청간해주탕(淸肝解酒湯)의 항산화 작용에 관한 실험적 연구

Experimental Study of Chungganhaeju-tang (Qingganjiejiu-tang) on Oxidative Stress

  • 이지은 (경희대학교 한의과대학 간계내과학교실) ;
  • 이장훈 (경희대학교 한의과대학 간계내과학교실)
  • Lee, Ji-Eun (Dept. of Internal Medicine, College of Oriental Medicine, Kyung-Hee University) ;
  • Lee, Jang-Hoon (Dept. of Internal Medicine, College of Oriental Medicine, Kyung-Hee University)
  • 발행 : 2011.06.30

초록

Objectives : Oxidative stress seems to play a major role in mechanisms by which ethanol causes liver injury. Previous studies have shown that treatment with Chungganhaeju-tang (Qingganjiejiu-tang, CGHJT) has protective effects on alcoholic liver disease. The aim of this study was to investigate the effects of Chungganhaeju-tang on oxidative stress. Materials and Methods : In vitro, we evaluated the inhibitory activities of CGHJT on DPPH (1,1-diphenyl-2-picryl-hydrazyl), xanthine oxidase, trypsin, and hyaluronidase, and measured cell viability, and proliferation. In the cell culture model, we measured the activities of superoxide dismutase (SOD), and catalase (CAT) after CGHJT treatment in C34 and E47 cell lines, HepG2 cells transfected with/without the cytochrome P450 2E1 (CYP2E1) gene. In vivo, we measured malondialdehyde levels in the liver tissue and alcohol concentration in the blood. Results : CGHJT showed significant free radical scavenging activity against DPPH and xanthine oxidase in the in vitro study, and increased cell viability, proliferation, and activities of superoxide dismutase, catalase in C34 and in E47 cell lines. CGHJT reduced malondialdehyde levels and blood alcohol concentration in vivo, as well. Conclusions : This study suggests that CGHJT has antioxidant effects on oxidative stress by reducing lipid peroxidation and inhibiting the ethanol induced suppression of antioxidant enzyme activities.

