The Journal of Internal Korean Medicine (대한한방내과학회지)

The Society of Internal Korean Medicine (대한한방내과학회)
권호 표지

Effect of Yinjinchunggan-tang based Herb Formulae Containing Wasong and Eosungcho on Fibrogenesis

인진청간탕 및 와송 어성초 가미방의 간섬유화억제에 미치는 효과

  • Moon, Young-Hoon (Dept. of Internal Medicine, College of Oriental Medicine, Kyung-Hee University) ;
  • Woo, Hong-Jung (Dept. of Internal Medicine, College of Oriental Medicine, Kyung-Hee University)
  • 문영훈 (경희대학교 한의과대학 간계내과학교실) ;
  • 우홍정 (경희대학교 한의과대학 간계내과학교실)
  • Published : 2011.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Objectives : This study was performed to investigate the anti-fibrogenic effect and the effect on cell growth and apoptosis in YJCGT, YJCGT YSO and YJCGT YSCO on thioacetamide-induced rat liver tissue and the immortalized human hepatic cell line LX2. Materials and Methods : LX2 cells were treated with various concentrations (0, 50, 150, 300 ug/ml) of YJCGT, Y+YSO, and Y+YSCO extract for 24, 48 and 72 hours. After the treatment, cell viability was measured by using MTT assay. Caspase inhibitor assay, and cell viability were determined by a colorimetric assay with PMS/MTS solution. Rat liver fibrosis was induced by intraperitoneal thioacetamide injection 150 mg/kg 3 times a week for 5 weeks. After the treatment, body weight, liver & spleen weights, liver function test, the complete blood cell count and the change of portal pressure were studied. After YJCGT, Y+YSO, and Y+YSCO treatment, percentages of collagen in thioacetamide-induced rat liver tissue were measured. Results : The viability of the LX2 cell decreased in a dose- and time-dependent manner. Exposure of LX2 cells to YJCGT, YJCGT+YSO and YJCGT+YSCO induced caspase-3 activation, but co-treatment of YJCGT, YJCGT+YSO and YJCGT+YSCO with the pan-caspase inhibitor Z-VAD-FMK, and the caspase-3 inhibitor Z-DEVE-FMK, blocked apoptosis. There was no difference in rat body weight between the thioacetamide only group and the YJCGT, YJCGT+YSO and YJCGT+YSCO groups. In the YJCGT, YJCGT YSO and YJCGT YSCO groups, the serum level of GPT significantly went down compared with the thioacetamide only group. In the YJCGT, Y+YSO, Y+YSCO groups, white blood cell elevated by thioacetamide injection decreased but RBC, Hgb, and Hct increased. In the Y+YSO group, the portal pressure elevated by thioacetamide injection significantly decreased. In the histological finding, thioacetamide injections caused severe fibrosis, but YJCGT, Y+YSO, and Y+YSCO treatment significantly reduced the amounts of hepatic collagens. Conclusions : YJCGT, Y+YSO, and Y+YSCO inhibit the growth of LX2 cells by inducing apoptosis through caspase activity. YJCGT, Y+YSO, and Y+YSCO have beneficial effects on the treatment of cirrhotic patients as well as patients with chronic hepatitis.

