Journal of Gastroenterology

, Volume 41, Issue 8, pp 772–778 | Cite as

Eupatilin attenuates bile acid-induced hepatocyte apoptosis

  • Su Cheol Park
  • Jung-Hwan Yoon
  • Won Kim
  • Geum-Youn Gwak
  • Kang Mo Kim
  • Sung-Hee Lee
  • Soo-Mi Lee
  • Hyo-Suk Lee
Article
Received:
Accepted:

Abstract

Background

In cases of cholestasis, bile acids induce hepatocyte apoptosis by activating death receptor-mediated apoptotic signaling cascades. Eupatilin (5,7-dihydroxy-3,4,6-trimethoxyflavone) is a pharmacologically active ingredient found in Artemisia asiatica and exhibits cytoprotective effects against experimentally induced gastrointestinal, pancreatic, and hepatic damage. This study was undertaken to examine if eupatilin modulates bile acid-induced hepatocyte apoptosis.

Methods

Huh-BAT cells, a human hepatocellular carcinoma cell line stably transfected with a bile acid transporter, were used in this study. Apoptosis was quantified using 4′,6-diamidino-2-phenylindole dihydrochloride staining, and its signaling cascades were explored by immunoblot analysis. Kinase signaling was evaluated by immunoblotting and by using selective inhibitors. Eupatilin's in vivo effect on bile acid-induced hepatocyte apoptosis was explored in bile duct-ligated rats.

Results

Eupatilin significantly reduced bile acid-mediated hepatocyte apoptosis by attenuating bile acid-induced caspase 8 cleavage. Eupatilin diminished the bile acid-induced activation of mitogen-activated protein kinases, including p38 mitogen-activated protein kinase and c-Jun N-terminal kinase. In particular, the eupatilin-mediated inhibition of bile acid-induced c-Jun N-terminal kinase activation was found to be responsible for attenuating caspase 8 cleavage. Moreover, eupatilin diminished hepatocyte apoptosis in bile duct-ligated rats.

Conclusions

Eupatilin attenuates bile acid-induced hepatocyte apoptosis by suppressing bile acid-induced kinase activation. Therefore, eupatilin might be therapeutically efficacious in a variety of human liver diseases associated with cholestasis.

Key words

eupatilin hepatocyte bile acid apoptosis c-Jun N-terminal kinase 
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References

