, Volume 12, Issue 11, pp 1953–1963

Effect of xanthohumol and isoxanthohumol on 3T3-L1 cell apoptosis and adipogenesis

  • Jeong-Yeh Yang
  • Mary Anne Della-Fera
  • Srujana Rayalam
  • Clifton A. Baile
Original Paper


Xanthohumol (XN), the chalcone from beer hops has several biological activities. XN has been shown to induce apoptosis in cancer cells and also has been reported to be involved in lipid metabolism. Based on these studies and our previous work with natural compounds, we hypothesized that XN and its isomeric flavanone, isoxanthohumol (IXN), would induce apoptosis in adipocytes through the mitochondrial pathway and would inhibit maturation of preadipocytes. Adipocytes were treated with various concentrations of XN or IXN. In mature adipocytes both XN and IXN decreased viability, increased apoptosis and increased ROS production, XN being more effective. Furthermore, the antioxidants ascorbic acid and 2-mercaptoethanol prevented XN and IXN-induced ROS generation and apoptosis. Immunoblotting analysis showed an increase in the levels of cytoplasmic cytochrome c and cleaved poly (ADP-ribose) polymerase (PARP) by XN and IXN. Concomitantly, we observed activation of the effectors caspase-3/7. In maturing preadipocytes both XN and IXN were effective in reducing lipid content, XN being more potent. Moreover, the major adipocyte marker proteins such as PPARγ, C/EBPα, and aP2 decreased after treatment with XN during the maturation period and that of DGAT1 decreased after treatment with XN and IXN. Taken together, our data indicate that both XN and IXN inhibit differentiation of preadipocytes, and induce apoptosis in mature adipocytes, but XN is more potent.


Reactive oxygen species Caspase-3/7 Mitochondrial membrane potential Peroxisome proliferator-activated receptor γ (PPARγ) CCAAT/enhancer binding protein α (C/EBPα) 


