Investigational New Drugs

, Volume 29, Issue 2, pp 380–391

Chemopreventive doses of resveratrol do not produce cardiotoxicity in a rodent model of hepatocellular carcinoma

  • Daniel J. Luther
  • Vahagn Ohanyan
  • Patricia E. Shamhart
  • Cheryl M. Hodnichak
  • Hamayak Sisakian
  • Tristan D. Booth
  • J. Gary Meszaros
  • Anupam Bishayee


Hepatocellular carcinoma (HCC), one of the most lethal cancers, results in more than one million fatalities worldwide every year. In view of the limited therapeutic alternatives and poor prognosis of liver cancer, preventive control approaches, notably chemoprevention, have been considered to be the best strategy in lowering the present prevalence of the disease. Resveratrol, a naturally occurring antioxidant and antiinflammatory agent found in grapes and red wine, inhibits carcinogenesis with a pleiotropic mode of action. Recently, we have reported that dietary resveratrol significantly prevents chemically-induced liver tumorigenesis in rats. One of the mechanisms of resveratrol-mediated chemoprevention of hepatocarcinogenesis could be related to its antiinflammatory action through hepatic cyclooxygenase (COX-2) inhibition. Although several COX-2 inhibitors are known to exert chemopreventive efficacy, not all are considered ideal candidates for chemoprevention due to the risk of adverse cardiovascular events. Accordingly, the objective of the present study was to evaluate the role of resveratrol on cardiac performance during experimental hepatocarcinogenesis initiated with diethylnitrosamine and promoted by phenobarbital. Rats had free access to diet supplemented with resveratrol four weeks before the carcinogen injection and 14 weeks thereafter. The cardiotoxicity of resveratrol was assessed by monitoring the cardiac function using transthoracic echocardiography as well as Western blot analysis of cardiac tissue. Long-term dietary administration of resveratrol dose-dependently suppressed hepatic tumor multiplicity, the principal endpoint for evaluating the chemopreventive potential of a candidate agent. The chemopreventive effects of resveratrol were also reflected in histopathological assessment of hepatic tissues. Resveratrol did not exhibit any cardiotoxicity but rather improved the cardiac function in a dose-responsive fashion. Our results indicate that resveratrol-mediated chemoprevention of rat liver carcinogenesis is devoid of any adverse cardiovascular events. Resveratrol may be developed as a chemopreventive as well as therapeutic drug for human HCC.


Cardiac function Chemoprevention Diethylnitrosamine Echocardiography Hepatocarcinogenesis Resveratrol 


  1. 1.
    Feitelson MA, Sun B, Tufan NLS, Liu J, Pan J, Lian Z (2002) Genetic mechanisms of hepatocarcinogenesis. Oncogene 21:2593–2604CrossRefPubMedGoogle Scholar
  2. 2.
    Llovet JM, Burroughs A, Buix J (2003) Hepatocellular carcinoma. Lancet 362:1907–1917CrossRefPubMedGoogle Scholar
  3. 3.
    El-Serag HB (2004) Hepatocellular carcinoma: recent trends in the United States. Gastroenterology 127:S27–S34CrossRefPubMedGoogle Scholar
  4. 4.
    American Cancer Society (2009) Cancer facts and figures 2009. American Cancer Society, AtlantaGoogle Scholar
  5. 5.
    Bartsch H, Montesano R (1984) Relevance of nitrosamines to human cancer. Carcinogenesis 5:1381–1393CrossRefPubMedGoogle Scholar
  6. 6.
    Kensler TW, Egner PA, Wang JB, Zhu YR, Zhang BC, Lu PX et al (2004) Chemoprevention of hepatocellular carcinoma in aflatoxin endemic areas. Gastroenterology 127:S310–S318CrossRefPubMedGoogle Scholar
  7. 7.
    Pang R, Tse E, Poon TP (2006) Molecular pathways in hepatocellular carcinoma. Cancer Lett 240:157–169CrossRefPubMedGoogle Scholar
  8. 8.
