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

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Hepatoprotective Effects of Oyster Hydrolysate on Lipopolysaccharide/D-Galactosamine-Induced Acute Liver Injury in Mice

Lipopolysaccharide/D-Galactosamine에 의해 유도된 급성 간 손상 생쥐모델에서 굴가수분해물의 간 보호 효과

  • Ryu, Ji Hyeon (Department of Physiology, College of Medicine, Gyeongsang National University) ;
  • Kim, Eun-Jin (Department of Physiology, College of Medicine, Gyeongsang National University) ;
  • Xie, Chengliang (Department of Anatomy, College of Medicine, Gyeongsang National University) ;
  • Nyiramana, Marie Merci (Department of Physiology, College of Medicine, Gyeongsang National University) ;
  • Siregar, Adrian S. (Department of Physiology, College of Medicine, Gyeongsang National University) ;
  • Park, Si-Hyang (Sun Marine Biotech Co.) ;
  • Cho, Soo Buem (Department of Radiology, Gyeongsang National University Changwon Hospital) ;
  • Song, Dae Hyun (Institute of Health Sciences, College of Medicine, Gyeongsang National University) ;
  • Kim, Nam-Gil (Department of Marine Biology and Aquaculture and Institute of Marine Industry, Gyeongsang National University) ;
  • Choi, Yeung Joon (Department of Seafood Science and Technology and Institute of Marine Industry, Gyeongsang National University) ;
  • Kang, Sang Soo (Institute of Health Sciences, College of Medicine, Gyeongsang National University) ;
  • Kang, Dawon (Department of Physiology, College of Medicine, Gyeongsang National University)
  • 류지현 (경상대학교 의과대학 생리학교실) ;
  • 김은진 (경상대학교 의과대학 생리학교실) ;
  • ;
  • ;
  • ;
  • 박시향 (선마린바이오테크) ;
  • 조수범 (경상대학교 창원병원 영상의학과) ;
  • 송대현 (경상대학교 의과대학 건강과학연구원) ;
  • 김남길 (경상대학교 해양과학대학 해양생명과학과) ;
  • 최영준 (경상대학교 해양과학대학 해양식품공학과) ;
  • 강상수 (경상대학교 의과대학 건강과학연구원) ;
  • 강다원 (경상대학교 의과대학 생리학교실)
  • Received : 2017.03.29
  • Accepted : 2017.05.19
  • Published : 2017.06.30
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Abstract

Oxidative stress and inflammation are key factors responsible for progression of liver injury. A variety of functions of oyster hydrolysate (OH) are affected by their antioxidant and anti-inflammatory activities. However, little is known regarding the effects of OH on a liver injury model. This study was performed to evaluate the effects of OH on acute liver injury induced by lipopolysaccharide/D-galactosamine (LPS/D-GalN) in mice. Experimental groups were divided into six groups as follows (each group, n=10): control (saline), LPS/D-GalN, LPS/D-GalN+OH (100 mg/kg), LPS/D-GalN+OH (200 mg/kg), LPS/D-GalN+OH (400 mg/kg), and LPS/D-GalN+silymarin (25 mg/kg, positive control). The experimental acute liver injury model was induced with LPS ($1{\mu}g/kg$) and D-GalN (400 mg/kg). We first analyzed antioxidant and anti-inflammatory activities in OH. OH showed high DPPH and ABTS radical scavenging activities and reduced ROS generation in Chang cells in a dose-dependent manner. In addition, OH showed anti-inflammatory activities, such as inhibition of cyclooxygenase-2 and 5-lipooxygenase. Treatment with OH down-regulated tumor necrosis factor $(TNF)-{\alpha}$, interleukin (IL)-6, and $IL-1{\alpha}$ expression levels in LPS-stimulated RAW264.7 cells. OH significantly reduced LPS/D-GalN-induced increases in the concentrations of alanine transaminase and aspartate aminotransferase in serum. In the LPS/D-GalN group, liver tissues exhibited apoptosis of hepatocytes with hemorrhages. These pathological alterations were ameliorated by OH treatment. Consistently, hepatic catalase activity was low in the LPS/D-GalN group compared to the control group, and catalase activity was significantly restored by OH treatment (P<0.05). Furthermore, OH markedly reduced the LPS/D-GalN-induced increase in $TNF-{\alpha}$, $IL-1{\beta}$, and IL-6 levels in liver tissue. Taken together, these results show that OH has hepatoprotective effects on LPS/D-GalN-induced acute liver injury via inhibition of oxidative stress and inflammation, suggesting that OH could be used as a health functional food and potential therapeutic agent for acute liver injury.