  1. 1.
    Kim HK, Nelson-Dooley C, Della-Fera MA et al (2006) Genistein decreases food intake, body weight, and fat pad weight and causes adipose tissue apoptosis in ovariectomized female mice. J Nutr 136:409–414PubMedGoogle Scholar
  2. 2.
    Lin J, Della-Fera MA, Baile CA (2005) Green tea polyphenol epigallocatechin gallate inhibits adipogenesis and induces apoptosis in 3T3-L1 adipocytes. Obes Res 13:982–990PubMedGoogle Scholar
  3. 3.
    Stevens JF, Taylor AW, Clawson JE, Deinzer ML (1999) Fate of xanthohumol and related prenylflavonoids from hops to beer. J Agric Food Chem 47:2421–2428PubMedCrossRefGoogle Scholar
  4. 4.
    Stevens JF, Taylor AW, Deinzer ML (1999) Quantitative analysis of xanthohumol and related prenylflavonoids in hops and beer by liquid chromatography-tandem mass spectrometry. J Chromatogr A 832:97–107PubMedCrossRefGoogle Scholar
  5. 5.
    Dietz BM, Kang YH, Liu G et al (2005) Xanthohumol isolated from Humulus lupulus Inhibits menadione-induced DNA damage through induction of quinone reductase. Chem Res Toxicol 18:1296–1305PubMedCrossRefGoogle Scholar
  6. 6.
    Gerhauser C, Alt A, Heiss E et al (2002) Cancer chemopreventive activity of Xanthohumol, a natural product derived from hop. Mol Cancer Ther 1:959–969PubMedGoogle Scholar
  7. 7.
    Goto K, Asai T, Hara S et al (2005) Enhanced antitumor activity of xanthohumol, a diacylglycerol acyltransferase inhibitor, under hypoxia. Cancer Lett 219:215–222PubMedCrossRefGoogle Scholar
  8. 8.
    Tabata N, Ito M, Tomoda H, Omura S (1997) Xanthohumols, diacylglycerol acyltransferase inhibitors, from Humulus lupulus. Phytochemistry 46:683–687PubMedCrossRefGoogle Scholar
  9. 9.
    Pan L, Becker H, Gerhauser C (2005) Xanthohumol induces apoptosis in cultured 40-16 human colon cancer cells by activation of the death receptor- and mitochondrial pathway. Mol Nutr Food Res 49:837–843PubMedCrossRefGoogle Scholar
  10. 10.
    Lust S, Vanhoecke B, Janssens A, Philippe J, Bracke M, Offner F (2005) Xanthohumol kills B-chronic lymphocytic leukemia cells by an apoptotic mechanism. Mol Nutr Food Res 49:844–850PubMedCrossRefGoogle Scholar
  11. 11.
    Casaschi A, Maiyoh GK, Rubio BK, Li RW, Adeli K, Theriault AG (2004) The chalcone xanthohumol inhibits triglyceride and apolipoprotein B secretion in HepG2 cells. J Nutr 134:1340–1346PubMedGoogle Scholar
  12. 12.
    Nozawa H (2005) Xanthohumol, the chalcone from beer hops (Humulus lupulus L.), is the ligand for farnesoid X receptor and ameliorates lipid and glucose metabolism in KK-A(y) mice. Biochem Biophys Res Commun 336:754–761PubMedCrossRefGoogle Scholar
  13. 13.
    Roncari DA, Lau DC, Kindler S (1981) Exaggerated replication in culture of adipocyte precursors from massively obese persons. Metabolism 30:425–427PubMedCrossRefGoogle Scholar
  14. 14.
    Dowell P, Flexner C, Kwiterovich PO, Lane MD (2000) Suppression of preadipocyte differentiation and promotion of adipocyte death by HIV protease inhibitors. J Biol Chem 275:41325–41332PubMedCrossRefGoogle Scholar
  15. 15.
    Prins JB, O’Rahilly S (1997) Regulation of adipose cell number in man. Clin Sci (Lond) 92:3–11Google Scholar
  16. 16.
    Prins JB, Walker NI, Winterford CM, Cameron DP (1994) Apoptosis of human adipocytes in vitro. Biochem Biophys Res Commun 201:500–507PubMedCrossRefGoogle Scholar
  17. 17.
    Ntambi JM, Young-Cheul K (2000) Adipocyte differentiation and gene expression. J Nutr 130:3122S–3126SPubMedGoogle Scholar
  18. 18.
    Gregoire FM, Smas CM, Sul HS (1998) Understanding adipocyte differentiation. Physiol Rev 78:783–809PubMedGoogle Scholar
  19. 19.
    Sun SY, Hail N Jr, Lotan R (2004) Apoptosis as a novel target for cancer chemoprevention. J Natl Cancer Inst 96:662–672PubMedCrossRefGoogle Scholar
  20. 20.
    Danial NN, Korsmeyer SJ (2004) Cell death: critical control points. Cell 116:205–219PubMedCrossRefGoogle Scholar
  21. 21.
    Reed JC (2004) Apoptosis mechanisms: implications for cancer drug discovery. Oncology (Williston Park) 18:11–20Google Scholar
  22. 22.
    Assuncao Guimaraes C, Linden R (2004) Programmed cell deaths. Apoptosis and alternative deathstyles. Eur J Biochem 271:1638–1650PubMedCrossRefGoogle Scholar
  23. 23.
    Hemati N, Ross SE, Erickson RL, Groblewski GE, MacDougald OA (1997) Signaling pathways through which insulin regulates CCAAT/enhancer binding protein alpha (C/EBPalpha) phosphorylation and gene expression in 3T3-L1 adipocytes. Correlation with GLUT4 gene expression. J Biol Chem 272:25913–25919PubMedCrossRefGoogle Scholar
  24. 24.