  1. 1.
    Trauner, M, Meier, PJ, Boyer, JL 1998Molecular pathogenesis of cholestasisN Engl J Med339121727PubMedCrossRefGoogle Scholar
  2. 2.
    Miyoshi, H, Rust, C, Roberts, PJ, Burgart, LJ, Gores, GJ 1999Hepatocyte apoptosis after bile duct ligation in the mouse involves FasGastroenterology11766977PubMedCrossRefGoogle Scholar
  3. 3.
    Higuchi, H, Gores, GJ 2003Bile acid regulation of hepatic physiology: IV. Bile acids and death receptorsAm J Physiol Gastrointest Liver Physiol284G7348PubMedGoogle Scholar
  4. 4.
    Sodeman, T, Bronk, SF, Roberts, PJ, Miyoshi, H, Gores, GJ 2000Bile salts mediate hepatocyte apoptosis by increasing cell surface trafficking of FasAm J Physiol Gastrointest Liver Physiol278G9929PubMedGoogle Scholar
  5. 5.
    Faubion, WA, Guicciardi, ME, Miyoshi, H, Bronk, SF, Roberts, PJ, Svingen, PA,  et al. 1999Toxic bile salts induce rodent hepatocyte apoptosis via direct activation of FasJ Clin Invest10313745PubMedCrossRefGoogle Scholar
  6. 6.
    Yoon, JH, Gores, GJ 2002Death receptor-mediated apoptosis and the liverJ Hepatol3740010PubMedCrossRefGoogle Scholar
  7. 7.
    Higuchi, H, Bronk, SF, Takikawa, Y, Werneburg, N, Takimoto, R, El-Deiry, W,  et al. 2001The bile acid glycochenodeoxycholate induces trail-receptor 2/DR5 expression and apoptosisJ Biol Chem276386108PubMedCrossRefGoogle Scholar
  8. 8.
    Higuchi, H, Miyoshi, H, Bronk, SF, Zhang, H, Dean, N, Gores, GJ 2001Bid antisense attenuates bile acid-induced apoptosis and cholestatic liver injuryJ Pharmacol Exp Ther29986673PubMedGoogle Scholar
  9. 9.
    Canbay, A, Feldstein, A, Baskin-Bey, E, Bronk, SF, Gores, GJ 2004The caspase inhibitor IDN-6556 attenuates hepatic injury and fibrosis in the bile duct ligated mouseJ Pharmacol Exp Ther30811916PubMedCrossRefGoogle Scholar
  10. 10.
    Seol, SY, Kim, MH, Ryu, JS, Choi, MG, Shin, DW, Ahn, BO 2004DA-9601 for erosive gastritis: results of a double-blind placebo-controlled phase III clinical trialWorld J Gastroenterol10237982PubMedGoogle Scholar
  11. 11.
    Huh, K, Kwon, TH, Shin, US, Kim, WB, Ahn, BO, Oh, TY,  et al. 2003Inhibitory effects of DA-9601 on ethanol-induced gastrohemorrhagic lesions and gastric xanthine oxidase activity in ratsJ Ethnopharmacol8826973PubMedCrossRefGoogle Scholar
  12. 12.
    Hahm, KB, Kim, JH, You, BM, Kim, YS, Cho, SW, Yim, H,  et al. 1998Induction of apoptosis with an extract of Artemisia asiatica attenuates the severity of cerulein-induced pancreatitis in ratsPancreas171537PubMedCrossRefGoogle Scholar
  13. 13.
    Ahn, BO, Ko, KH, Oh, TY, Cho, H, Kim, WB, Lee, KJ,  et al. 2001Efficacy of use of colonoscopy in dextran sulfate sodium induced ulcerative colitis in rats: the evaluation of the effects of antioxidant by colonoscopyInt J Colorectal Dis1617481PubMedCrossRefGoogle Scholar
  14. 14.
    Oh, TY, Lee, JS, Ahn, BO, Cho, H, Kim, WB, Kim, YB,  et al. 2001Oxidative stress is more important than acid in the pathogenesis of reflux oesophagitis in ratsGut4936471PubMedCrossRefGoogle Scholar
  15. 15.
    Lee, JS, Oh, TY, Ahn, BO, Cho, H, Kim, WB, Kim, YB,  et al. 2001Involvement of oxidative stress in experimentally induced reflux esophagitis and Barrett's esophagus: clue for the chemoprevention of esophageal carcinoma by antioxidantsMutat Res480–481189200PubMedGoogle Scholar
  16. 16.
    Cheong, JY, Oh, TY, Lee, KM, Kim do, H, Ahn, BO, Kim, WB,  et al. 2002Suppressive effects of antioxidant DA-9601 on hepatic fibrosis in rats (in Korean with English abstract)Taehan Kan Hakhoe Chi (Korean J Hepatol)843647Google Scholar
  17. 17.
    Ryu, BK, Ahn, BO, Oh, TY, Kim, SH, Kim, WB, Lee, EB 1998Studies on protective effect of DA-9601, Artemisia asiatica extract, on acetaminophen- and CCl4-induced liver damage in ratsArch Pharm Res2150813PubMedCrossRefGoogle Scholar
  18. 18.
