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HPLC Measurement of the DNA Oxidation Biomarker, 8-oxo-7,8-dihydro-2’-deoxyguanosine, in Cultured Cells and Animal Tissues
Published on: August 1, 2015
Levels of 8-hydroxy-2'-deoxyguanosine in cellular DNA from 12 tissues of young and old Sprague-Dawley rats
1Institute of Pharmacology and Toxicology, Veterinary University of Vienna, Veterinärplatz 1, A-1210 Vienna, Austria. ivo.scherold@vu-wien.ac.at
Abstract:
The age dependent increase of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) reported in DNA of organs of old rats appears to vary with the strain, age and sex of the animals used for the investigation. Here we report on 8-OH-dG concentrations in the cellular DNA of 12 tissues of male Sprague-Dawley rats aged 5 or 30 months and kept under standard conditions throughout their lives until being killed. DNA from frozen organs was isolated using a Qiagen DNA purification kit. Following digestion (nuclease P1, alkaline nuclease) hydrolysed DNA was applied onto a HPLC column; native nucleosides were monitored at 254 nm and 8-OH-dG by electrochemical detection. 8-OH-dG levels in organs of young rats ranged between 10 and 90 micromol/mol deoxyguanosine (dG). Highest levels (micromol 8-OH-dG /mol dG) were detected in the oesophagus (90), muscle (61), brain (65), liver (59), spleen (57), and testicles (63). 8-OH-dG in DNA from kidney, lung, heart, small and large intestine ranged between 28 and 38 micromol/mol dG. Lowest amounts were found in the glandular stomach (10). DNA of old rats generally contained higher 8-OH-dG levels with significant increases in liver (186%) and kidney (372%); other organs showed no significant decreases (spleen, brain, testicles) or increases up to 69% (heart). These findings are discussed in the context with previously published data on 8-OH-dG levels in organs from young and old rats.
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