    Frankfurt OS (2004) Immunoassay for single-stranded DNA in apoptotic cells. Methods Mol Biol 282:85–101PubMedGoogle Scholar
  25. 25.
    Frankfurt OS, Krishan A (2001) Identification of apoptotic cells by formamide-induced dna denaturation in condensed chromatin. J Histochem Cytochem 49:369–378PubMedGoogle Scholar
  26. 26.
    Suryawan A, Hu CY (1993) Effect of serum on differentiation of porcine adipose stromal-vascular cells in primary culture. Comp Biochem Physiol Comp Physiol 105:485–492PubMedCrossRefGoogle Scholar
  27. 27.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  28. 28.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  29. 29.
    Jacobson MD, Raff MC (1995) Programmed cell death and Bcl-2 protection in very low oxygen. Nature 374:814–816PubMedCrossRefGoogle Scholar
  30. 30.
    Miranda CL, Stevens JF, Helmrich A et al (1999) Antiproliferative and cytotoxic effects of prenylated flavonoids from hops (Humulus lupulus) in human cancer cell lines. Food Chem Toxicol 37:271–285PubMedCrossRefGoogle Scholar
  31. 31.
    Colgate EC, Miranda CL, Stevens JF, Bray TM, Ho E (2007) Xanthohumol, a prenylflavonoid derived from hops induces apoptosis and inhibits NF-kappaB activation in prostate epithelial cells. Cancer Lett 246:201–209PubMedCrossRefGoogle Scholar
  32. 32.
    Buttke TM, Sandstrom PA (1994) Oxidative stress as a mediator of apoptosis. Immunol Today 15:7–10PubMedCrossRefGoogle Scholar
  33. 33.
    Yang JY, Della-Fera MA, Nelson-Dooley C, Baile CA (2006) Molecular mechanisms of apoptosis induced by ajoene in 3T3-L1 adipocytes. Obesity (Silver Spring) 14:388–397CrossRefGoogle Scholar
  34. 34.
    Ding WX, Shen HM, Ong CN (2000) Critical role of reactive oxygen species and mitochondrial permeability transition in microcystin-induced rapid apoptosis in rat hepatocytes. Hepatology 32:547–555PubMedCrossRefGoogle Scholar
  35. 35.
    Kroemer G, Zamzami N, Susin SA (1997) Mitochondrial control of apoptosis. Immunol Today 18:44–51PubMedCrossRefGoogle Scholar
  36. 36.
    Iijima T (2006) Mitochondrial membrane potential and ischemic neuronal death. Neurosci Res 55:234–243PubMedCrossRefGoogle Scholar
  37. 37.
    Chan WH, Wu CC, Yu JS (2003) Curcumin inhibits UV irradiation-induced oxidative stress and apoptotic biochemical changes in human epidermoid carcinoma A431 cells. J Cell Biochem 90:327–338PubMedCrossRefGoogle Scholar
  38. 38.
    Chan WH, Wu HJ (2004) Anti-apoptotic effects of curcumin on photosensitized human epidermal carcinoma A431 cells. J Cell Biochem 92:200–212PubMedCrossRefGoogle Scholar
  39. 39.
    Gorman A, McGowan A, Cotter TG (1997) Role of peroxide and superoxide anion during tumour cell apoptosis. FEBS Lett 404:27–33PubMedCrossRefGoogle Scholar
  40. 40.
    Koide T, Kamei H, Hashimoto Y, Kojima T, Hasegawa M (1996) Antitumor effect of hydrolyzed anthocyanin from grape rinds and red rice. Cancer Biother Radiopharm 11:273–277PubMedCrossRefGoogle Scholar
  41. 41.
    Lautier D, Lagueux J, Thibodeau J, Menard L, Poirier GG (1993) Molecular and biochemical features of poly (ADP-ribose) metabolism. Mol Cell Biochem 122:171–193PubMedCrossRefGoogle Scholar
  42. 42.
    Kawada T, Kamei Y, Fujita A et al (2000) Carotenoids and retinoids as suppressors on adipocyte differentiation via nuclear receptors. Biofactors 13:103–109PubMedGoogle Scholar
  43. 43.
    Tontonoz P, Hu E, Spiegelman BM (1994) Stimulation of adipogenesis in fibroblasts by PPAR gamma 2, a lipid-activated transcription factor. Cell 79:1147–1156PubMedCrossRefGoogle Scholar
  44. 44.
    Coleman RA, Lewin TM, Muoio DM (2000) Physiological and nutritional regulation of enzymes of triacylglycerol synthesis. Annu Rev Nutr 20:77–103PubMedCrossRefGoogle Scholar
  45. 45.
    Rizzo G, Disante M, Mencarelli A et al (2006) The farnesoid X receptor promotes adipocyte differentiation and regulates adipose cell function in vivo. Mol pharmacol 70:1164–1173PubMedCrossRefGoogle Scholar
  46. 46.
    Yang JY, Della-Fera MA, Baile CA (2007) Guggulsterone inhibits adipocyte differentiation and induces apoptosis in 3T3-L1 cells. Obesity (Silver Spring, Md), in pressGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Jeong-Yeh Yang
    • 1
  • Mary Anne Della-Fera
    • 1
  • Srujana Rayalam
    • 1
  • Clifton A. Baile
    • 1
    • 2
  1. 1.Department of Animal and Dairy Science, 444 Edgar L. Rhodes Center for Animal and Dairy ScienceUniversity of GeorgiaAthensUSA
  2. 2.Department of Foods and NutritionUniversity of GeorgiaAthensUSA

Personalised recommendations