Protocol

Free Radicals and Antioxidant Protocols

Volume 610 of the series Methods in Molecular Biology pp 245-261

Date:

Biomarkers of Oxidative Stress: Methods and Measures of Oxidative DNA Damage (COMET Assay) and Telomere Shortening

  • Muthuswamy BalasubramanyamAffiliated withDepartment of Cell and Molecular Biology, Madras Diabetes Research Foundation & Dr Mohan’s Diabetes Specialties Centre
  • , Antonysunil AdaikalakoteswariAffiliated withDepartment of Cell and Molecular Biology, Madras Diabetes Research Foundation & Dr Mohan’s Diabetes Specialties
  • , Zaheer SameermahmoodAffiliated withDepartment of Cell and Molecular Biology, Madras Diabetes Research Foundation & Dr Mohan’s Diabetes Specialties
  • , Viswanathan MohanAffiliated withMadras Diabetes Research Foundation & Dr Mohan’s Diabetes Specialties Centre

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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.

Key words

Oxidative stress telomere shortening comet assay DNA damage biomarker diabetes