Journal of Natural Medicines

, Volume 66, Issue 1, pp 149–157 | Cite as

Davallialactone protects against adriamycin-induced cardiotoxicity in vitro and in vivo

  • Sankarganesh Arunachalam
  • Sun Young Kim
  • Sun Hwa Lee
  • Young Hee Lee
  • Min Sun Kim
  • Bong Sik Yun
  • Ho Keun Yi
  • Pyoung Han Hwang
Original Paper


Adriamycin (ADR) is a potent anticancer drug. Its clinical applications are limited due to its cardiotoxicity. Oxidative stress is responsible for cardiomyopathy induced by ADR. Previous studies have demonstrated that davallialactone (DAVA), extracted from mushroom Inonotus xeranticus, has potential antiplatelet aggregation activity and free radical scavenging properties. In this study, we investigated whether DAVA has protective effects against ADR-induced free radical accumulation and apoptosis in cardiac muscle cells and compared the effects of DAVA with N-acetylcysteine, a potent antioxidant. We evaluated the effect of DAVA on ADR-induced cytotoxicity by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and crystal violet staining, the reactive oxygen species (ROS) production by flow cytometry, and the expression of stress-related proteins like Cu/Zn superoxide dismutase (SOD), Mn-SOD, and the involvement of mitogen-activated protein kinase pathway by Western blot analysis. Apoptosis was assessed by nuclear condensation and the expression levels of pro-apoptotic proteins, such as caspase-3 and polyadenosine diphosphate-ribose polymerase (PARP). The cardio-protective effects of DAVA were also evaluated in an in vivo study in an animal model of ADR-induced acute cardiomyopathy. Our results showed that DAVA significantly increased the viability of doxorubicin-injured H9c2 cells and inhibited ADR-induced ROS production, apoptosis, and the expression of Cu/Zn SOD and Mn-SOD. DAVA also inhibited the expression of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), which was activated by ADR. In the in vivo animal model, treatment involving DAVA significantly reduced cardiomyocyte lesions. These results suggest that DAVA is a potentially protective agent for ADR-induced cardiotoxicity in cardiomyocytes and can be a potential candidate to protect against cardiotoxicity in ADR-treated cancer patients.


Davallialactone Adriamycin Cardiotoxicity Antioxidant effect 



The authors thank Professor S. Kamala-Kannan for the careful review of the manuscript and revision suggestions. This study was supported by a grant from the Korea Forest Service (S121010L090120) and in part by a grant of the Clinical Trial Center for Functional Foods of Chonbuk National University Hospital.


