AGE

, Volume 34, Issue 1, pp 95–109 | Cite as

MnSOD activity regulates hydroxytyrosol-induced extension of chronological lifespan

  • Ehab H. Sarsour
  • Maneesh G. Kumar
  • Amanda L. Kalen
  • Monali Goswami
  • Garry R. Buettner
  • Prabhat C. Goswami
Article

Abstract

Chronological lifespan (CLS) is defined as the duration of quiescence in which normal cells retain the capacity to reenter the proliferative cycle. This study investigates whether hydroxytyrosol (HT), a naturally occurring polyphenol found in olives, extends CLS in normal human fibroblasts (NHFs). Quiescent NHFs cultured for a long duration (30–60 days) lose their capacity to repopulate. Approximately 60% of these cells exit the cell cycle permanently; a significant increase in the doubling time of the cell population was observed. CLS was extended in quiescent NHFs that were cultured in the presence of HT for 30–60 days. HT-induced extension of CLS was associated with an approximately 3-fold increase in manganese superoxide dismutase (MnSOD) activity while there was no change in copper–zinc superoxide dismutase, catalase, or glutathione peroxidase protein levels. Quiescent NHFs overexpressing a dominant-negative mutant form of MnSOD failed to extend CLS. HT suppressed age-associated increase in mitochondrial ROS levels. Results from spectroscopy assays indicate that HT in the presence of peroxidases can undergo catechol–semiquinone–quinone redox cycling generating superoxide, which in a cellular context can activate the antioxidant system, e.g., MnSOD expression. These results demonstrate that HT extends CLS by increasing MnSOD activity and decreasing age-associated mitochondrial reactive oxygen species accumulation.

Keywords

Chronological lifespan Ageing Manganese superoxide dismutase Quiescence Hydroxytyrosol Mitochondria 

Abbreviations

AdmMnSOD

Adenoviruses carrying a dominant-negative form of human MnSOD cDNA

CLS

Chronological lifespan

CuZnSOD

Copper–zinc superoxide dismutase

EcSOD

Extracellular superoxide dismutase

EPR

Electron paramagnetic resonance

GPx

Glutathione peroxidase

HT

Hydroxytyrosol

MnSOD

Manganese superoxide dismutase

MOI

Multiplicity of infection

NHFs

Normal human fibroblasts

PI

Propidium iodide

ROS

Reactive oxygen species

Notes

Acknowledgments

We thank The University of Iowa EPR and Flow Cytometry Cores for assisting with EPR spectroscopy and flow cytometry assays. Funding from NIH CA 111365 and McCord Research foundation supported this work.

Supplementary material

11357_2011_9223_MOESM1_ESM.pdf (300 kb)
ESM 1 (PDF 300 kb)

