Abstract
2-Deoxy-d-glucose (2-DG) and dehydroepiandrosterone (DHEA) have been hypothesized to extend lifespan via mimicking calorie restriction (CR). Activation of sirtuins has been proposed to contribute to life extension of CR by increasing intercellular levels of NAD+ in several organisms. However, it is unclear whether 2-DG and DHEA may affect intracellular NAD+ levels and human sirtuin 1 (SIRT1) activities. Here, using human fibroblast Hs68 cells we showed that 2-DG increased intracellular NAD+ levels in both time- and concentration-dependent manners. 2-DG also dose-dependently increased SIRT1 activities and the lifespan (measured as the cumulated growth curve of population doubling levels) of Hs68 cells. In contrast, DHEA at non-cytotoxic concentrations (≤50 μM) did not significantly affect NAD+ levels, SIRT1 activities or the lifespan of Hs68 cells. These results suggest that 2-DG extends the lifespan of Hs68 cells by increased NAD+ levels and SIRT1 activities, and that 2-DG has a potential as a CR mimetic.
Similar content being viewed by others
References
Aljada A, Dong L, Mousa SA (2010) Sirtuin-targeting drugs: mechanisms of action and potential therapeutic applications. Curr Opin Investig Drugs 11:1158–1168
Anderson RM, Bitterman KJ, Wood JG, Medvedik O, Cohen H, Lin SS, Manchester JK, Gordon JI, Sinclair DA (2002) Manipulation of a nuclear NAD+ salvage pathway delays aging without altering steady-state NAD+ levels. J Biol Chem 277:18881–18890. doi:10.1074/jbc.M111773200
Arnold JT (2009) DHEA metabolism in prostate: for better or worse? Mol Cell Endocrinol 301:83–88. doi:10.1016/j.mce.2008.10.019
Barbieri M, Bonafè M, Franceschi C, Paolisso G (2003) Insulin/IGF-I-signaling pathway: an evolutionarily conserved mechanism of longevity from yeast to humans. Am J Physiol Endocrinol Metab 285:E1064–E1071. doi:10.1152/ajpendo.00296.2003
Bonkowski MS, Rocha JS, Masternak MM, Al Regaiey KA, Bartke A (2006) Targeted disruption of growth hormone receptor interferes with the beneficial actions of calorie restriction. Proc Natl Acad Sci USA 103:7901–7905. doi:10.1073/pnas.0600161103
Campisi J (2001) From cells to organisms: can we learn about aging from cells in culture? Exp Gerontol 36:607–618. doi:10.1016/S0531-5565(00)00230-8
Chen D, Guarente L (2007) SIR2: a potential target for calorie restriction mimetics. Trends Mol Med 13:64–71. doi:10.1016/j.molmed.2006.12.004
Donmez G, Guarente L (2010) Aging and disease: connections to sirtuins. Aging Cell 9:285–290. doi:10.1111/j.1474-9726.2010.00548.x
Fulco M, Cen Y, Zhao P, Hoffman EP, McBurney MW, Sauve AA, Sartorelli V (2008) Glucose restriction inhibits skeletal myoblast differentiation by activating SIRT1 through AMPK-mediated regulation of Nampt. Dev Cell 14:661–673. doi:10.1016/j.devcel.2008.02.004
Ghosh HS (2008) The anti-aging, metabolism potential of SIRT1. Curr Opin Investig Drugs 9:1095–1102
Haigis MC, Sinclair DA (2010) Mammalian sirtuins: biological insights and disease relevance. Annu Rev Pathol 5:253–295. doi:10.1146/annurev.pathol.4.110807.092250
Hashimoto T, Horikawa M, Nomura T, Sakamoto K (2010) Nicotinamide adenine dinucleotide extends the lifespan of Caenorhabditis elegans mediated by sir-2.1 and daf-16. Biogerontology 11:31–43. doi:10.1007/s10522-009-9225-3
Hayflick L, Moorehead PS (1961) The serial cultivation of human diploid cell strains. Exp Cell Res 25:585–621
Ho HY, Cheng ML, Lu FJ, Chou YH, Stern A, Liang CM, Chiu DT (2000) Enhanced oxidative stress and accelerated cellular senescence in glucose-6-phosphate dehydrogenase (G6PD)-deficient human fibroblasts. Free Radic Biol Med 29:156–169. doi:10.1016/S0891-5849(00)00331-2
Ingram DK, Zhu M, Mamczarz J, Zou S, Lane MA, Roth GS, deCabo R (2006) Calorie restriction mimetics: an emerging research field. Aging Cell 5:97–108. doi:10.1111/j.1474-9726.2006.00202.x
Kang HT, Hwang ES (2006) 2-Deoxyglucose: an anticancer and antiviral therapeutic, but not any more a low glucose mimetic. Life Sci 78:1392–1399. doi:10.1016/j.lfs.2005.07.001
Kyrylenko S, Baniahmad A (2010) Sirtuin family: a link to metabolic signaling and senescence. Curr Med Chem 17:2921–2932. doi:10.2174/092986710792065009
Lane MA, Mattison J, Ingram DK, Roth GS (1998) 2-Deoxyglucose feeding in rats mimics physiological effects of caloric restriction. J Anti-Aging Med 1:327–337. doi:10.1089/rej.1.1998.1.327
Lane MA, Roth GS, Ingram DK (2007) Caloric restriction mimetics: a novel approach for biogerontology. Methods Mol Biol 371:143–149. doi:10.1007/978-1-59745-361-5_11
Lim CS, Potts M, Helm RF (2006) Nicotinamide extends the replicative life span of primary human cells. Mech Ageing Dev 127:511–514. doi:10.1016/j.mad.2006.02.001