  1. 1.
    Deng S, Kruger A, Kleschyov AL, Kalinowski L, Daiber A, Wojnowski L (2007) Gp91phox-containing NAD(P)H oxidase increases superoxide formation by doxorubicin and NADPH. Free Radic Biol Med 42:466–473PubMedCrossRefGoogle Scholar
  2. 2.
    Spallarossa P, Altieri P, Garibaldi S, Ghigliotti G, Barisione C, Manca V, Fabbi P, Ballestrero A, Brunelli C, Barsotti A (2006) Matrix metalloproteinase-2 and -9 are induced differently by doxorubicin in H9c2 cells: the role of MAP kinases and NAD(P)H oxidase. Cardiovasc Res 69:736–745PubMedCrossRefGoogle Scholar
  3. 3.
    Kalyanaraman B, Joseph J, Kalivendi S, Wang S, Konorov EA, Kotamraju S (2002) Doxorubicin-induced apoptosis: implications in cardiotoxicity. Mol Cell Biochem 234–235:119–124PubMedCrossRefGoogle Scholar
  4. 4.
    Kotamraju S, Konorev EA, Joseph J, Kalyanaraman B (2000) Doxorubicin-induced apoptosis in endothelial cells and cardiomyocytes is ameliorated by nitrone spin traps and ebselen. Role of reactive oxygen and nitrogen species. J Biol Chem 275:33585–33592PubMedCrossRefGoogle Scholar
  5. 5.
    Sawyer DB, Fukazawa R, Arstall MA, Kelly RA (1999) Daunorubicin-induced apoptosis in rat cardiac myocytes is inhibited by dexrazoxane. Circ Res 84:257–265PubMedGoogle Scholar
  6. 6.
    Haunstetter A, Izumo S (1998) Apoptosis: basic mechanisms and implications for cardiovascular disease. Circ Res 82:1111–1129PubMedGoogle Scholar
  7. 7.
    Young RC, Ozols RF, Myers CE (1981) The anthracycline antineoplastic drugs. N Engl J Med 305:139–153PubMedCrossRefGoogle Scholar
  8. 8.
    Lindequist U, Niedermeyer TH, Jülich WD (2005) The pharmacological potential of mushrooms. Evid Based Complement Alternat Med 2:285–299PubMedCrossRefGoogle Scholar
  9. 9.
    Ooi VE, Liu F (2000) Immunomodulation and anti-cancer activity of polysaccharide-protein complexes. Curr Med Chem 7:715–729PubMedGoogle Scholar
  10. 10.
    Imazeki R, Hongo T (1989) Colored illustrations of mushrooms of Japan. Hoikusha, OsakaGoogle Scholar
  11. 11.
    Kim SD, Lee IK, Lee WM, Cho JY, Park HJ, Oh JW, Park SC, Kim SK, Kwak YS, Yun BS, Rhee MH (2008) The mechanism of anti-platelet activity of davallialactone: involvement of intracellular calcium ions, extracellular signal-regulated kinase 2 and p38 mitogen-activated protein kinase. Eur J Pharmacol 584:361–367PubMedCrossRefGoogle Scholar
  12. 12.
    Lee YG, Lee WM, Kim JY, Lee JY, Lee IK, Yun BS, Rhee MH, Cho JY (2008) Src kinase-targeted anti-inflammatory activity of davallialactone from Inonotus xeranticus in lipopolysaccharide-activated RAW264.7 cells. Br J Pharmacol 154:852–863PubMedCrossRefGoogle Scholar
  13. 13.
    Lee IK, Yun BS (2006) Hispidin analogs from the mushroom Inonotus xeranticus and their free radical scavenging activity. Bioorg Med Chem Lett 16:2376–2379PubMedCrossRefGoogle Scholar
  14. 14.
    Tokarska-Schlattner M, Zaugg M, da Silva R, Lucchinetti E, Schaub MC, Wallimann T, Schlattner U (2005) Acute toxicity of doxorubicin on isolated perfused heart: response of kinases regulating energy supply. Am J Physiol Heart Circ Physiol 289:H37–H47PubMedCrossRefGoogle Scholar
  15. 15.
    DeAtley SM, Aksenov MY, Aksenova MV, Jordan B, Carney JM, Butterfield DA (1999) Adriamycin-induced changes of creatine kinase activity in vivo and in cardiomyocyte culture. Toxicology 134:51–62PubMedCrossRefGoogle Scholar
  16. 16.
    Kumar D, Kirshenbaum LA, Li T, Danelisen I, Singal PK (2001) Apoptosis in adriamycin cardiomyopathy and its modulation by probucol. Antioxid Redox Signal 3:135–145PubMedCrossRefGoogle Scholar
  17. 17.
    Nakamura T, Ueda Y, Juan Y, Katsuda S, Takahashi H, Koh E (2000) Fas-mediated apoptosis in adriamycin-induced cardiomyopathy in rats: in vivo study. Circulation 102:572–578PubMedGoogle Scholar
  18. 18.
