 2018.04.30
Download PDF
⟨ Previous Next ⟩

Abstract

To evaluate physiological effect of Aralia elata, in vitro antioxidant activity and hepatic protective effects were investigated. Ethyl acetate fraction from Aralia elata (EFAE) had higher total phenolic content than other fractions (n-hexane, chloroform, and distilled water layers). EFAE also showed significantly greater radical scavenging activity against 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH), than other fractions. Moreover, EFAE showed dose-dependent inhibitory effect of malondialdehyde (MDA). Hepatoprotective effects of EFAE against ethanol- and $H_2O_2$-induced oxidative stress and cytotoxicity in H4IIE and HepG2 hepatic cells were examined using 2',7'-dichlorofluorescein diacetate (DCF-DA) and 3-[4,5-dimethythiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. The results showed that EFAE reduced cellular oxidative stress, and increased hepatic cell viability. In addition, EFAE inhibited ethanol-induced lipid accumulation in HepG2 cells. Finally, physiological substances of EFAE were analyzed using high performance liquid chromatography (HPLC), and the major bioactive compounds identified were 3,5-dicaffeoylquinic acid and chlorogenic acid.

본 연구에서는 두릅(shoot of Aralia elata)의 분획물이 가지는 산화방지 효과와 간세포 보호효과를 확인하고 이에 영향을 주는 생리활성물질을 분석하고자 하였다. 두릅의 각 분획물을 이용하여 총 페놀 함량, 산화방지 능력(ABTS와 DPPH 라디칼 제거 활성)의 측정을 통해 아세트산 에틸 분획물이 다른 분획물 대비 가장 높은 산화방지 능력을 가진 것을 확인하였다. 두릅의 아세트산 에틸 분획물을 이용하여 지방질과산화물의 생성 억제 능력을 확인한 결과 역시 우수한 억제 활성효과를 나타내었다. 그리고 두릅 아세트산 에틸 분획물은 간세포(H4IIE와 HepG2)에서 에탄올과 과산화수소에 의한 강한 산화적 스트레스를 억제하는 효과를 나타내었으며, 생존율 또한 증가시키는 것으로 나타났다. 또한 HepG2 세포에서 알코올에 의한 지방질 축적을 유도하였을 때 두릅의 아세트산 에틸 분획물이 지방질 축적의 억제 효과도 가지는 것으로 나타났다. 이러한 두릅 아세트산 에틸 분획물의 생리활성물질을 확인하기 위해 HPLC를 분석한 결과 클로로겐산과 3,5-다이카페오일퀸산이 주요 생리활성물질임을 확인하였다. 본 연구의 결과를 바탕으로 고려할 때, 두릅의 아세트산 에틸 분획물은 우수한 산화방지 능력과 간세포 보호 능력을 나타냄에 따라 간 질환을 예방할 수 있는 천연물 유래 건강기능식품 소재로서 활용될 수 있을 것으로 기대된다.

