Abstract
Oxidative stress is fast becoming the nutritional and medical buzzword for the twenty-first century. The theoretical importance of oxidative stress in diabetes is highlighted by its potential double impact on metabolic dysfunction on one hand and the vascular system on the other hand. The new concept of oxidative stress, being an important trigger in the onset and progression of diabetes and its complications, emphasizes the need for measurement of markers of oxidation to assess the degree of oxidative stress. While we have been routinely measuring biomarkers in our molecular epidemiology projects, here we discuss the utility of two assays, (a) DNA damage assessment by COMET measurement and (b) telomere length measurement. As DNA damage is efficiently repaired by cellular enzymes, its measurement gives a snapshot view of the level of oxidative stress. The protocol allows for measurement of oxidative DNA damage (FPG-sensitive DNA strand breaks). Telomere length measured by Southern blotting technique allows one to estimate the chronic burden of oxidative stress at the molecular level and is now considered as biomarker of biological aging.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ceriello, A. and Motz, E. (2004) Is oxidative stress the pathogenic mechanism underlying insulin resistance, diabetes, and cardiovascular disease? The common soil hypothesis revisited. Arterioscler. Thromb. Vasc. Biol. 24, 816–823.
Balasubramanyam, M. (2006) Antioxidants and cardiovascular disease – Where do we stand? Asian J. Diabetol. 8, 1–5.
Evans, J.L., Maddux, B.A., and Goldfine, I.D. (2005) The molecular basis for oxidative stress-induced insulin resistance. Antioxid. Redox. Signal 7, 1040–1052.
Evans, M.D. and Cooke, M.S. (2004) Factors contributing to the outcome of oxidative damage to nucleic acids. Bioessays 26, 533–542.
Singh, N.P., McCoy, M.T., Tice, R.R., and Schneider, E.L. (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell. Res. 175, 184–191.
Collins, A.R., Raslova, K., Somorovska, M., Petrovska, H., Ondrusova, A., Vohnout, B., Fabry, R., and Dusinska, M. (1998) DNA damage in diabetes: correlation with a clinical marker. Free Radic. Biol. Med. 25, 373–377.
Adaikalakoteswari, A., Rema, M., Mohan, V., and Balasubramanyam, M. (2007). Oxidative DNA damage and augmentation of Poly (ADP-ribose) polymerase/Nuclear factor-kappa B signaling in patients with Type 2 diabetes and microangiopathy. Int. J. Biochem. Cell Biol. 39,1673–1684.
Collins, A.R. (2004) The comet assay for DNA damage and repair: principles, applications, and limitations. Mol. Biotechnol. 26, 249–261.
Cadet, J., Bellon, S., Berger, M., Bourdat, A.G., Douki, T., Duarte, V., Frelon, S., Gasparutto, D., Muller, E., Ravanat, J.L., and Sauvaigo, S. (2002) Recent aspects of oxidative DNA damage: guanine lesions, measurement and substrate specificity of DNA repair glycosylases. Biol. Chem. 383, 933–943.
Kawanishi, S. and Oikawa, S. (2004) Mechanism of telomere shortening by oxidative stress. Ann. NY Acad. Sci. 1019, 278–284.
Von Zglinicki, T. and Martin-Ruiz, C.M. (2005) Telomeres as biomarkers for ageing and age-related diseases. Curr. Mol. Med. 5, 197–203.
Samani, N.J., Boultby, R., Butler, R., Thompson, J.R., and Goodall, A.H. (2001) Telomere shortening in atherosclerosis. Lancet 358, 472– 473.
Jeanclos, E., Schork, N.J., Kyvik, K.O., Kimura, M., Skurnick, J.H., and Aviv, A. (2000) Telomere length inversely correlates with pulse pressure and is highly familial. Hypertension 36, 195–200.
Adaikalakoteswari, A., Balasubramanyam, M., and Mohan, V. (2005) Telomere shortening occurs in Asian Indian type 2 diabetic patients. Diabet. Med. 22, 1151–1156.
Adaikalakoteswari, A., Balasubramanyam, M., Ravikumar, R., Deepa, R., and Mohan, V. (2007) Association of telomere shortening with impaired glucose tolerance and diabetic macroangiopathy. Atherosclerosis 195, 83–89.
Brouilette, S.W., Moore, J.S., McMahon, A.D., Thompson, J.R., Ford, I., Shepherd, J., et al. (2007) West of Scotland Coronary Prevention Study Group. Telomere length, risk of coronary heart disease, and statin treatment in the West of Scotland Primary Prevention Study: a nested case-control study. Lancet 369, 107–114.
Acknowledgments
This work was supported by research grants from the Department of Science and Technology (DST & DST-FIST), the Department of Biotechnology (DBT), and the Indian Council of Medical Research (ICMR), Government of India.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Balasubramanyam, M., Adaikalakoteswari, A., Sameermahmood, Z., Mohan, V. (2010). Biomarkers of Oxidative Stress: Methods and Measures of Oxidative DNA Damage (COMET Assay) and Telomere Shortening. In: Uppu, R., Murthy, S., Pryor, W., Parinandi, N. (eds) Free Radicals and Antioxidant Protocols. Methods in Molecular Biology, vol 610. Humana Press. https://doi.org/10.1007/978-1-60327-029-8_15
Download citation
DOI: https://doi.org/10.1007/978-1-60327-029-8_15
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-58829-710-5
Online ISBN: 978-1-60327-029-8
eBook Packages: Springer Protocols