키워드

참고문헌

  1. 통계청 홈페이지. http://kosis.kr/gen_etl/start.jsp? orgId=350&tblId=TX_35001_A061&conn_path=I2&path=.
  2. Tuma DJ, Casey CA. Dangerous by-products of alcohol breakdown-focus on adducts. Alcohol Res Health 2003;27:285-90.
  3. Castillo T, Koop DR, Kamimura S, Triadafilopoulos G, Tsukamoto H. Role of cytochrome P-450 2E1 in ethanol-,carbon tetrachloride and irondependent microsomal lipid peroxidation. Hepatology 1992;16:992-6. https://doi.org/10.1002/hep.1840160423
  4. 곽미애, 이장훈, 우홍정. 淸肝解酒湯이 알코올대사 및 손상간에 미치는 영향. 대한한의학회지 2000;21(1):68-76.
  5. 이장훈, 박신명, 김영철, 우홍정. 淸肝解酒湯이 알코올성 지방간에 미치는 영향에 대한 임상적 연구. 대한한의학회지 2001;22(4):107-13.
  6. 김영태, 김영철, 이장훈, 우홍정. 淸肝解酒湯이 알코올 대사관련 유전자 및 apoptosis에 미치는 영향. 대한한방내과학회지 2003;24(1):123-33.
  7. 김병삼, 김영철, 이장훈, 우홍정. 淸肝解酒湯이 에탄올 매개성 cytokine발현에 미치는 영향. 대한한의학회지 2003;24(1):190-201.
  8. 윤여광, 김영철, 이장훈, 우홍정. 淸肝解酒湯이 인체간세포의 Glutathione 생성에 미치는 영향. 대한한방내과학회지 2004;25(1):81-91.
  9. 한창우, 김영철, 이장훈, 우홍정. 淸肝解酒湯이 kupffer cell의 NF-$\kappa$B 활성화 및 세포사멸에 미치는 영향. 대한한방내과학회지 2004;25(1):59-70.
  10. 이지현, 김영철, 이장훈, 우홍정. 淸肝解酒湯이 TGF-$\beta1$ 유도성 간섬유화에 미치는 영향. 대한 한방내과학회지 2005;26(1):93-106.
  11. 이지은, 김영철, 이장훈, 우홍정. 淸肝解酒湯이 CYP2E1-transfected HepG2 cell에서 알코올유발 세포독성에 미치는 영향. 대한한방내과학회지 2006;27(1):27-39.
  12. 정윤종, 김영철, 이장훈, 우홍정. 淸肝解酒湯이 알코올성 간손상 Proteome에 미치는 영향. 대한한방내과학회지 2007;28(1):68-78.
  13. 전재현, 김영철, 이장훈, 우홍정. 淸肝解酒湯이 알코올 유발 간섬유화와 단백질 발현에 미치는 영향. 대한한방내과학회지 2008;29(2):469-89.
  14. Arthur I. Cederbaum, Yong Lu, Defeng Wu. Role of oxidative stress in alcohol-induced liver injury. Arch Toxicol 2009;83:519-48. https://doi.org/10.1007/s00204-009-0432-0
  15. 지형준. 대한약전 및 대한약전외 한약규격주해. 서울: 한국메디칼인덱스사; 1998, p. 58, 65, 274, 286, 310, 501, 522, 536, 566, 610, 700.
  16. Blois MS. Antioxidant determinations by the use of a stable free radical. Nature 1958;181:1199-200. https://doi.org/10.1038/1811199a0
  17. Gutfinger T. Polyphenols in olive oils. J Am Oil Chem Soc 1981;58:966-8. https://doi.org/10.1007/BF02659771
  18. Anson ML. The estimation of pepsin, trypsin, papain and cathepsin with hemoglobin. J Gen Physiol 1938;22:79-89. https://doi.org/10.1085/jgp.22.1.79
  19. Uchiyama T, Kamikawa H, Xen-ichi O. Antiinflammatory effect of extract from Phellodendri cortex. J Med Pharm Soc WAKAN-YAKU 1989;6:158-64.
  20. Kim YS, Noh YK, Lee GI, Kim YK, KS, Min KR. Inhibitory effects of herbal medicines on hyaluronidase activity. Kor J Pharmcogn 1995;26(3):265-72.
  21. McCord JM, Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein(hemocuprein). J Biol Chem 1969;244:6049-55.
  22. Johansson LH, Borg LA. A spectrophotometric method for determination of catalase activity in small tissue samples. Anal Biochem 1988;174(1):331-6. https://doi.org/10.1016/0003-2697(88)90554-4
  23. Naik SR, Panda VS. Antioxidant and hepatoprotective effects of Ginkgo biloba phytosomes in carbon tetrachloride-induced liver injury in rodents. Liver Int 2007;27(3):393-9. https://doi.org/10.1111/j.1478-3231.2007.01463.x
  24. Cho MH, Shim SM, Lee SR, Mar WC, Kim GH. Effect of Evodiae fructus extracts on gene expressions related with alcohol metabolism and antioxidation in ethanol-loaded mice. Food Chem Toxicol 2005;43(9):1365-71. https://doi.org/10.1016/j.fct.2005.03.010
