Food Engineering Progress (산업식품공학)

Korean Society for Industrial Food Engineering (한국산업식품공학회)
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Effect of Semisulcospira libertina Extracts from Different Extraction Processes on Liver Cell Toxicity and Ethanol Metabolism

간세포 독성과 에탄올 대사에서 추출 조건에 따른 다슬기 추출물의 효과

  • Cho, Kyoung Hwan (Institute of Hadong Green Tea) ;
  • Choo, Ho Jin (Jeongok Co., Ltd.) ;
  • Seo, Min Gyun (Jeongok Co., Ltd.) ;
  • Kim, Jong Cheol (Institute of Hadong Green Tea) ;
  • Shin, Yu Jin (Institute of Hadong Green Tea) ;
  • Ryu, Gi Hyung (Department of Food Science and Technology, Kongju National University) ;
  • Cho, Hee Young (Department of Surgery, Gyeongsang National University School of Medicine, Institute of Health Sciences, Biomedical Research Institute, Gyeongsang National University Hospital) ;
  • Jeong, Chi-Young (Department of Surgery, Gyeongsang National University School of Medicine, Institute of Health Sciences, Biomedical Research Institute, Gyeongsang National University Hospital) ;
  • Hah, Young-Sool (Department of Surgery, Gyeongsang National University School of Medicine, Institute of Health Sciences, Biomedical Research Institute, Gyeongsang National University Hospital)
  • 조경환 ((재)하농녹차연구소) ;
  • 추호진 ((주)정옥) ;
  • 서민균 ((주)정옥) ;
  • 김종철 ((재)하농녹차연구소) ;
  • 신유진 ((재)하농녹차연구소) ;
  • 류기형 (공주대학교 식품공학과) ;
  • 조희영 (경상대학교 의과대학 외과학교실, 건강과학연구원, 경상대학교병원 의생명연구원) ;
  • 정치영 (경상대학교 의과대학 외과학교실, 건강과학연구원, 경상대학교병원 의생명연구원) ;
  • 하영술 (경상대학교 의과대학 외과학교실, 건강과학연구원, 경상대학교병원 의생명연구원)
  • Received : 2017.04.24
  • Accepted : 2017.05.19
  • Published : 2017.05.31
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Abstract

Although Semisulcospira libertina is generally regarded as a supplement for the alleviation of alcohol hangover, little is known about its effects on cell metabolism. Therefore, this study was conducted to analyze the constituents of the extracts prepared using different extraction methods and to compare their biochemical properties. The amino acid contents were found to be much higher in acidic and enzymatic hydrolysates than hot water extracts from S. libertina. DPPH radical scavenging activities in acidic and enzymatic hydrolysates were higher than those of hot water extracts. Three types of S. libertina hydrolysate was added to HepG2 cells damaged by acetaminophen (AAP), after which the survival rate of HepG2 cell were measured. In addition, lactate dehydrogenase (LDH) activities in the culture media were evaluated. The survival rates of HepG2 cells were $77.0{\pm}4.3%$ and $81.5{\pm}1.3%$ at 3 h and 5h enzymatic hydrolysates, respectively. These cell survival rates were higher compared to those of the negative control group ($67.8{\pm}4.3%$) treated only with acetaminophen. Cellular toxicities induced by treatment with AAP were also significantly alleviated in response to treatment with the extracts of S. libertina. In addition, the activities of 2 key enzymes that metabolize ethanol, alcohol dehydrogenase and aldehyde dehydrogenase, were upregulated by 4.7- and 2.7-fold respectively in response to treatment with a 3 h enzymatic hydrolysate of S. libertina. Taken together, these results provide biochemical evidence of the method by which S. libertina exerts its biological functions, including the alleviation of alcohol hangover and the protection of liver cells against toxic insults.