Keywords

References

  1. Bak MJ, Jun M, Jeong WS. Antioxidant and hepatoprotective effects of the red ginseng essential oil in $H_2O_2$-treated HepG2 cells and $CCl_4$-treated mice. Int. J. Mol. Sci. 13: 2314-2330 (2012) https://doi.org/10.3390/ijms13022314
  2. Boucher PD, Hines RN. In vitro binding and functional studies comparing the human CYPIAI negative regulatory element with the orthologous sequences from rodent genes. Carcinogenesis. 16: 383-392 (1995) https://doi.org/10.1093/carcin/16.2.383
  3. Brandon-Warner E, Sugg JA, Schrum LW, McKillop IH. Silibinin inhibits ethanol metabolism and ethanol-dependent cell proliferation in an in vitro model of hepatocellular carcinoma. Cancer lett. 291: 120-129 (2010) https://doi.org/10.1016/j.canlet.2009.10.004
  4. Cha JY, Ahn HY, Eom KE, Park BK, Jun BS, Cho YS. Antioxidative activity of Aralia elata shoot and leaf extracts. J. Life Sci. 19: 652-658 (2009) https://doi.org/10.5352/JLS.2009.19.5.652
  5. 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. 49: 3420-3424 (2001) https://doi.org/10.1021/jf0100907
  6. Cho AS, Jeon SM, Kim MJ, Yeo J, Seo KI, Choi MS, Lee MK. Chlorogenic acid exhibits anti-obesity property and improves lipid metabolism in high-fat diet-induced-obese mice. Food Chem. Toxicol. 48: 937-943 (2010) https://doi.org/10.1016/j.fct.2010.01.003
  7. Chung CK, Jung ME. Ethanol fraction of Aralia elata Seemann enhances antioxidant activity and lowers serum lipids in rats when administered with benzo(a)pyrene. Bio. Pharm. Bull. 26: 1502-1504 (2003) https://doi.org/10.1248/bpb.26.1502
  8. Dudonne S, Vitrac X, Coutiere P, Woillez M, Merillon J. Comparative study of antioxidant properties and total phenolic content of 30 plant extracts of industrial interest using DPPH, ABTS, FRAP, SOD, and ORAC assays. J. Agric. Food Chem. 57: 1768-1774 (2009) https://doi.org/10.1021/jf803011r
  9. Ela MAA, El-Lakany AM, Abdel-Kader MS, Alqasoumi SI, Shams- El-Din SM, Hammoda HM. New quinic acid derivatives from hepatoprotective Inula Crithmoides root extract. Helv. Chim. Acta. 95: 61-66 (2012) https://doi.org/10.1002/hlca.201100282
  10. Freeman TL, Tuma DJ, Thiele GM, Klassen LW, Worrall S, Niemela O, Parkkila S, Emery PW, Preedy VR. Recent advances in alcohol- induced adduct formation. Alcohol. Clin. Exp. Res. 29: 1310- 1316 (2005) https://doi.org/10.1097/01.ALC.0000171484.52201.52
  11. Gemma S, Vichi S, Testai E. Individual susceptibility and alcohol effects: biochemical and genetic aspects. Ann. Ist. Super. Sanita. 42: 8-16 (2006)
  12. Gomez-Lechon MJ, Donato MT, Martinez-Romero A, Jimenez N, Castell JV, O'Connor J. A human hepatocellular in vitro model to investigate steatosis. Chem. Biol. Interact. 165: 106-116 (2007) https://doi.org/10.1016/j.cbi.2006.11.004
  13. Ha JS, Park SK, Park CH, Seung TW, Guo TJ, Kang JY, Lee DS, Kim JM, Lee U, Heo HJ. Neuronal cell protective effect of new green extract against $H_2O_2$-induced oxidative stress and analysis of bioactive compounds. Korean J. Food Sci. Technol. 47: 673- 679 (2015) https://doi.org/10.9721/KJFST.2015.47.5.673
  14. Han EK, Jin YX, Yoo YS, Jung EJ, Lee JY, Chung CK. Effect of Artemisia capillaris and Paecilomyces japonica on the reduction of hepatotoxicity and lipid metabolism induced by ethanol. J. Korean Soc. Food Sci. Nutr. 38: 1016-1023 (2009) https://doi.org/10.3746/jkfn.2009.38.8.1016
  15. Hwang KA, Hwang YJ, Kim GR, Choe JS. Extracts from Aralia elata (Miq) Seem alleviate hepatosteatosis via improving hepatic insulin sensitivity. BMC complement. Altern. Med. 15: 347 (2015)
  16. Kim MJ, Cho SY, Shin KH, Lee MK. Effects of Aralia elata water extracts on activities of hepatic oxygen free radical generating and scavenging enzymes in streptozotocin-induced diabetic rats. J. Korean Soc. Food Sci. Nutr. 33: 653-658 (2004) https://doi.org/10.3746/jkfn.2004.33.4.653
  17. Kim OK, Ho JN, Nam DE, Jun WJ, Hwang KT, Kang JE, Chae OS, Lee JM. Hepatoprotective effect of Curdrania tricuspidata extracts against oxidative damage. J. Korean Soc. Food Sci. Nutr. 41: 7-13 (2012) https://doi.org/10.3746/jkfn.2012.41.1.007