References

  1. Allsopp RC, Chang E et al (1995) Telomere shortening is associated with cell division in vitro and in vivo. Exp Cell Res 220:194–200. doi: 10.1006/excr.1995.1306 PubMedCrossRefGoogle Scholar
  2. Bachur NR, Gordon SL, Gee MV (1978) A general mechanism for microsomal activation of quinone anticancer agents to free radicals. Cancer Res 38:1745–1750PubMedGoogle Scholar
  3. Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287PubMedCrossRefGoogle Scholar
  4. Bodnar AG, Ouellette M et al (1998) Extension of life-span by introduction of telomerase into normal human cells. Science 279:349–352PubMedCrossRefGoogle Scholar
  5. Buettner GR, Ng CF et al (2006) A new paradigm: manganese superoxide dismutase influences the production of H2O2 in cells and thereby their biological state. Free Radic Biol Med 41:1338–1350. doi: 10.1016/j.freeradbiomed.2006.07.015 PubMedCrossRefGoogle Scholar
  6. Cornwell DG, Ma J (2008) Nutritional benefit of olive oil: the biological effects of hydroxytyrosol and its arylating quinone adducts. J Agric Food Chem 56:8774–8786. doi: 10.1021/jf8015877 PubMedCrossRefGoogle Scholar
  7. D’Angelo S, Ingrosso D et al (2005) Hydroxytyrosol, a natural antioxidant from olive oil, prevents protein damage induced by long-wave ultraviolet radiation in melanoma cells. Free Radic Biol Med 38:908–919. doi: 10.1016/j.freeradbiomed.2004.12.015 PubMedCrossRefGoogle Scholar
  8. De Lucia M, Panzella L et al (2006) Oxidative chemistry of the natural antioxidant hydroxytyrosol: hydrogen peroxide-dependent hydroxylation and hydroxyquinone/o-quinone coupling pathways. Tetrahedron 62:1273–1278. doi: 10.1016/j.tet.2005.10.055 CrossRefGoogle Scholar
  9. Eyer P (1991) Effects of superoxide dismutase on the autoxidation of 1,4-hydroquinone. Chem Biol Interact 80:159–176PubMedCrossRefGoogle Scholar
  10. Fabiani R, Rosignoli P et al (2008) Oxidative DNA damage is prevented by extracts of olive oil, hydroxytyrosol, and other olive phenolic compounds in human blood mononuclear cells and HL60 cells. J Nutr 138:1411–1416PubMedGoogle Scholar
  11. Fabrizio P, Pletcher SD et al (2004) Chronological aging-independent replicative life span regulation by Msn2/Msn4 and Sod2 in Saccharomyces cerevisiae. FEBS Lett 557:136–142PubMedCrossRefGoogle Scholar
  12. Goldstein S, Singal DP (1974) Senescence of cultured human fibroblasts: mitotic versus metabolic time. Exp Cell Res 88:359–364PubMedCrossRefGoogle Scholar
  13. Hamden K, Allouche N, Damak M, Elfeki A (2009) Hypoglycemic and antioxidant effects of phenolic extracts and purified hydroxytyrosol from olive mill waste in vitro and in rats. Chem Biol Interact 180:421–432. doi: 10.1016/j.cbi.2009.04.002 PubMedCrossRefGoogle Scholar
  14. Harris N, Costa V et al (2003) Mnsod overexpression extends the yeast chronological (G(0)) life span but acts independently of Sir2p histone deacetylase to shorten the replicative life span of dividing cells. Free Radic Biol Med 34:1599–1606.PubMedCrossRefGoogle Scholar
  15. Hayflick L, Moorhead PS (1961) The serial cultivation of human diploid cell strains. Exp Cell Res 25:585–621CrossRefGoogle Scholar
  16. Henderson ER, Larson DD (1991) Telomeres—what’s new at the end? Curr Opin Genet Dev 1:538–543PubMedCrossRefGoogle Scholar
  17. Jemai H, Fki I et al (2008) Lipid-lowering and antioxidant effects of hydroxytyrosol and its triacetylated derivative recovered from olive tree leaves in cholesterol-fed rats. J Agric Food Chem 56:2630–2636. doi: 10.1021/jf072589s PubMedCrossRefGoogle Scholar
