7,8-hydroxy-2 '-deoxyguanosine/2 '-deoxiguanosine ratio determined in hydrolysates of brain DNA by ultrachromatrography coupled to tandem mass spectrometry

Data de publicació: 01/08/2017

Autors de IIS La Fe

María Isabel Torres Cuevas

Autor
Miguel Aupí Ferrando

Autor
Miguel Angel Asensi Miralles

Autor
Máximo Vento Torres

Autor
Ana Ortega Gutierrez

Autor
Justo Javier Escobar Cubiella

Autor

Grups d'Investigació

Grupo de Investigación Clínica y Traslacional en Cardiología

Investigador Principal

Esther Roselló Lletí
Perinatología

Leader

Máximo Vento Torres

Abstract

7,8-hydroxy-2'-deoxyguanosine (8-OHdG) is an abundant DNA lesion formed by oxidation of the nucleoside 2'-deoxyguanosine (2-dG) and one of the most studied and accepted oxidative stress biomarkers. 8-OHdG has a strong carcinogenic potential, and prolonged oxidative stress heightens pathological conditions and especially cancer risk. Our aim was to develop, validate and apply a reliable method to assess DNA oxidation in genomic cellular DNA of sensible target organs such as brain. A procedure to isolate and digest the DNA of brain tissue properly for further detection of 8-OHdG and 2-dG by Ultra Performance Liquid Chromatography tandem Mass Spectrometry (UPLC-MS/MS) was optimized. The UPLC-MS/MS was validated following the American Food and Drug Administration (FDA) Guidelines using mice pups' brain samples. To demonstrate the applicability of the UPLC-MS/MS method, 8-OHdG/2-dG ratio was determined in brain tissue of 1 day old newborn mice pups (P1) in a model of hypoxia pre-conditioning during fetal-to-neonatal transition. We found that hypoxia at birth (FiO(2) 0.14) and for 8 h thereafter induced lower levels of DNA oxidation in mice pups and rendered even protective against a postnatal asphyxia/reoxygenation insult compared with fetal to neonatal transition in room air. We conclude that the UPLC-MS/MS method developed has proven suitable for the analysis of DNA oxidation biomarker 8-OHdG/2-dG ratio in tissue samples from newborn mice pups. We aim to apply this method in future studies aiming to provide a deeper insight into the mechanisms of oxidation DNA caused during neonatal asphyxia and resuscitation.

Dades de la publicació

ISSN/ISSNe:: 0039-9140, 1873-3573
Tipus:: Article
Pàgines:: 97-102
DOI:: 10.1016/j.talanta.2017.03.072
PubMed:: 28501220
Factor d'Impacte:: 1,186 SCImago ℠

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Filiacions

Torres-Cuevas, I:: Instituto de Investigación. Neonatal research Group, Health Research Institute La Fe, Avenida Fernando Abril Martorell 106, 46026 Valencia, Spain

Instituto de Investigación. Hlth Res Inst La Fe, Neonatal Res Grp, Ave Fernando Abril Martorell 106, Valencia 46026, Spain
Aupi, M:: Department of Physiology, University of Valencia, Vicente Andrés Estellés s/n, 46100 Burjassot, Spain

Univ Valencia, Dept Physiol, Vicente Andres Estelles S-N, Burjassot 46100, Spain
Asensi, MA:: Department of Physiology, University of Valencia, Vicente Andrés Estellés s/n, 46100 Burjassot, Spain

Univ Valencia, Dept Physiol, Vicente Andres Estelles S-N, Burjassot 46100, Spain
Vento, M:: Instituto de Investigación. Neonatal research Group, Health Research Institute La Fe, Avenida Fernando Abril Martorell 106, 46026 Valencia, Spain

National Coordinator of the Spanish Maternal and Child Health and Developmental Network (Retic Red SAMID RD0016/0022), Spain

Instituto de Investigación. Hlth Res Inst La Fe, Neonatal Res Grp, Ave Fernando Abril Martorell 106, Valencia 46026, Spain

Natl Coordinator Spanish Maternal & Child Hlth &, Valencia, Spain
Ortega, A:: Department of Physiology, University of Valencia, Vicente Andrés Estellés s/n, 46100 Burjassot, Spain

Univ Valencia, Dept Physiol, Vicente Andres Estelles S-N, Burjassot 46100, Spain
Escobar, J:: Instituto de Investigación. Neonatal research Group, Health Research Institute La Fe, Avenida Fernando Abril Martorell 106, 46026 Valencia, Spain

Biotechnological transfer company Sabartech SL, Avenida Fernando Abril Martorell 106, 46026 Valencia, Spain

Instituto de Investigación. Hlth Res Inst La Fe, Neonatal Res Grp, Ave Fernando Abril Martorell 106, Valencia 46026, Spain

Biotechnol Transfer Co Sabartech SL, Ave Fernando Abril Martorell 106, Valencia 46026, Spain