Keywords

References

  1. 통계청. 2009 사망원인통계
  2. 이관식. 간섬유화. 대한소화기학회지 2006;48:297-305.
  3. 우홍정. 만성B형간염에 대한 인진청간탕의 효과. 제2회 한중 학술대회 참가 논문집(간장병). 1995;18-53.
  4. 김영철, 이장훈, 우홍정. 인진청간탕의 안전성에 관한 연구. 경희한방대논문집 1997;20(1):57-89.
  5. 강경태, 이장훈, 우홍정. 인진청간탕가미방이 실험적 흰쥐의 간경변증에 미치는 영향. 경희한의대논문집 1997;20(2):133-50.
  6. 문영훈. 와송이 HepG2 cell의 세포분열 및 관련 유전자 발현에 미치는 영향. 대한한방내과학회지 2005;26(1):48-59.
  7. 윤경수. 와송이 K562에서 세포사멸에 미치는 영향. 대한한방내과학회지 2006;27(1):166-77.
  8. 안덕균. 원색한국본초도감. 교학사; 1998, p. 69.
  9. 지형준 외. 대한약전 및 대한약전외 한약규격주해. 서울: 한국메디칼인덱스사; 1998.
  10. 전국한의과대학 간계내과학 교수 공저. 간계내과학. 서울: 동양의학연구원; 2001, p. 323-5.
  11. Friedman SL. Liver fibrosis-from bench to beside. J Hepatol 2003;38 supple 1:s38-s53.
  12. Williams EJ, Benyon RC, Trim N, Hadwin R, Grove BH, Arthur MJ et al. Ralaxin inhibits effective deposition by cultured hepatic stellate cells and decreases rat liver fibrosis in vivo. Gut 2001;49(4):577-8. https://doi.org/10.1136/gut.49.4.577
  13. Simile MM, Banni S, Angioni E, Carta G, De Miglio MR, Muroni MR et al. 5'-Methylthioadenosine administration prevents lipid peroxidation and fibrogenesis induced in rat liver by carbontetrachloride intoxication. J Hepatol 2001;34(3) :386-94. https://doi.org/10.1016/S0168-8278(00)00078-7
  14. Yoshiji H, Noguchi R, Kuriyama S, Ikenaka Y, Yoshii J, Yanase K et al. Imatinib mesylate (STI-571) attenuates liver fibrosis development in rats. Am J Physiol 2005;288(5):907-13.
  15. Rockey DC. Antifibrotic therapy in chronic liver disease. Clin Gastroenterol Hepatol 2005;3(2):95-107. https://doi.org/10.1016/S1542-3565(04)00445-8
  16. Breitkopf K, Haas S, Wiercinska E, Singer MV, Dooley S. Anti-TGF-beta strategies for the treatment of chronic liver disease. Alcohol Clin Exp Res. 2005;29(11 supple):121S-131S.
  17. Tangkijvanich P, Yee HF Jr. Cirrhosis--can we reverse hepatic fibrosis? Eur J Surg Suppl 2002;587:100-12.
  18. 이영석. 만성 간질환에서 항섬유화 치료의 과거와 미래. 대한간학회지 2006;12(1s):41-55.
  19. 이관식, 간섬유화 기전. 대한간학회지 2006;11(2s):9-19.
  20. Friedman SL, Molecular mechanisms of hepatic fibrosis and principles of therapy. J Gastroenterol 1997;32(3):424-30. https://doi.org/10.1007/BF02934504
  21. 백용한. 간경변증: 간섬유화 기전 및 항섬유화 치료. 대한간학회지 2008;14(2s):7-13.
  22. Breitkopf K, Haas S, Wiercinska E, Singer MV, Dooley S. Anti-TGF -beta strategies for the treatment of chronic liver disease. Alcohol Clin Exp Res 2005;29(11 supple):121S-131S.
  23. Tangkijvanich P, Yee HF Jr. Cirrhosis--can we reverse hepatic fibrosis? Eur J Surg Suppl 2002;587:100-12.
  24. 한광협, 김승업. 간섬유화. 대한간학회지 2010;16(2s):47-56.
  25. Friedman SL, Bansal MB. Reversal of hepatic fibrosis---fact or fantasy? Hepatology 2006;43(suppl):82-8.