산화스트레스와 염증은 간 손상의 진행과정에 중요한 인자로 작용한다. 굴가수분해물의 항산화 및 항염증 활성은 지질대사, 혈압 및 혈당, 면역기능의 조절과 같은 다양한 기능에 관여한다. 그러나 급성 간 손상 모델에서 굴가수분해물의 효과를 확인한 연구 결과는 아직 확인된 바 없다. 본 연구는 LPS/D-GalN에 의해 유도된 급성 간 손상 생쥐 모델에서 굴가수분해물의 효과를 확인하기 위해 수행되었다. 실험군은 대조군(생리식염수), LPS/D-GalN 간 손상군, LPS/D-GalN과 굴가수분해물(100 mg/kg, 200 mg/kg, 400 mg/kg)의 병합투여군 및 LPS/D-GalN과 silymarin(25 mg/kg) 병합투여군으로 나누었다. 급성 간 손상 모델은 $1{\mu}g/kg$의 LPS와 400 mg/kg의 D-GalN으로 유도되었다. 먼저 시료의 항산화 및 항염증 활성을 분석한 결과 굴가수분해물은 농도 의존적으로 높은 DPPH 및 ABTS 라디칼 소거 활성을 보였으며, 인간 정상 간세포주(Chang)에서 과산화수소에 의한 세포 내 활성산소의 생성을 유의적으로 감소시켰다. 또한, 굴가수분해물은 농도 의존적으로 높은 COX-2 및 5-LOX 억제능을 보였으며, LPS에 의해 활성화된 생쥐 대식세포주 RAW264.7에서 발현되는 $TNF-{\alpha}$, IL-6 및 $IL-1{\beta}$의 염증성 사이토카인의 mRNA 발현률을 감소시켰다. 굴가수분해물 투여는 LPS/D-GalN에 의한 혈청 ALT 및 AST 증가를 유의적으로 감소시켰으며, 간 조직의 출혈 및 간세포의 자멸사를 감소시켰다. 또한, 간 균질의 $TNF-{\alpha}$, $IL-1{\beta}$ 및 IL-6 함량을 감소시켰으며, 감소한 catalase의 활성을 유의적으로 증가시켰다. 이상의 결과로부터 굴가수분해물은 간 보호 효과를 가지는 것으로 판단되며, 급성 간 손상의 예방 및 치료에 도움이 될 수 있는 시료로 활용될 수 있을 것으로 기대된다.

Keywords

References

  1. Bernal W, Wendon J. 2013. Acute liver failure. N Engl J Med 369: 2525-2534. https://doi.org/10.1056/NEJMra1208937
  2. Escorsell A, Mas A, de la Mata M. 2007. Acute liver failure in Spain: analysis of 267 cases. Liver Transpl 13: 1389-1395. https://doi.org/10.1002/lt.21119
  3. Bower WA, Johns M, Margolis HS, Williams IT, Bell BP. 2007. Population-based surveillance for acute liver failure. Am J Gastroenterol 102: 2459-2463. https://doi.org/10.1111/j.1572-0241.2007.01388.x
  4. Kumar R, Shalimar, Bhatia V, Khanal S, Sreenivas V, Gupta SD, Panda SK, Acharya SK. 2010. Antituberculosis therapy-induced acute liver failure: magnitude, profile, prognosis, and predictors of outcome. Hepatology 51: 1665-1674. https://doi.org/10.1002/hep.23534
  5. Ichai P, Samuel D. 2008. Etiology and prognosis of fulminant hepatitis in adults. Liver Transpl 14: S67-S79. https://doi.org/10.1002/lt.21612
  6. Lee WM, Squires RH Jr, Nyberg SL, Doo E, Hoofnagle JH. 2008. Acute liver failure: Summary of a workshop. Hepatology 47: 1401-1415.
  7. Galanos C, Freudenberg MA, Reutter W. 1979. Galactosamine-induced sensitization to the lethal effects of endotoxin. Proc Natl Acad Sci U S A 76: 5939-5943. https://doi.org/10.1073/pnas.76.11.5939
  8. Eipel C, Kidess E, Abshagen K, LeMinh K, Menger MD, Burkhardt H, Vollmar B. 2007. Antileukoproteinase protects against hepatic inflammation, but not apoptosis in the response of D-galactosamine-sensitized mice to lipopolysaccharide. Br Pharmacol 151: 406-413.