  18. Jemai H, El Feki A, Sayadi S (2009) Antidiabetic and antioxidant effects of hydroxytyrosol and oleuropein from olive leaves in alloxan-diabetic rats. J Agric Food Chem 57:8798–8804. doi: 10.1021/jf901280r PubMedCrossRefGoogle Scholar
  19. Manna C, Della Ragione F et al (1999) Biological effects of hydroxytyrosol, a polyphenol from olive oil endowed with antioxidant activity. Adv Exp Med Biol 472:115–130PubMedGoogle Scholar
  20. Menon SG, Sarsour EH et al (2003) Redox regulation of the G1 to S phase transition in the mouse embryo fibroblast cell cycle. Cancer Res 63:2109–2117PubMedGoogle Scholar
  21. Mitra K, Wunder C et al (2009) A hyperfused mitochondrial state achieved at G1-S regulates cyclin E buildup and entry into S phase. Proc Natl Acad Sci USA 106:11960–11965. doi: 10.1073/pnas.0904875106 PubMedCrossRefGoogle Scholar
  22. Mukherjee S, Lekli I et al (2009) Expression of the longevity proteins by both red and white wines and their cardioprotective components, resveratrol, tyrosol, and hydroxytyrosol. Free Radic Biol Med 46:573–578. doi: 10.1016/j.freeradbiomed.2008.11.005 PubMedCrossRefGoogle Scholar
  23. Munro J, Steeghs K et al (2001) Human fibroblast replicative senescence can occur in the absence of extensive cell division and short telomeres. Oncogene 20:3541–3552. doi: 10.1038/sj.onc.1204460 PubMedCrossRefGoogle Scholar
  24. O’Dowd Y, Driss F et al (2004) Antioxidant effect of hydroxytyrosol, a polyphenol from olive oil: scavenging of hydrogen peroxide but not superoxide anion produced by human neutrophils. Biochem Pharmacol 68:2003–2008. doi: 10.1016/j.bcp.2004.06.023 PubMedCrossRefGoogle Scholar
  25. Orr WC, Mockett RJ, Benes JJ, Sohal RS (2003) Effects of overexpression of copper-zinc and manganese superoxide dismutases, catalase, and thioredoxin reductase genes on longevity in Drosophila melanogaster. J Biol Chem 278:26418–26422. doi: 10.1074/jbc.M303095200 PubMedCrossRefGoogle Scholar
  26. Rodriguez G, Lama A et al (2009) 3,4-Dihydroxyphenylglycol (DHPG): an important phenolic compound present in natural table olives. J Agric Food Chem 57:6298–6304. doi: 10.1021/jf803512r PubMedCrossRefGoogle Scholar
  27. Sarsour EH, Agarwal M et al (2005) Manganese superoxide dismutase protects the proliferative capacity of confluent normal human fibroblasts. J Biol Chem 280:18033–18041. doi: 10.1074/jbc.M501939200 PubMedCrossRefGoogle Scholar
  28. Sarsour EH, Venkataraman S et al (2008) Manganese superoxide dismutase activity regulates transitions between quiescent and proliferative growth. Aging Cell 7:405–417. doi: 10.1111/j.1474-9726.2008.00384.x PubMedCrossRefGoogle Scholar
  29. Sarsour EH, Goswami M, Kalen AL, Goswami PC (2010) MnSOD activity protects mitochondrial morphology of quiescent fibroblasts from age associated abnormalities. Mitochondrion 10:342–349. doi: 10.1016/j.mito.2010.02.004 PubMedCrossRefGoogle Scholar
  30. Schriner SE, Linford NJ et al (2005) Extension of murine life span by overexpression of catalase targeted to mitochondria. Science 308:1909–1911. doi: 10.1126/science.1106653 PubMedCrossRefGoogle Scholar
  31. Sitte N, Saretzki G, von Zglinicki T (1998) Accelerated telomere shortening in fibroblasts after extended periods of confluency. Free Radic Biol Med 24:885–893PubMedCrossRefGoogle Scholar
  32. Sohal RS, Sohal BH, Orr WC (1995) Mitochondrial superoxide and hydrogen peroxide generation, protein oxidative damage, and longevity in different species of flies. Free Radic Biol Med 19:499–504PubMedCrossRefGoogle Scholar