본 연구에서는 다슬기 추출물의 간세포 독성과 에탄올 대사에서의 효능을 분석하기 위하여 산과 효소를 이용하여 추출한 다슬기 가수분해물의 아미노산 조성, 항산화 활성, 아세트아미노펜 처리에 의해 유도된 간독성 대한 보호효과 및 ADH와 ALDH 효소 활성에 미치는 영향을 in vitro에서 분석하였고, 다슬기 추출물을 알코올과 함께 백서에 투여하였을 때 나타나는 혈중 독성 중간대사산물인 아세트알데히드를 정량하여 알코올 섭취 후 나타나는 숙취해소에 대한 효능을 규명하고자 하였다. 열수추출물과 비교하여 산추출물, 효소추출물, 효소-산추출물 모두 총 유리아미노산 함량이 증가하였으며, 특히 숙취해소를 도와주는 아스파라긴산, 글루탐산, 메티오닌 등이 대폭 증가하여 다슬기 추출물의 기능적 향상을 가져올 것으로 생각되었다. 또한, 항산화능의 평가지표인 DPPH 라디칼 소거활성 또한 증가하는 경향을 확인하였다. 다슬기 추출물은 AAP로 간 손상을 유도한 세포에 처리하여 세포생존율과 세포독성을 측정하였다. AAP 만을 처리한 대조군은 $67.8{\pm}4.3%$의 생존율을 보인 반면 다슬기추출물 1 mg/mL과 AAP를 병용처리한 경우 각각 9.7%와 13.7%의 간세포 생존율 증가를 보였다. 세포독성은 AAP 만을 처리한 대조군은 33.7%의 LDH 활성에 의한 세포독성을 나타낸 반면 다슬기 추출물을 처리한 경우 대조군에 비해 세포독성이 15.4%-24.4%로 감소되었다. 또한, 에탄올 대사에서 중요한 ADH와 ALDH 활성은 열수추출물에 비해 효소를 이용한 다슬기 추출물에서 각각 최대 4.8배와 2.7배 증가하였다. 알코올 섭취 후 혈중 아세트알데히드 농도는 다슬기 효소 추출물을 투여한 군의 경우 양성대조군에 비하여 유의하게 감소하였다. 이상의 연구결과를 종합해 보면 다양한 추출 방법에 의한 다슬기추출물 소재는 항산화 활성과 간손상 보호효과 및 숙취해소능을 가지며, 효소추출물의 ADH와 ALDH 활성 증가효과가 더 우수하였으며 향후 이들 소재를 활용한 다양한 기능성 제품의 개발이 필요할 것으로 판단된다.