  25. Guengerich FP, Shimada T. Oxidation of toxic and carcinogenic chemicaIs by human cytochrorne P-450 enzymes. Chem Res Toxicol 1991;4:391-407. https://doi.org/10.1021/tx00022a001
  26. Lieber CS, Barasona E, Hernandez R, Kubota S, Sato N, Kawano S, et al. Impaired oxyzen utilization, a new metabolism for the hepatotoxicity of ethanol in sub-human primates. J Clin Invest 1989;83:1682-90. https://doi.org/10.1172/JCI114068
  27. 알코올 간질환. 채희복. 대한소화기학회지 2009;53:275-82.
  28. Cederbaum AI, Lu Y, Wu D. Role of oxidative stress in alcohol-induced liver injury. Arch Toxicol 2009;83:519-48. https://doi.org/10.1007/s00204-009-0432-0
  29. Rouach H, Fataccioli V, Gentil M, French SW, Morimoto M, Nordmann R. Effect of chronic ethanol feeding on lipid peroxidation and protein oxidation in relation to liver pathology. Hepatology 1997;25:351-5. https://doi.org/10.1002/hep.510250216
  30. Kessova I, Cederbaum AI. CYP2E1: biochemistry, toxicology, regulation and function in ethanol -induced liver injury. Curr Mol Med 2003;3:509-18. https://doi.org/10.2174/1566524033479609
  31. Morimoto M, Hagbjork AL, Wan YJ, Fu PC, Clot P, Albano E, et al. Modulation of experimental alcohol-induced liver disease by cytochrome P450 2E1 inhibitors. Hepatology 1995;21:1610-7.
  32. Bradford BU, Kona H, Isayama F, Kosyk O, Wheeler MD, Akiyama TE, et al. Cytochrome P450 CYP2E1, but not nicotinamide adenine dinucleotide phosphate oxidase is required for ethanol-induced oxidative DNA damage in rodent liver. Hepatology 2005;41:336-44. https://doi.org/10.1002/hep.20532
  33. Cassidy DA, Mcleod R, Clarke DJ. Regulation of hepatic xanthine oxidoreductase expression by treatment of rats with xenobiotic agents. Biochem Soc Trans 1997;25(4):S563.
  34. 김은영, 백인희, 김정현, 김성란, 유미라. 항산화활성을 나타내는 약용식물 소재 탐색. 한국식품과학회지 2004;36(2):333-8.
  35. 문지숙, 김선재, 박윤미, 황인식, 김의형, 박정욱 등. 약용식물 추출물에 대한 항미생물 활성 검색과 폴리페놀 함량. 한국식품저장유통학회지 2004;11(2):207-13.
  36. McCord JM. Free radicals and inflammation: protection of synorial fluid by superoxide dismutase. Science 1974;185:529-31. https://doi.org/10.1126/science.185.4150.529
  37. Nakagawa H, Watanabe K, Shuto K. Antiinflammatory effect of proteinase inhibitors on carrageenin-induced inflammation in rats. Biochem Pharmacol 1983;32:1191-5. https://doi.org/10.1016/0006-2952(83)90270-8
  38. 문태철, 정광원, 정규찬, 손건호, 김현표, 강삼 식, 등. 천연물로부터 염증성 포스포리파제 A2 저해제 검색. 약학회지 1997;41(5):565-70.
  39. 박종훈, 최혁재, 정석희, 김남재, 김동현. 빈용 한약재의 진통 소염 활성. 생약학회지 2001;32(4):257-68.
  40. 최수임, 이윤미, 허태련. 생약재 추출물의 hyaluronidase 저해 및 라디칼 소거 활성 검색. 한국생물공학회지 2003;18(4):282-8.
  41. Korycka-Dahl M, Richardson T, Hicks C. Superoxide Dismutase Activity in Bovine Milk Serum, J. Food Protection 1979;42:867-71.
  42. Baudrimont I, Ahouandjivo R, Creppy EE. Prevention of lipid peroxidation induced by ochratoxin A in vero cells in culture by several agents. Chem Biol Interact 1997;104:29-40. https://doi.org/10.1016/S0009-2797(97)03764-2
  43. Dianzani MU. Lipid peroxidation in ethanol poisoning: a critical reconsideration. Alcohol 1985;20:161-73.
  44. Recknagel RO. Carbon tetrachoride hepatotoxicity. Pharmacol Rev 1967;19:145-208.
  45. Lieber CS, Barasona E, Hernandez R, Kubota S, Sato N, Kawano S, et al. Impaired oxyzen utilization, a new metabolism for the hepatotoxicity of ethanol in sub-human primates. J Clin Invest 1984;83:1682-90.