  26. An JH, Kim J, Seong J. Rodex signaling by ionizing radiation in mouse liver. Ann N Y Acad Sci 2004;1030:86-94. https://doi.org/10.1196/annals.1329.011
  27. Muller A, Machnik F, Zimmermann T, Schubert H. Thioacetamide -induced cirrhosis-like liver lesions in rats unusefulness and reliability of this animal model. Exp Pathol 1988;34(4):229-36. https://doi.org/10.1016/S0232-1513(88)80155-5
  28. Jang KT, Lee MS, Jung IP, Kim MR, Jang JJ. Effect of pehtoxifylline on liver fibrosis and cell cycle related proteins in thioacetamide -induced rat cirrhosis. Korean J Hepatol 2001;7:281-91.
  29. Kim KH, Bae JH, Cha SW, Han SS, Park KH, Jeong TC. Role of metabolic activation by cytochrome P450 in thioacetamide-induced suppression of antibody response in male BALB/c mice. Toxicol Lett 2000;114(1-3):225-35. https://doi.org/10.1016/S0378-4274(00)00168-5
  30. Yu Q, Stamenkovic I. Cell surface-localized matrix metalloproteinase-9 protelytically activates TGF-beta and promotestumor invasion and angiogrnesis. Gene Dev 2000;14(2):163-76.
  31. Friedman SL. Molecular regulation of hepatic fibrosis, an integrated cellular response to tissue injury. J Biol Chem 2000;275(4):2247-50. https://doi.org/10.1074/jbc.275.4.2247
  32. Bataller R and Brenner DA. Liver fibrosis. J Clin Invest 2005;115(2):209-18.
  33. Lee KS. Mechanism of liver fibrosis. In: Kim BS, ed. Liver fibrosis. Kunja; 2000, p. 27-42.
  34. 권영오. 간섬유화 치료의 최신지견. 대한간학회지 2005;11(2s):33-9.
  35. Pinzani M, Rombouts K. Liver fibrosis: from the bench to clinical targets. Dig Liver Lis 2004;36:231-42. https://doi.org/10.1016/j.dld.2004.01.003
  36. 경희대학교 부속한방병원. 경희한방처방집. 서울: 트윈기획; 1997, p. 244.
  37. 박용진, 김영철, 이장훈, 우홍정. 인진청간탕가미방이 간세포의 증식능력에 미치는 영향. 대한한의학회지 1998;19(1):145-64.
  38. 홍상훈, 이장훈, 우홍정. 인진청간탕가미방이 간세포활성세포주기 및 apoptosis에 미치는 영향. 대한한의학회지 1998;19(2):337-72.
  39. 강경태. 인진청간탕가미방이 간조직의 섬유화 억제에 미치는 영향. 대한한의학회지 2002;23(2):39-56.
  40. 승현석, 김영철, 이장훈, 우홍정. 인진청간탕이 간보호 및 섬유화 억제에 미치는 영향. 대한한방내과학회지 2003;24(1):21-32.
  41. 김영철, 이장훈, 우홍정. 인진청간탕이 흰쥐의 간장 비실질세포의 procollagen 합성 억제에 미치는 효과에 관한 연구. 대한한방내과학회지 2003;24(4):817-25.
  42. 박상백, 김영철, 이장훈, 우홍정. 인진청간탕이 DMN 유발 간섬유화와 단백질 발현에 미치는 영향. 대한한방내과학회지 2008;29(1):200-18.
  43. 박신명, 김영철, 이장훈, 우홍정. TAA로 유발된 간섬유화 동물 모델에서 인진청간탕의 효과. 대한한방내과학회지 2009;30(2):270-87.
  44. 이홍일. 인진청간탕이 간성상세포의 섬유화억제에 미치는 영향. 대한한방내과학회지 2009;30(1):74-84.
  45. 승현석. YBR의 간섬유화 억제 효과에 관한 연구. 대한한방내과학회지 2010;31(2):126-42.
  46. Li X, Benjamin IS, Alexander B. Reproducible production of thioacetamide-induced macronodular cirrhosis in the rat with no mortality. J Hepatol 2002;36(4):488-93. https://doi.org/10.1016/S0168-8278(02)00011-9
  47. Perez Tamayo R. Is cirrhosis of the liver experimentally produced by CCl4 and adequate model of human cirrhosis? Hepatology 1988;3(1):112-20.