  33. Song Y, Buettner GR (2010) Thermodynamic and kinetic considerations for the reaction of semiquinone radicals to form superoxide and hydrogen peroxide. Free Radic Biol Med. doi: 10.1016/j.freeradbiomed.2010.05.009 Google Scholar
  34. Song Y, Wagner BA, Lehmler HJ, Buettner GR (2008) Semiquinone radicals from oxygenated polychlorinated biphenyls: electron paramagnetic resonance studies. Chem Res Toxicol 21:1359–1367. doi: 10.1021/tx8000175 PubMedCrossRefGoogle Scholar
  35. Song Y, Wagner BA et al (2009) Nonenzymatic displacement of chlorine and formation of free radicals upon the reaction of glutathione with PCB quinones. Proc Natl Acad Sci USA 106:9725–9730. doi: 10.1073/pnas.0810352106 PubMedCrossRefGoogle Scholar
  36. Sun J, Folk D, Bradley TJ, Tower J (2002) Induced overexpression of mitochondrial Mn-superoxide dismutase extends the life span of adult Drosophila melanogaster. Genetics 161:661–672PubMedGoogle Scholar
  37. Taub J, Lau JF et al (1999) A cytosolic catalase is needed to extend adult lifespan in C. elegans daf-C and clk-1 mutants. Nature 399:162–166. doi: 10.1038/20208 PubMedCrossRefGoogle Scholar
  38. Vaziri H, Benchimol S (1998) Reconstitution of telomerase activity in normal human cells leads to elongation of telomeres and extended replicative life span. Curr Biol 8:279–282PubMedCrossRefGoogle Scholar
  39. Venkatesha VA, Venkataraman S et al (2008) Catalase ameliorates polychlorinated biphenyl-induced cytotoxicity in nonmalignant human breast epithelial cells. Free Radic Biol Med 45:1094–1102. doi: 10.1016/j.freeradbiomed.2008.07.007 PubMedCrossRefGoogle Scholar
  40. Venkatesha VA, Kalen AL, Sarsour EH, Goswami PC (2010) PCB-153 exposure coordinates cell cycle progression and cellular metabolism in human mammary epithelial cells. Toxicol Lett 196:110–116. doi: 10.1016/j.toxlet.2010.04.005 PubMedCrossRefGoogle Scholar
  41. Visioli F, Bellomo G, Galli C (1998) Free radical-scavenging properties of olive oil polyphenols. Biochem Biophys Res Commun 247:60–64. doi: 10.1006/bbrc.1998.8735 PubMedCrossRefGoogle Scholar
  42. Vogna D, Pezzella A et al (2003) Oxidative chemistry of hydroxytyrosol: isolation and characterisation of novel methanooxocinobenzodioxinone derivatives. Tetrahedron Lett 44:8289–8292. doi: 10.1016/j.tetlet.2003.09.066 CrossRefGoogle Scholar
  43. Wang C, Li Z et al (2006) Cyclin D1 repression of nuclear respiratory factor 1 integrates nuclear DNA synthesis and mitochondrial function. Proc Natl Acad Sci USA 103:11567–11572. doi: 10.1073/pnas.0603363103 PubMedCrossRefGoogle Scholar
  44. Wright WE, Brasiskyte D, Piatyszek MA, Shay JW (1996) Experimental elongation of telomeres extends the lifespan of immortal × normal cell hybrids. EMBO J 15:1734–1741PubMedGoogle Scholar
  45. Yamada K, Ogawa H et al (2009) Mechanism of the antiviral effect of hydroxytyrosol on influenza virus appears to involve morphological change of the virus. Antivir Res 83:35–44. doi: 10.1016/j.antiviral.2009.03.002 PubMedCrossRefGoogle Scholar
  46. Zhang YP, Smith BJ, Oberley LW (2006) Enzymatic activity is necessary for the tumor-suppressive effects of MnSOD. Antioxid Redox Signal 8:1283–1293PubMedCrossRefGoogle Scholar

Copyright information

© American Aging Association 2011

Authors and Affiliations

  • Ehab H. Sarsour
    • 1
  • Maneesh G. Kumar
    • 1
  • Amanda L. Kalen
    • 1
  • Monali Goswami
    • 1
  • Garry R. Buettner
    • 1
  • Prabhat C. Goswami
    • 1
  1. 1.Free Radical and Radiation Biology Division, Department of Radiation OncologyThe University of IowaIowa CityUSA

Personalised recommendations