  9. Neihörster M, Inoue M, Wendel A. 1992. A link between extracellular reactive oxygen and endotoxin-induced release of tumour necrosis factor $\alpha$ in vivo. Biochem Pharmacol 43: 1151-1154.
  10. Mayer AM, Spitzer JA. 1993. Modulation of superoxide anion generation by manoalide, arachidonic acid and staurosporine in liver infiltrated neutrophils in a rat model of endotoxemia. J Pharmacol Exp Ther 267: 400-409.
  11. Yang F, Li X, Wang LK, Wang LW, Han XQ, Zhang H, Gong ZJ. 2014. Inhibitions of $NF-{\kappa}B$ and $TNF-{\alpha}$ result in differential effects in rats with acute on chronic liver failure induced by d-Gal and LPS. Inflammation 37: 848-857. https://doi.org/10.1007/s10753-013-9805-x
  12. Jung SH, Kim SJ, Jun BG, Lee KT, Hong SP, Oh MS, Jang DS, Choi JH. 2013. $\alpha$-Cyperone, isolated from the rhizomes of Cyperus rotundus, inhibits LPS-induced COX-2 expression and PGE2 production through the negative regulation of $NF{\kappa}B$ signalling in RAW 264.7 cells. J Ethnopharmacol 147: 208-214. https://doi.org/10.1016/j.jep.2013.02.034
  13. Streetz K, Leifeld L, Grundmann D, Ramakers J, Eckert K, Spengler U, Brenner D, Manns M, Trautwein C. 2000. Tumor necrosis factor $\alpha$ in the pathogenesis of human and murine fulminant hepatic failure. Gastroenterology 119: 446-460. https://doi.org/10.1053/gast.2000.9364
  14. You TJ. 1993. Source book of food. Seo-Woo, Seoul, Korea. p 168-169.
  15. Chung IK, Kim HS, Kang KT, Choi YJ, Choi JD, Kim JS, Heu MS. 2006. Preparation and functional properties of enzymatic oyster hydrolysates. J Korean Soc Food Sci Nutr 35: 919-925. https://doi.org/10.3746/jkfn.2006.35.7.919
  16. Wang Q, Li W, He Y, Ren D, Kow F, Song L, Yu X. 2014. Novel antioxidative peptides from the protein hydrolysate of oysters (Crassostrea talienwhanensis). Food Chem 145: 991-996. https://doi.org/10.1016/j.foodchem.2013.08.099
  17. Umayaparvathi S, Meenakshi S, Vimalraj V, Arumugam M, Balasubramanian T. 2014. Isolation and structural elucidation of antioxidant peptides from oyster (Saccostrea cucullata) protein hydrolysate. Protein Pept Lett 21: 1073-1083. https://doi.org/10.2174/0929866521666140417121616
  18. Qian ZJ, Jung WK, Byun HG, Kim SK. 2008. Protective effect of an antioxidative peptide purified from gastrointestinal digests of oyster, Crassostrea gigas against free radical induced DNA damage. Bioresour Technol 99: 3365-3371. https://doi.org/10.1016/j.biortech.2007.08.018
  19. Hwang JW, Lee SJ, Kim YS, Kim EK, Ahn CB, Jeon YJ, Moon SH, Jeon BT, Park PJ. 2012. Purification and characterization of a novel peptide with inhibitory effects on colitis induced mice by dextran sulfate sodium from enzymatic hydrolysates of Crassostrea gigas. Fish Shellfish Immunol 33: 993-999. https://doi.org/10.1016/j.fsi.2012.08.017
  20. Cheong SH, Kim EK, Hwang JW, Kim YS, Lee JS, Moon SH, Jeon BT, Park PJ. 2013. Purification of a novel peptide derived from a shellfish, Crassostrea gigas, and evaluation of its anticancer property. J Agric Food Chem 61: 11442-11446. https://doi.org/10.1021/jf4032553
  21. Shiozaki K, Shiozaki M, Masuda J, Yamauchi A, Ohwada S, Nakano T, Yamaguchi T, Saito T, Muramoto K, Sato M. 2010. Identification of oyster-derived hypotensive peptide acting as angiotensin-I-converting enzyme inhibitor. Fish Sci 76: 865-872. https://doi.org/10.1007/s12562-010-0264-0
  22. Xie CL, Kim JS, Ha JM, Choung SY, Choi YJ. 2014. Angiotensin I-converting enzyme inhibitor derived from cross-linked oyster protein. Biomed Res Int 2014: 379234.