    Sugden PH, Clerk A (1998) Regulation of mitogen-activated protein kinase cascades in the heart. Adv Enzyme Regul 38:87–98PubMedCrossRefGoogle Scholar
  19. 19.
    Xia Z, Dickens M, Raingeaud J, Davis RJ, Greenberg ME (1995) Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science 270:1326–1331PubMedCrossRefGoogle Scholar
  20. 20.
    Wang X, Martindale JL, Liu Y, Holbrook NJ (1998) The cellular response to oxidative stress: influences of mitogen-activated protein kinase signalling pathways on cell survival. Biochem J 333:291–300PubMedGoogle Scholar
  21. 21.
    Mizukami Y, Okamura T, Miura T, Kimura M, Mogami K, Todoroki-Ikeda N, Kobayashi S, Matsuzaki M (2001) Phosphorylation of proteins and apoptosis induced by c-Jun N-terminal kinase1 activation in rat cardiomyocytes by H2O2 stimulation. Biochim Biophys Acta 1540:213–220PubMedCrossRefGoogle Scholar
  22. 22.
    Torti FM, Bristow MR, Howes AE, Aston D, Stockdale FE, Carter SK, Kohler M, Brown BW Jr, Billingham ME (1983) Reduced cardiotoxicity of doxorubicin delivered on a weekly schedule. Assessment by endomyocardial biopsy. Ann Intern Med 99:745–749PubMedGoogle Scholar
  23. 23.
    Lores Arnaiz S, Llesuy S (1993) Oxidative stress in mouse heart by antitumoral drugs: a comparative study of doxorubicin and mitoxantrone. Toxicology 77:31–38PubMedCrossRefGoogle Scholar
  24. 24.
    Van Vleet JF, Ferrans VJ, Weirich WE (1980) Cardiac disease induced by chronic adriamycin administration in dogs and an evaluation of vitamin E and selenium as cardioprotectants. Am J Pathol 99:13–42PubMedGoogle Scholar
  25. 25.
    Liu X, Chen Z, Chua CC, Ma YS, Youngberg GA, Hamdy R, Chua BH (2002) Melatonin as an effective protector against doxorubicin-induced cardiotoxicity. Am J Physiol Heart Circ Physiol 283:H254–H263PubMedGoogle Scholar
  26. 26.
    Yamanaka S, Tatsumi T, Shiraishi J, Mano A, Keira N, Matoba S, Asayama J, Fushiki S, Fliss H, Nakagawa M (2003) Amlodipine inhibits doxorubicin-induced apoptosis in neonatal rat cardiac myocytes. J Am Coll Cardiol 41:870–878PubMedCrossRefGoogle Scholar
  27. 27.
    Andrieu-Abadie N (2004) Cardiotoxicity of anthracyclines: mechanisms and pharmacologic targets for prevention. Therapie 59(1):121–126PubMedCrossRefGoogle Scholar
  28. 28.
    Yen HC, Oberley TD, Gairola CG, Szweda LI, St Clair DK (1999) Manganese superoxide dismutase protects mitochondrial complex I against adriamycin-induced cardiomyopathy in transgenic mice. Arch Biochem Biophys 362:59–66PubMedCrossRefGoogle Scholar
  29. 29.
    Daosukho C, Chen Y, Noel T, Sompol P, Nithipongvanitch R, Velez JM, Oberley TD, St Clair DK (2007) Phenylbutyrate, a histone deacetylase inhibitor, protects against Adriamycin-induced cardiac injury. Free Radic Biol Med 42:1818–1825PubMedCrossRefGoogle Scholar
  30. 30.
    Kumar D, Lou H, Singal PK (2002) Oxidative stress and apoptosis in heart dysfunction. Herz 27:662–668PubMedCrossRefGoogle Scholar

Copyright information

© The Japanese Society of Pharmacognosy and Springer 2011

Authors and Affiliations

  • Sankarganesh Arunachalam
    • 1
    • 2
  • Sun Young Kim
    • 1
    • 2
  • Sun Hwa Lee
    • 3
  • Young Hee Lee
    • 4
  • Min Sun Kim
    • 1
  • Bong Sik Yun
    • 5
  • Ho Keun Yi
    • 4
  • Pyoung Han Hwang
    • 1
    • 2
  1. 1.Department of PediatricsChonbuk National University HospitalJeonjuRepublic of Korea
  2. 2.Research Institute of Clinical MedicineChonbuk National UniversityJeonjuRepublic of Korea
  3. 3.Division of Cardiology, Department of Internal Medicine, School of MedicineChonbuk National UniversityJeonjuRepublic of Korea
  4. 4.Department of Biochemistry, School of DentistryChonbuk National UniversityJeonjuRepublic of Korea
  5. 5.Division of Biotechnology, College of Environmental and Bioresources SciencesChonbuk National UniversityJeonjuRepublic of Korea

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