    Schütte K, Bornscein J, Malfertheiner P (2009) Hepatocellular carcinoma—epidemiological trends and risk factors. Dig Dis 27:80–92PubMedGoogle Scholar
  9. 9.
    Ribes J, Clèries R, Esteban L, Moreno V, Bosch FX (2008) The influence of alcohol consumption and hepatitis B and C infections on the risk of liver cancer in Europe. J Hepatol 49:233–242CrossRefPubMedGoogle Scholar
  10. 10.
    Kensler TW, Quian GS, Chen JG, Groopman JD (2003) Translational strategies for cancer prevention in liver. J Natl Cancer Inst 3:321–329Google Scholar
  11. 11.
    Okuno M, Kojima S, Moriwaki H (2001) Chemoprevention of hepatocellular carcinoma: concept, progress and perspectives. J Gastroenterol Hepatol 16:1329–1335CrossRefPubMedGoogle Scholar
  12. 12.
    Yates MS, Kensler TW (2007) Keap1 eye on the target: chemoprevention of liver cancer. Acta Pharmacol Sin 28:1331–1342CrossRefPubMedGoogle Scholar
  13. 13.
    Lippman SM, Hong WK (2002) Cancer chemoprevention science and practice. Cancer Res 62:5119–5125PubMedGoogle Scholar
  14. 14.
    World Cancer Research Fund/American Institute for Cancer Research (2007) Food, nutrition, physical activity, and the prevention of cancer: a global perspective. AICR, WashingtonGoogle Scholar
  15. 15.
    Naithani R, Huma LC, Moriarty RM, McCormick DL, Mehta RG (2008) Comprehensive review of cancer chemopreventive agents evaluated in experimental carcinogenesis models and clinical trials. Curr Med Chem 15:1044–1071CrossRefPubMedGoogle Scholar
  16. 16.
    Mann CD, Neal CP, Garcea G, Manson MM, Dennison AR, Berry DP (2009) Phytochemicals as potential chemopreventive and chemotherapeutic agents in hepatocarcinogenesis. Eur J Cancer Prev 18:13–25CrossRefPubMedGoogle Scholar
  17. 17.
    Harikumar KB, Aggarwal BB (2008) Resveratrol, a multitargeted agent for age associated chronic diseases. Cell cycle 7:1020–1037CrossRefPubMedGoogle Scholar
  18. 18.
    Fremont L (2000) Biological effects of resveratrol. Life Sci 66:663–673CrossRefPubMedGoogle Scholar
  19. 19.
    Kopp P (1998) Resveratrol, a phytoestrogen found in red wine. A possible explanation for the conundrum of the ‘French paradox’? Eur J Endocrinol 138:619–620CrossRefPubMedGoogle Scholar
  20. 20.
    Baur JA, Sinclair DA (2006) Therapeutic potential of resveratrol: the in vivo evidence. Nat Rev Drug Discov 5:493–506CrossRefPubMedGoogle Scholar
  21. 21.
    Shankar S, Singh G, Srivastava RK (2007) Chemoprevention by resveratrol: molecular mechanisms and therapeutic potential. Front Biosci 12:4839–4854CrossRefPubMedGoogle Scholar
  22. 22.
    Saiko P, Szakmary A, Jaeger W, Szekeres T (2008) Resveratrol and its analogs: defense against cancer, coronary disease and neurodegenerative maladies or just a fad? Mutat Res 658:68–94CrossRefPubMedGoogle Scholar
  23. 23.
    Shakibaei M, Harikumar KB, Aggarwal BB (2009) Resveratrol addiction: to die or not to die. Mol Nutr Food Res 53:115–128CrossRefPubMedGoogle Scholar
  24. 24.
    Athar M, Back JH, Kopelovich L, Bickers DR, Kim AL (2009) Multiple molecular targets of resveratrol: anti-carcinogenic mechanisms. Arch Biochem Biophys 486:95–102CrossRefPubMedGoogle Scholar
  25. 25.