    Dent, P, Fang, Y, Gupta, S, Studer, E, Mitchell, C, Spiegel, S,  et al. 2005Conjugated bile acids promote ERK1/2 and AKT activation via a pertussis toxin-sensitive mechanism in murine and human hepatocytesHepatology4212919PubMedCrossRefGoogle Scholar
  19. 19.
    Rodrigues, CM, Fan, G, Wong, PY, Kren, BT, Steer, CJ 1998Ursodeoxycholic acid may inhibit deoxycholic acid-induced apoptosis by modulating mitochondrial transmembrane potential and reactive oxygen species productionMol Med416578PubMedCrossRefGoogle Scholar
  20. 20.
    Yerushalmi, B, Dahl, R, Devereaux, MW, Gumpricht, E, Sokol, RJ 2001Bile acid-induced rat hepatocyte apoptosis is inhibited by antioxidants and blockers of the mitochondrial permeability transitionHepatology3361626PubMedCrossRefGoogle Scholar
  21. 21.
    Sokol, RJ, Straka, MS, Dahl, R, Devereaux, MW, Yerushalmi, B, Gumpricht, E,  et al. 2001Role of oxidant stress in the permeability transition induced in rat hepatic mitochondria by hydrophobic bile acidsPediatr Res4951931PubMedGoogle Scholar
  22. 22.
    Xia, Z, Dickens, M, Raingeaud, J, Davis, RJ, Greenberg, ME 1995Opposing effects of ERK and JNK-p38 MAP kinases on apoptosisScience270132631PubMedGoogle Scholar
  23. 23.
    Grambihler, A, Higuchi, H, Bronk, SF, Gores, GJ 2003cFLIP-L inhibits p38 MAPK activation: an additional anti-apoptotic mechanism in bile acid-mediated apoptosisJ Biol Chem278268317PubMedCrossRefGoogle Scholar
  24. 24.
    Reinehr, R, Graf, D, Haussinger, D 2003Bile salt-induced hepatocyte apoptosis involves epidermal growth factor receptor-dependent CD95 tyrosine phosphorylationGastroenterology12583953PubMedCrossRefGoogle Scholar
  25. 25.
    Schoemaker, MH, Conde de la Rosa, L, Buist-Homan, M, Vrenken, TE, Havinga, R, Poelstra, K,  et al. 2004Tauroursodeoxycholic acid protects rat hepatocytes from bile acid-induced apoptosis via activation of survival pathwaysHepatology39156373PubMedCrossRefGoogle Scholar
  26. 26.
    Kummer, JL, Rao, PK, Heidenreich, KA 1997Apoptosis induced by withdrawal of trophic factors is mediated by p38 mitogen-activated protein kinaseJ Biol Chem272204904PubMedCrossRefGoogle Scholar
  27. 27.
    Reinehr, R, Becker, S, Wettstein, M, Haussinger, D 2004Involvement of the Src family kinase yes in bile salt-induced apoptosisGastroenterology127154057PubMedCrossRefGoogle Scholar
  28. 28.
    Higuchi, H, Grambihler, A, Canbay, A, Bronk, SF, Gores, GJ 2004Bile acids up-regulate death receptor 5/TRAIL-receptor 2 expression via a c-Jun N-terminal kinase-dependent pathway involving Sp1J Biol Chem2795160PubMedCrossRefGoogle Scholar
  29. 29.
    Czaja, MJ 2003The future of GI and liver research: editorial perspectives. III. JNK/AP-1 regulation of hepatocyte deathAm J Physiol Gastrointest Liver Physiol284G8759PubMedGoogle Scholar
  30. 30.
    Higuchi, H, Yoon, JH, Grambihler, A, Werneburg, N, Bronk, SF, Gores, GJ 2003Bile acids stimulate cFLIP phosphorylation enhancing TRAIL-mediated apoptosisJ Biol Chem27845461PubMedCrossRefGoogle Scholar
  31. 31.
    Wagner, M, Fickert, P, Zollner, G, Fuchsbichler, A, Silbert, D, Tsybrovskyy, O,  et al. 2003Role of farnesoid X receptor in determining hepatic ABC transporter expression and liver injury in bile duct-ligated miceGastroenterology12582538PubMedCrossRefGoogle Scholar
  32. 32.
    Guicciardi, ME, Gores, GJ 2005Cholestatic hepatocellular injury: what do we know and how should we proceedJ Hepatol42297300PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Tokyo 2006

Authors and Affiliations

  • Su Cheol Park
    • 1
  • Jung-Hwan Yoon
    • 1
  • Won Kim
    • 1
  • Geum-Youn Gwak
    • 1
  • Kang Mo Kim
    • 2
  • Sung-Hee Lee
    • 1
  • Soo-Mi Lee
    • 1
  • Hyo-Suk Lee
    • 1
  1. 1.Department of Internal Medicine and Liver Research InstituteSeoul National University College of MedicineSeoulRepublic of Korea
  2. 2.Department of Internal MedicineAsan Medical Center, University of Ulsan College of MedicineUlsanRepublic of Korea

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