Lin SJ, Ford E, Haigis M, Liszt G, Guarente L (2004) Calorie restriction extends yeast life span by lowering the level of NADH. Genes Dev 18:12–16. doi:10.1101/gad.1164804
Lu SP, Lin SJ (2010) Regulation of yeast sirtuins by NAD(+) metabolism and calorie restriction. Biochim Biophys Acta 1804:1567–1575. doi:10.1016/j.bbapap.2009.09.030
Matuoka K, Chen KY, Takenawa T (2001) Rapid reversion of aging phenotypes by nicotinamide through possible modulation of histone acetylation. Cell Mol Life Sci 58:2108–2116. doi:10.1007/PL00000840
McFarland GA, Holliday R (1994) Retardation of the senescence of cultured human diploid fibroblasts by carnosine. Exp Cell Res 212:167–175. doi:10.1006/excr.1994.1132
Minor RK, Smith DL Jr, Sossong AM, Kaushik S, Poosala S, Spangler EL, Roth GS, Lane M, Allison DB, de Cabo R, Ingram DK, Mattison JA (2010) Chronic ingestion of 2-deoxy-d-glucose induces cardiac vacuolization and increases mortality in rats. Toxicol Appl Pharmacol 243:332–339. doi:10.1016/j.taap.2009.11.025
Nair KS, Rizza RA, O’Brien P, Dhatariya K, Short KR, Nehra A, Vittone JL, Klee GG, Basu A, Basu R, Cobelli C, Toffolo G, Dalla Man C, Tindall DJ, Melton LJ 3rd, Smith GE, Khosla S, Jensen MD (2006) DHEA in elderly women and DHEA or testosterone in elderly men. N Engl J Med 355:1647–1659
Pelicano H, Martin DS, Xu RH, Huang P (2006) Glycolysis inhibition for anticancer treatment. Oncogene 25:4633–4646. doi:10.1038/sj.onc.1209597
Pugh DP, Oberley TD, Weindruch R (1999) Dietary intervention at middle age: caloric restriction but no dehydroepiandrosterone sulfate increases lifespan and lifetime cancer incidence in mice. Cancer Res 59:1642–1648
Sánchez J, Pérez-Heredia F, Priego T, Portillo MP, Zamora S, Garaulet M, Palou A (2008) Dehydroepiandrosterone prevents age-associated alterations, increasing insulin sensitivity. J Nutr Biochem 19:809–818. doi:10.1016/j.jnutbio.2007.10.005
Schmidt MT, Smith BC, Jackson MD, Denu JM (2004) Coenzyme specificity of sir2 protein deacetylases: implications for physiological regulation. J Biol Chem 279:40122–40129. doi:10.1074/jbc.M407484200
Singh D, Banerji AK, Dwarakanath BS, Tripathi RP, Gupta JP, Mathew TL, Ravindranath T, Jain V (2005) Optimizing cancer radiotherapy with 2-deoxy-d-glucose dose escalation studies in patients with glioblastoma multiforme. Strahlenther Onkol 181:507–514. doi:10.1007/s00066-005-1320-z
Srivastava DK, Bernhard SA (1987) Biophysical chemistry of metabolic reaction sequences in concentrated enzyme solution and in the cell. Annu Rev Biophys Biophys Chem 16:175–204
Swierczyński J, Słomińska E, Smoleński RT, Mayer D (2001) Increase in NAD but not ATP and GTP concentrations in rat liver by dehydroepiandrosterone feeding. Pol J Pharmacol 53:125–130
Williams JR (2000) The effects of dehydroepiandrosterone on carcinogenesis, obesity, the immune system and aging. Lipids 35:325–331. doi:10.1007/s11745-000-0529-7
Wood JG, Rogina B, Lavu S, Howitz K, Helfand SL, Tatar M, Sinclair D (2004) Sirtuin activators mimic caloric restriction and delay ageing in metazoans. Nature 430:686–689. doi:10.1038/nature02789
Yang NC, Hu ML (2004) A fluorimetric method using fluorescein di-beta-D-galactopyranoside for quantifying the senescence-associated beta-galactosidase activity in human foreskin fibroblast Hs68 cells. Anal Biochem 325:337–343. doi:10.1016/j.ab.2003.11.012
Yang NC, Hu ML (2005) The limitations and validities of senescence associated-beta-galactosidase activity as an aging marker for human foreskin fibroblast Hs68 cells. Exp Gerontol 40:813–819. doi:10.1016/j.exger.2005.07.011
Yang NC, Jeng KC, Ho WM, Chou SJ, Hu ML (2000) DHEA inhibits cell growth and induces apoptosis in BV-2 cells and the effects are inversely associated with glucose concentration in the medium. J Steroid Biochem Mol Biol 75:159–166. doi:10.1016/S0960-0760(00)00180-1
Yang NC, Jeng KC, Ho WM, Hu ML (2002) ATP depletion is an important factor in DHEA-induced growth inhibition and apoptosis in BV-2 cells. Life Sci 70:1979–1988. doi:10.1016/S0024-3205(01)01542-9
Zerez CR, Lee SJ, Tanaka KR (1987) Spectrophotometric determination of oxidized and reduced pyridine nucleotides in erythrocytes using a single extraction procedure. Anal Biochem 164:367–373. doi:10.1016/0003-2697(87)90506-9
Acknowledgments
This research was supported by grants from the National Science Council (NSC-98-2313-B-235-001).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yang, NC., Song, TY., Chen, MY. et al. Effects of 2-deoxyglucose and dehydroepiandrosterone on intracellular NAD+ level, SIRT1 activity and replicative lifespan of human Hs68 cells. Biogerontology 12, 527–536 (2011). https://doi.org/10.1007/s10522-011-9342-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10522-011-9342-7