    Pirola L, Fröjdö S (2008) Resveratrol: one molecule, many targets. IUBMB Life 60:323–332CrossRefPubMedGoogle Scholar
  26. 26.
    Aggarwal BB, Bhardwaj A, Aggarwal RS, Seeram NP, Shishodia S, Takada Y (2004) Role of resveratrol in prevention and therapy of cancer: preclinical and clinical studies. Anticancer Res 24:2783–2840PubMedGoogle Scholar
  27. 27.
    Kundu JK, Surh Y-J (2008) Cancer chemopreventive and therapeutic potential of resveratrol: mechanistic perspectives. Cancer Lett 269:243–261CrossRefPubMedGoogle Scholar
  28. 28.
    Bishayee A (2009) Cancer prevention and treatment with resveratrol: from rodent studies to clinical trials. Cancer Prev Res 2:409–418CrossRefGoogle Scholar
  29. 29.
    Bishayee A, Dhir N (2009) Resveratrol-mediated chemoprevention of diethylnitrosamine-initiated hepatocarcinogenesis: inhibition of cell proliferation and induction of apoptosis. Chem-Biol Interact 179:131–144CrossRefPubMedGoogle Scholar
  30. 30.
    Penumathsa SV, Maulik N (2009) Resveratrol: a promising agent in promoting cardioprotection against coronary heart disease. Can J Physiol Pharmacol 87:275–286CrossRefPubMedGoogle Scholar
  31. 31.
    Ray PS, Maulik G, Cordis GA, Bertelli AA, Bertelli A, Das DK (1999) The red wine antioxidant resveratrol protects isolated rat hearts from ischemia reperfusion injury. Free Radic Biol Med 27:160–169CrossRefPubMedGoogle Scholar
  32. 32.
    Bradamante S, Piccinini F, Barenghi L, Bertelli AA, De Jonge R, Beemster P et al (2000) Does resveratrol induce pharmacological preconditioning? Int J Tissue React 22:1–4PubMedGoogle Scholar
  33. 33.
    Hattori R, Otani H, Maulik N, Das DK (2002) Pharmacological preconditioning with resveratrol: role of nitric oxide. Am J Physiol Heart Circ Physiol 282:H1988–H1995PubMedGoogle Scholar
  34. 34.
    Hung LM, Su MJ, Chen JK (2004) Resveratrol protects myocardial ischemia-reperfusion injury through both NO-dependent and NO-independent mechanisms. Free Radic Biol Med 36:774–781CrossRefPubMedGoogle Scholar
  35. 35.
    Chen CK, Pace-Asciak CR (1996) Vasorelaxing activity of resveratrol and quercetin in isolated rat aorta. Gen Pharmacol 27:363–366PubMedGoogle Scholar
  36. 36.
    Bertelli AA, Giovannini L, Giannessi D, Migliori M, Bernini W, Fregoni M et al (1995) Antiplatelet activity of synthetic and natural resveratrol in red wine. Int J Tissue React 17:1–3PubMedGoogle Scholar
  37. 37.
    Das S, Das DK (2007) Anti-inflammatory responses of resveratrol. Inflamm Allergy Drug Targets 6:168–173CrossRefPubMedGoogle Scholar
  38. 38.
    Olson ER, Naugle JE, Zhang X, Bomser JA, Meszaros JG (2005) Inhibition of cardiac fibroblast proliferation and myofibroblast differentiation by resveratrol. Am J Physiol Heart Circ Physiol 288:H1131–H1138CrossRefPubMedGoogle Scholar
  39. 39.
    Das S, Fraga CG, Das DK (2006) Cardioprotective effect of resveratrol via HO-1 expression involves p38 map kinase and PI-3-kinase signaling, but does not involve NFkappaB. Free Radic Res 40:1066–1075CrossRefPubMedGoogle Scholar
  40. 40.
    Dudley J, Das S, Mukherjee S, Das DK (2009) Resveratrol, a unique phytoalexin present in red wine, delivers either survival signal or death signal to the ischemic myocardium depending on dose. J Nutr Biochem 20:443–452CrossRefPubMedGoogle Scholar
  41. 41.