Keywords

Acknowledgement

Supported by : 산업통상자원부

References

  1. Blois MS. 1958. Antioxidant determinations by the use of a stable free radical. Nature 181: 1199-1200. https://doi.org/10.1038/1811199a0
  2. Cha HO, Baik WA. 1999. Effect of Semisulcospira libertina, Corbicula sp. and Cipangopaludina chinensis malleata extracts on liver function and other biological activities. 45th National Sci. Exhibition of Ministry of Science and Technology, Sejong, Korea
  3. Cha JY, Jung HJ, Jeong JJ, Yang HJ, Kim YT, Lee YS. 2009. Effects of amino acids on the activities of alcohol metabolizing enzyme alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). J. Life Sci. 19: 1321-1327. https://doi.org/10.5352/JLS.2009.19.9.1321
  4. Grunnet N, Kondrup J. 1986. The effect of ethanol on the betaoxidation of fatty acids. Alcohol. Clin. Exp. Res. 10: 64S-68S. https://doi.org/10.1111/j.1530-0277.1986.tb05182.x
  5. Di Luzio N, Hartman A. 1967. Role of lipid peroxidation in the pathogenesis of the ethanol-induced fatty liver. Fed. Proc. 26: 1436-1442.
  6. Eriksson CJ. 2001. The role of acetaldehyde in the actions of alcohol. Alcohol. Clin. Exp. Res. 25: 15S-32S. https://doi.org/10.1111/j.1530-0277.2001.tb02369.x
  7. Forsander OA, Raiha NC. 1960. Metabolites produced in the liver during alcohol oxidation. J. Biol. Chem. 235: 34-36.
  8. Glueck CJ, HoggE, AllenC, Gartside PS. 1980. Effects of alcohol ingestion on lipid and lipoprotein in normal man. Am. J. Clin. Nutr. 33: 2287-2293. https://doi.org/10.1093/ajcn/33.11.2287
  9. Heineckem JW. 1987. Free radical modification of low-density lipoprotein: mechanisms and biological consequences. Free Rad. Biol. Med. 3: 65-73. https://doi.org/10.1016/0891-5849(87)90040-2
  10. Hwang JY, Ham JW, Nam SH. 2004. Effect of maesil (Prunus mume) juice on the alcohol metabolizing enzyme activities. Korean J. Food Sci. Technol. 36: 329-332.
  11. Hwang SJ, Choi HM, Park HJ, Lee JS, Heo D, Kim MR. 2010. Effects of medicinal herbal drink on alcohol metabolic enzyme in drunken rats. J. Physiol. Pathol. Korean Med. 24: 610-615
  12. Iimuro Y, Bradford BU, Forman DT, Thurman RG. 1996. Glycine prevents alcohol-induced liver injury by decreasing alcohol in the rat stomach. Gastroenterology 110: 1536-1542. https://doi.org/10.1053/gast.1996.v110.pm8613061
  13. Jeon TW, Lee YS, Kim HJ. 2003. Hepatoprotection by Semisulcospira libertina against acetaminophen-induced hepatic injury in mice. J. Food Sci. Nutr. 8: 239-244.
  14. Kim YK, Moon HS, Lee, MH, Park MJ, Lim CW, Paek HY, Park JI, Yoon HD, Kim DH. 2009. Biological activities of seven melania snails in Korea. Kor. J. Fish Aquat. Sci. 42: 434-441.
  15. Lee MH, Kim YK, Moon HS, Kim YA, Yoon NY, Lim CW, Park HY, Kim DH. 2010. Antioxidant activity of five melania snails of the genus Semisulcospira in Korea. Kor. J. Fish Aquat. Sci. 43: 188-194.
  16. Lee MS, Park JB, Yoon SH. 2005. Hepatoprotective effect of the water extract from Semisulcospira gottschei against liver injuries induced by carbon tetrachloride in rats. J. Korean Soc. Hyg. Sci. 11: 17-26.
  17. Lieber CS. 1994. Alcohol and the liver: 1994 update. Gastroenterology 106: 1085-1090. https://doi.org/10.1016/0016-5085(94)90772-2
  18. Lieber CS. 1997. Ethanol metabolism, cirrhosis and alcoholism. Clin. Chim. Acta 257: 59-84. https://doi.org/10.1016/S0009-8981(96)06434-0
  19. Lim CW, Kim YK, Kim DH, Park JI, Lee MH, Park HY, Jang MS. 2009. Comparison of quality characteristics of melania snails in Korea. Kor. J. Fish Aquat. Sci. 42: 555-560.
  20. Lin RC, Li TK. 1998. Effets of isoflavones on alcohol pharmacokinetics and alcohol-drinking behavior in rat. Am. J. Clin. Nutr. 68: 1512S-1515S. https://doi.org/10.1093/ajcn/68.6.1512S
  21. Linder MC. 1991. Nutrition and metabolism of fats. In: Nutritional Biochemistry and Metabolism: with Clinical Applications. 2nd ed. Elsevier, Amsterdam, Oxford, NY, USA. pp 79-83.
  22. Magonet E, Hayen P, Delforge D, Delaive E, Remacle J. 1992. Importance of the structural zinc atom for the stability of yeast alcohol dehydrogenase. Biochem. J. 287: 361-365. https://doi.org/10.1042/bj2870361
  23. Mezey E. 1980. Alcoholic liver disease; roles of alcohol and malnutrition. Am. J. Clin. Nutr. 33: 2709-2718. https://doi.org/10.1093/ajcn/33.12.2709
  24. Morel DW, DiCorleto PE, Chisolm GM. 1984. Endothelial and smooth muscle cells after low density lipoproteins in vitro by free radical oxidation. Arteriosclerosis, 4: 357-364. https://doi.org/10.1161/01.ATV.4.4.357
  25. Park SC. 1993. Ethanol oxidation is accelerated by augmentation of malate-aspartate shuttle with aspartate. Korean J. Biochem. 25: 137-143.
  26. Re R, Pellegrini N, Protegente 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
  27. Shen L, Hillebrand A, Wang DQ, Liu M. 2012. Isolation and primary culture of rat hepatic cells. J. Vis. Exp. 29. pii: 3917.1
  28. Wu D, Cederbaum AI. 1999. Ethanol cytotoxicity to a transfected HepG2 cell line expressing human cytochrome P450E1. Alcohol. Clin. Exp. Res. 23: 67-76. https://doi.org/10.1111/j.1530-0277.1999.tb04025.x
  29. Yin M, Ikejima K, Arteet GE, Seabra V, Bradford BU, Kono H, Rusyn I, Thurman RG. 1998. Glycine accelerates recovery from alcohol-induced liver injury. J. Pharacol. Exp. Ther. 286: 1014-1019.

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