  23. Wang J, Hu J, Cui J, Bai X, Du Y, Miyaguchi Y, Lin B. 2008. Purification and identification of a ACE inhibitory peptide from oyster proteins hydrolysate and the antihypertensive effect of hydrolysate in spontaneously hypertensive rats. Food Chem 111: 302-308. https://doi.org/10.1016/j.foodchem.2008.03.059
  24. Hur SI, Park SH, Lee SS, Choung SY, Choi YJ. 2013. Antioxidative effect of oyster hydrolysate on the serum and hepatic homogenate in SD-rats. J Korean Soc Food Sci Nutr 42: 1940-1948. https://doi.org/10.3746/jkfn.2013.42.12.1940
  25. Kim HA, Park SH, Lee SS, Choi YJ. 2015. Anti-wrinkle effects of enzymatic oyster hydrolysate and its fractions on human fibroblasts. J Korean Soc Food Sci Nutr 44: 1645-1652. https://doi.org/10.3746/jkfn.2015.44.11.1645
  26. Park SH, Moon SS, Xie CL, Choung SY, Choi YJ. 2014. Protective effects of enzymatic oyster hydrolysate on acetaminophen-induced HepG-2 cell damage. J Korean Soc Food Sci Nutr 43: 1166-1173. https://doi.org/10.3746/jkfn.2014.43.8.1166
  27. Blois MS. 1958. Antioxidant determinations by the use of a stable free radical. Nature 181: 1199-1200. https://doi.org/10.1038/1811199a0
  28. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med 26: 1231-1237. https://doi.org/10.1016/S0891-5849(98)00315-3
  29. Reddy CM, Bhat VB, Kiranmai G, Reddy MN, Reddanna P, Madyastha KM. 2000. Selective inhibition of cyclooxygenase-2 by C-phycocyanin, a biliprotein from Spirulina platensis. Biochem Biophys Res Commun 277: 599-603. https://doi.org/10.1006/bbrc.2000.3725
  30. Lyckander IM, Malterud KE. 1992. Lipophilic flavonoids from Orthosiphon spicatus as inhibitors of 15-lipoxygenase. Acta Pharm Nord 4: 159-166.
  31. Park HJ, Jeon BT, Kim HC, Roh GS, Shin JH, Sung NJ, Han J, Kang D. 2012. Aged red garlic extract reduces lipopolysaccharide- induced nitric oxide production in RAW 264.7 macrophages and acute pulmonary inflammation through haeme oxygenase-1 induction. Acta Physiol 205: 61-70. https://doi.org/10.1111/j.1748-1716.2012.02425.x
  32. Gough DR, Cotter TG. 2011. Hydrogen peroxide: a Jekyll and Hyde signalling molecule. Cell Death Dis 2: e213. https://doi.org/10.1038/cddis.2011.96
  33. Aebi H. 1984. Catalase in vitro. Methods Enzymol 105: 121-126.
  34. Jeong YY, Ryu JH, Shin JH, Kang MJ, Kang JR, Han J, Kang D. 2016. Comparison of anti-oxidant and anti-inflammatory effects between fresh and aged black garlic extracts. Molecules 21: 430. https://doi.org/10.3390/molecules21040430
  35. Kim SB, Yeum DM, Yeo SG, Ji CI, Lee YW, Park YH. 1989. Antioxidative effects of food protein hydrolysates by protease. Korean J Food Sci Technol 21: 492-497.
  36. Bertolini A, Ottani A, Sandrini M. 2001. Dual acting anti-inflammatory drugs: a reappraisal. Pharmacol Res 44: 437-450. https://doi.org/10.1006/phrs.2001.0872
  37. Santos CMM, Ribeiro D, Silva AMS, Fernandes E. 2017. 2,3-Diarylxanthones as potential inhibitors of arachidonic acid metabolic pathways. Inflammation 40: 956-964. https://doi.org/10.1007/s10753-017-0540-6
  38. Horrillo R, Planaguma A, Gonzalez-Periz A, Ferre N, Titos E, Miquel R, Lopez-Parra M, Masferrer JL, Arroyo V, Claria J. 2007. Comparative protection against liver inflammation and fibrosis by a selective cyclooxygenase-2 inhibitor and a nonredox-type 5-lipoxygenase inhibitor. J Pharmacol Exp Ther 323: 778-786. https://doi.org/10.1124/jpet.107.128264
  39. Yin G, Huang J, Ma M, Suo X, Huang Z. 2016. Oyster crude polysaccharides attenuates lipopolysaccharide-induced cytokines production and $PPAR{\gamma}$ expression in weanling piglets. Springerplus 5: 677. https://doi.org/10.1186/s40064-016-2319-x
  40. Lee SY, Kim HJ, Han JS. 2013. Anti-inflammatory effect of oyster shell extract in LPS-stimulated Raw 264.7 cells. Prev Nutr Food Sci 18: 23-29. https://doi.org/10.3746/pnf.2013.18.1.023
  41. Chung K, Cho SH, Sin EN, Choi KH, Choi YS. 1988. Effects of alcohol consumption and fat content in diet on chemical composition and morphology of liver in rat. Korean J Nutr 21: 154-163.