    Wu T (2006) Cyclooxygenase-2 in hepatocellular carcinoma. Cancer Treat Rev 32:28–44CrossRefPubMedGoogle Scholar
  42. 42.
    Yildirim Y, Ozyilkan O, Bilezikci B, Akcali Z, Haberal M (2008) Lack of influence of cyclooxygenase-expression in hepatocellular carcinomas on patient survival. Asian Pacific J Cancer Prev 9:295–298Google Scholar
  43. 43.
    Giannitrapani L, Ingrao S, Soresi M, Maria Florena A, La Spada E, Sandonato L et al (2009) Cyclooxygenase-2 expression in chronic liver diseases and hepatocellular carcinoma. Ann NY Acad Sci 1155:293–299CrossRefPubMedGoogle Scholar
  44. 44.
    Hu K-Q (2002) Rationale and feasibility of chemoprevention of hepatocellular carcinoma by cyclooxygenase-2 inhibitors. J Lab Clin Med 139:234–243CrossRefPubMedGoogle Scholar
  45. 45.
    Koga H (2003) Hepatocelluar carcinoma: is there a potential for chemoprevention using cyclooxygenase-2 inhibitors. Cancer 98:661–667CrossRefPubMedGoogle Scholar
  46. 46.
    Bishayee A, Waghray A, Lotey R, Barnes KF, Darvesh AS, Bhatia D, Carroll RT (2009) Mechanisms of resveratrol-mediated chemoprevention of hepatocellular carcinogenesis: suppression of oxidative stress and inflammation. Eighth Annual International Conference on Frontiers in Cancer Prevention Research, December 6–9, 2009, Houston, TX, USAGoogle Scholar
  47. 47.
    Cervello M, Montalto G (2006) Cyclooxygenase in hepatocellular carcinoma. World J Gastroenterol 12:5113–5121PubMedGoogle Scholar
  48. 48.
    Fujimura T, Ohta T, Oyama K, Miyashita T, Miwa K (2007) Cycloxygenase-2 (COX-2) in carcinogenesis and selective COX-2 inhibitors for chemoprevention in gastrointestinal cancers. J Gastrointest Cancer 38:78–82CrossRefPubMedGoogle Scholar
  49. 49.
    Yona D, Arber N (2004) Coxibs and cancer prevention. J Cardiovasc Pharmacol 47:S76–S81CrossRefGoogle Scholar
  50. 50.
    Niles RM, Cook CP, Meadows GG, Fu Y-M, McLaughlin JL, Rankin GO (2006) Resveratrol is rapidly metabolized in athymic (Nu/Nu) mice and does not inhibit human melanoma xenograft tumor growth. J Nutr 136:2542–2546PubMedGoogle Scholar
  51. 51.
    Bushan KM, Rao GV, Soujanya T, Rao VJ, Saha S, Samanta A (2001) Photochemical E (trans)–>Z (cis) isomerization in substituted 1-naphthylacrylates. J Org Chem 66:681–688CrossRefPubMedGoogle Scholar
  52. 52.
    Bishayee A, Roy S, Chatterjee M (1999) Characterization of selective induction and alteration of xenobiotic biotransforming enzymes by vanadium during diethylnitrosamine-induced chemical rat liver carcinogenesis. Oncol Res 11:41–53PubMedGoogle Scholar
  53. 53.
    Bishayee A, Sarkar A, Chatterjee M (2000) Further evidence for chemopreventive potential of β-carotene against experimental hepatocarcinogenesis: diethylnitrosamine-initiated and phenobarbital promoted hepatocarcinogenesis is prevented more effectively by β-carotene than by retinoic acid. Nutr Cancer 37:89–98CrossRefPubMedGoogle Scholar
  54. 54.
    Bishayee A, Chatterjee M (1995) Inhibition of altered liver cell foci and persistent nodule growth by vanadium during diethylnitrosamine-induced hepatocarcinogenesis in rats. Anticancer Res 15:455–462PubMedGoogle Scholar
  55. 55.