  18. Kim JS, Shim SH, Chae S, Han SJ, Kang SS, Son KH, Chang HW, Kim HP, Bae K. Saponins and other constituents from the leaves of Aralia elata. Chem. Pharm. Bull. 53: 696-700 (2005) https://doi.org/10.1248/cpb.53.696
  19. Lee JH, Jeong CS. Suppressive effects on the biosynthesis of inflammatory mediators by Aralia elata extract fractions in macrophage cells. Environ. Toxicol. Pharmacol. 28: 333-341 (2009) https://doi.org/10.1016/j.etap.2009.05.009
  20. Lieber CS. Role of oxidative stress and antioxidant therapy in alcoholic and nonalcoholic liver diseases. Adv. Pharmacol. 38: 601- 628 (1996)
  21. Lieber CS. Ethanol metabolism, cirrhosis and alcoholism. Clin. Chim. Acta. 257: 59-84 (1997) https://doi.org/10.1016/S0009-8981(96)06434-0
  22. Lumeng L, Crabb DW. Alcoholic liver disease. Curr. Opin. Gastroenterol. 16: 208-218 (2000) https://doi.org/10.1097/00001574-200005000-00003
  23. Mandrekar P, Szabo G. Signalling pathways in alcohol-induced liver inflammation. J. Hepatol. 50: 1258-1266 (2009) https://doi.org/10.1016/j.jhep.2009.03.007
  24. Nakajima Y, Shimazawa M, Mishima S, Hara H. Water extract of propolis and its main constituents, caffeoylquinic acid derivatives, exert neuroprotective effects via antioxidant actions. Life Sci. 80: 370-377 (2007) https://doi.org/10.1016/j.lfs.2006.09.017
  25. Nhiem NX, Lim HY, Van Kiem P, Van Minh C, Thu VK, Tai BH, Quang TH, Song SB, Kim YH. Oleanane-type triterpene saponins from the bark of Aralia elata and their NF-eB inhibition and PPAR activation signal pathway. Bioorg. Med. Chem. Lett. 21: 6143-6147 (2011) https://doi.org/10.1016/j.bmcl.2011.08.024
  26. Okamoto T, Kanemoto N, Ban T, Sudo T, Nagano K, Niki I. Establishment and characterization of a novel method for evaluating gluconeogenesis using hepatic cell lines, H4IIE and HepG2. Arch. Biochem. Biophys. 491: 46-52 (2009) https://doi.org/10.1016/j.abb.2009.09.015
  27. Olthof MR, Hollman PC, Katan MB. Chlorogenic acid and caffeic acid are absorbed in humans. J. Nutr. 131: 66-71 (2001) https://doi.org/10.1093/jn/131.1.66
  28. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic. Biol. Med. 26: 1231-1237 (1999) https://doi.org/10.1016/S0891-5849(98)00315-3
  29. Sekiya M, Hiraishi A, Touyama M, Sakamoto K. Oxidative stress induced lipid accumulation via SREBP1c activation in HepG2 cells. Biochem. Biophys. Res. Commun. 375: 602-607 (2008) https://doi.org/10.1016/j.bbrc.2008.08.068
  30. Shi H, Dong L, Jiang J, Zhao J, Zhao G, Dang X, Lu X, Jia M. Chlorogenic acid reduces liver inflammation and fibrosis through inhibition of toll-like receptor 4 signaling pathway. Toxicology. 303: 107-114 (2013) https://doi.org/10.1016/j.tox.2012.10.025
  31. Shin KH, Cho SY, Lee MK, Lee JS, Kim MJ. Effects of Aralia elata, Acanthopanacis cortex and Ulmus davidiana water extracts on plasma biomarkers in streptozotocin-induced diabetic rats. J. Korean Soc. Food Sci. Nutr. 33: 1457-1462 (2004) https://doi.org/10.3746/jkfn.2004.33.9.1457
  32. Singleton VL, Rossi JA. Colorimetry of total phenolics with phosphomolybdic- phosphotungstic acid reagents. Am. J. Enol. Vitic. 16: 144-158 (1965)
  33. Urquiaga I, Leighton F. Plant polyphenol antioxidants and oxidative stress. Biol. Res. 33: 55-64 (2000)
  34. Walsh K, Alexander G. Alcoholic liver disease. Postgrad. Med. J. 76: 280-286 (2000) https://doi.org/10.1136/pmj.76.895.280
  35. Yang JW, Kwak CS. Inhibitory effect of Aralia elata ethanol extract against skin damage in UVB-exposed human keratinocytes and human dermal fibroblasts. J. Nutr. Health. 49: 429-436 (2016) https://doi.org/10.4163/jnh.2016.49.6.429
  36. Yoshikawa M, Murakami T, Harada E, Murakami N, Yamahara J, Matsuda H. Bioactive saponins and glycosides. VII. on the hypoglycemic principles from the root cortex of Aralia elata seem.: structure related hypoglycemic activity of oleanolic acid oligoglycoside. Chem. Pharm. Bull. 44: 1923-1927 (1996) https://doi.org/10.1248/cpb.44.1923
  37. You YH, Jung KY, Lee YH, Jun WJ, Lee BY. Hepatoprotective effects of Hovenia Dulcis fruit on ethanol-induced liver damage in vitro and in vivo. J. Korean Soc. Food Sci. Nutr. 38: 154-159 (2009) https://doi.org/10.3746/jkfn.2009.38.2.154