  42. Reddy MK, Reddy AG, Kumar BK, Madhuri D, Boobalan G, Reddy MA. 2017. Protective effect of rutin in comparison to silymarin against induced hepatotoxicity in rats. Vet World 10: 74-80. https://doi.org/10.14202/vetworld.2017.74-80
  43. Zhou J, Liang Y, Pan JX, Wang FF, Lin XM, Ma RJ, Qu F, Fang JQ. 2015. Protein extracts of Crassostrea gigas alleviate $CCl_4$-induced hepatic fibrosis in rats by reducing the expression of CTGF, $TGF-{\beta}1$ and $NF-{\kappa}B$ in liver tissues. Mol Med Rep 11: 2913-2920. https://doi.org/10.3892/mmr.2014.3019
  44. Shi X, Ma H, Tong C, Qu M, Jin Q, Li W. 2015. Hepatoprotective effect of a polysaccharide from Crassostrea gigas on acute and chronic models of liver injury. Int J Biol Macromol 78: 142-148. https://doi.org/10.1016/j.ijbiomac.2015.03.056
  45. Zhang C, Li X, Jing X, Zhang B, Zhang Q, Niu Q, Wang J, Tian Z. 2014. Protective effects of oyster extract against hepatic tissue injury in alcoholic liver diseases J Ocean Univ China 13: 262-270. https://doi.org/10.1007/s11802-014-2449-0
  46. Bhaduri BR, Mieli-Vergani G. 1996. Fulminant hepatic failure: pediatric aspects. Semin Liver Dis 16: 349-355. https://doi.org/10.1055/s-2007-1007248
  47. Kwak SD, Kim CS, Kang CB, Koh PO, Seo DL, Yang JH. 2000. Apoptosis of livers induced by D-galactosamine and lipopolysaccharide in mice. Korean J Vet Res 40: 213-220.
  48. Osaki K, Shimizu Y, Yamamoto T, Miyake F, Kondo S, Yamaguchi H. 2015. Improvement of liver function by the administration of oyster extract as a dietary supplement to habitual alcohol drinkers: A pilot study. Exp Ther Med 10: 705-710. https://doi.org/10.3892/etm.2015.2563
  49. Park HJ, Do HJ, Kim OJ, Kim A, Ha JM. 2012. Hepatoprotective effects of various enzyme hydrolysates from oysters on tacrine-induced toxicity in human hepatoma cells. J Life Sci 22: 117-125. https://doi.org/10.5352/JLS.2012.22.1.117
  50. Moon PD, Kim MH, Lim HS, Oh HA, Nam SY, Han NR, Kim MJ, Jeong HJ, Kim HM. 2015. Taurine, a major amino acid of oyster, enhances linear bone growth in a mouse model of protein malnutrition. Biofactors 41: 190-197. https://doi.org/10.1002/biof.1213
  51. Wu G, Yang J, Sun C, Luan X, Shi J, Hu J. 2009. Effect of taurine on alcoholic liver disease in rats. Adv Exp Med Biol 643: 313-322.
  52. Chang YY, Chou CH, Chiu CH, Yang KT, Lin YL, Weng WL, Chen YC. 2010. Preventive effects of taurine on development of hepatic steatosis induced by a high-fat/cholesterol dietary habit. J Agric Food Chem 59: 450-457.
  53. Fard JK, Hamzeiy H, Sattari M, Eghbal MA. 2016. Protective roles of N-acetyl cysteine and/or taurine against sumatriptaninduced hepatotoxicity. Adv Pharm Bull 6: 627-637. https://doi.org/10.15171/apb.2016.077
  54. El-Houseini ME, El-Agoza IA, Sakr MM, El-Malky GM. 2017. Novel protective role of curcumin and taurine combination against experimental hepatocarcinogenesis. Exp Ther Med 13: 29-36. https://doi.org/10.3892/etm.2016.3952
  55. Kristinsson HG, Rasco BA. 2000. Fish protein hydrolysates: production, biochemical, and functional properties. Crit Rev Food Sci Nutr 40: 43-81. https://doi.org/10.1080/10408690091189266