    Stewart HL, Williams GM, Keysser CH, Lombart LS, Montali RJ (1980) Histological typing of liver tumors of the rat. J Natl Cancer Inst 64:177–204Google Scholar
  56. 56.
    Sahn DJ, DeMaria A, Kisslo J, Weyman A (1978) Recommendations regarding quantitation in M-mode echocardiography: results of a survey of echocardiographic measurements. Circulation 58:1072–1083PubMedGoogle Scholar
  57. 57.
    Bishayee A, Chatterjee M (1995) Inhibitory effect of vanadium on rat liver carcinogenesis initiated with diethylnitrosamine and promoted by phenobarbital. Brit J Cancer 71:1214–1220CrossRefPubMedGoogle Scholar
  58. 58.
    Chakraborty T, Chatterjee A, Rana A, Dhachinamoorthi D, Kumar PA, Chatterjee M (2007) Carcinogen-induced early molecular events and its implication in the initiation of chemical hepatocarcinogenesis in rats: chemopreventive role of vanadium on this process. Biochim Biophys Acta 1772:48–59PubMedGoogle Scholar
  59. 59.
    Lee JS, Chu IS, Mikaelyan A, Calvisi DF, Heo J, Reddy JK, Thorgeirsson SS (2004) Application of comparative functional genomics to identify best-fit mouse models to study human cancer. Nat Genetics 36:1306–1311CrossRefGoogle Scholar
  60. 60.
    Farber E, Sarma DS (1987) Hepatocarcinogenesis: a dynamic cellular perspective. Lab Invest 56:4–22PubMedGoogle Scholar
  61. 61.
    Williams GM (1980) The pathogenesis of rat liver cancer caused by chemical carcinogens. Biochim Biophys Acta 605:167–189PubMedGoogle Scholar
  62. 62.
    Chen T, Hwang H, Rose ME, Nines RG, Stoner GD (2006) Chemopreventive properties of black raspberries in N-nitrosomethylbenzylamine-induced rat esophageal tumorigenesis: down-regulation of cyclooxygenase-2, inducible nitric oxide synthase, and c-Jun. Cancer Res 66:2853–2859CrossRefPubMedGoogle Scholar
  63. 63.
    Di Stefano G, Fiume L, Bolondi L, Lanza M, Pariali M, Chieco P (2005) Enhanced uptake of lactosaminated human albumin by rat hepatocarcinomas: implications for an improved chemotherapy of primary liver tumours. Liver Int 25:854–860CrossRefPubMedGoogle Scholar
  64. 64.
    Waitzberz DL, Goncalves EL, Faintuch J, Bevilacqua LR, Rocha CL, Cdogni AM (1989) Effects of diets with different protein levels on the growth of Walker 256 carcinosarcoma in rats. Brazil J Med Biol Res 22:447–455Google Scholar
  65. 65.
    Husting SD, Kari FW (1999) The anti-carcinogenic effects of dietary restriction: mechanisms and future directions. Mutat Res 443:235–249Google Scholar
  66. 66.
    Whitsett T, Carpenter M, Lamartiniere CA (2006) Resveratrol, but not EGCG, in the diet suppresses DMBA-induced mammary cancer in rats. J Carcinog 5:15CrossRefPubMedGoogle Scholar
  67. 67.
    Crowell JA, Korytko PJ, Morrissey RL, Booth TD, Levine BS (2004) Resveratrol-associated renal toxicity. Toxicol Sci 82:614–619CrossRefPubMedGoogle Scholar
  68. 68.
    Williams LD, Burdock GA, Edwards JA, Beck M, Bausch J (2009) Safety studies conducted on high-purity trans-resveratrol in experimental animals. Food Chem Toxicol 47:2170–2182CrossRefPubMedGoogle Scholar
  69. 69.
    Bombardier C, Laine L, Reicin A, Shapiro D, Burgos-Vargas R, Davis B et al (2000) Comparison of upper gastrointestinal toxicity of rofecoxib and naproxen in patients with rheumatoid arthritis. N Eng J Med 343:1520–1528CrossRefGoogle Scholar
  70. 70.
    Mamdani M, Juurlink DN, Lee DS, Rochon PA, Kopp A, Naglie G et al (2004) Cyclo-oxygenase-2 inhibitors versus non-selective non-steroidal anti-inflammatory drugs and congestive heart failure outcomes in elderly patients: a population-based cohort study. Lancet 363:1751–1756CrossRefPubMedGoogle Scholar
  71. 71.
    Solomon DH, Schneeweiss S, Glynn RJ, Kiyota Y, Lenin R, Mogun H (2004) Relationship between selective cyclooxygenase-2 inhibitors and acute myocardial infarction in older adults. Circulation 109:2068–2073CrossRefPubMedGoogle Scholar
  72. 72.
    Baron JA, Sandler RS, Bresalier RS, Quan H, Riddell R, Lanas A et al (2006) A randomized trial of rofecoxib for the chemoprevention of colorectal adenomas. Gastroenterology 131:1674–1682CrossRefPubMedGoogle Scholar
  73. 73.
    Bertagnolli MM, Eagle CJ, Zauber AG, Redston M, Solomon SD, Kim K et al (2006) Celecoxib for the prevention of sporadic colorectal adenomas. N Engl J Med 355:873–884CrossRefPubMedGoogle Scholar
  74. 74.
    Burstein B, Majuy A, Clement R, Gosselin H, Poulin F, Ethier N et al (2007) Effects of resveratrol treatment on cardiac remodeling following myocardial infarction. J Pharmacol Exp Ther 323:916–23CrossRefPubMedGoogle Scholar
  75. 75.
    Lin JF, Lin SM, Chih CL, Nien MW, Su HH, Hu BR, Huang SS, Tsai SK (2008) Resveratrol reduces infarct size and improves ventricular function after myocardial ischemia in rats. Life Sci 89:313–317CrossRefGoogle Scholar
  76. 76.
    Juric D, Wojciechowski P, Das DK, Netticadan T (2007) Prevention of concentric hypertrophy and diastolic impairment in aortic-banded rats treated with resveratrol. Am J Physiol Heart Circ Physiol 292:H2138–H2143CrossRefPubMedGoogle Scholar
  77. 77.
    Chan AY, Dolinsky VW, Soltys CL, Viollet B, Baksh S, Light PE et al (2008) Resveratrol inhibits cardiac hypertrophy via AMP-activated kinase and Akt. J Biol Chem 283:24194–24201CrossRefPubMedGoogle Scholar
  78. 78.
    Tatlidede E, Sehirli O, Velioğlu-Oğünc A, Cetinel S, Yeğen BC, Yarat A et al (2009) Resveratrol treatment protects against doxorubicin-induced cardiotoxicity by alleviating oxidative damage. Free Radic Res 43:195–205CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Daniel J. Luther
    • 1
    • 2
  • Vahagn Ohanyan
    • 1
  • Patricia E. Shamhart
    • 1
    • 2
  • Cheryl M. Hodnichak
    • 1
  • Hamayak Sisakian
    • 3
  • Tristan D. Booth
    • 4
  • J. Gary Meszaros
    • 1
    • 2
  • Anupam Bishayee
    • 2
    • 5
  1. 1.Department of Integrative Medical SciencesNortheastern Ohio Universities Colleges of Medicine and PharmacyRootstownUSA
  2. 2.Graduate Program, School of Biomedical SciencesKent State UniversityKentUSA
  3. 3.Department of Internal Diseases 1, Cardiology Division, University Hospital 1Yerevan State Medical UniversityYerevanArmenia
  4. 4.Innovative Drug Development, Pharmascience Inc.MontréalCanada
  5. 5.Department of Pharmaceutical SciencesNortheastern Ohio Universities Colleges of Medicine and PharmacyRootstownUSA

Personalised recommendations