title: Quantification of oxidative DNA lesions in tissues of Long-Evans Cinnamon rats by capillary high-performance liquid chromatography-tandem mass spectrometry coupled with stable isotope-dilution method.
abstract: The purpose of our study was to develop suitable methods to quantify oxidative DNA lesions in the setting of transition metal-related diseases. Transition metal-driven Fenton reactions constitute an important endogenous source of reactive oxygen species (ROS). In genetic diseases with accumulation of transition metal ions, excessive ROS production causes pathophysiological changes, including DNA damage. Wilson's disease is an autosomal recessive disorder with copper toxicosis due to deficiency of ATP7B protein needed for excreting copper into bile. The Long-Evans Cinnamon (LEC) rat bears a deletion in Atp7b gene and serves as an excellent model for hepatic Wilson's disease. We used a sensitive capillary liquid chromatography-electrospray-tandem mass spectrometry (LC-ESI-MS/MS/MS) method in conjunction with the stable isotope-dilution technique to quantify several types of oxidative DNA lesions in the liver and brain of LEC rats. These lesions included 5-formyl-2'-deoxyuridine, 5-hydroxymethyl-2'-deoxyuridine, and the 5'R and 5'S diastereomers of 8,5'-cyclo-2'-deoxyguanosine and 8,5'-cyclo-2'-deoxyadenosine. Moreover, the levels of these DNA lesions in the liver and brain increased with age and correlated with age-dependent regulation of the expression of DNA repair genes in LEC rats. These results provide significant new knowledge for better understanding the implications of oxidative DNA lesions in transition metal-induced diseases, such as Wilson's disease, as well as in aging and aging-related pathological conditions.
authors.affiliation: Department of Chemistry, University of California, Riverside, California 92521-0403, United States.
authors.forename: Jin
authors.initials: J
authors.lastname: Wang
authors.forename: Bifeng
authors.initials: B
authors.lastname: Yuan
authors.forename: Candace
authors.initials: C
authors.lastname: Guerrero
authors.forename: Ralf
authors.initials: R
authors.lastname: Bahde
authors.forename: Sanjeev
authors.initials: S
authors.lastname: Gupta
authors.forename: Yinsheng
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004249:DNA Damage; D004260:DNA Repair / Q000235:genetics; D003839:Deoxyadenosines / Q000737:chemistry / Q000378:metabolism; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D003857:Deoxyuridine / Q000031:analogs & derivatives / Q000378:metabolism; D005786:Gene Expression Regulation; D007554:Isotopes; D008099:Liver / Q000166:cytology / Q000378:metabolism; D010084:Oxidation-Reduction; D051381:Rats; D020332:Rats, Inbred LEC; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013936:Thymidine / Q000031:analogs & derivatives / Q000378:metabolism; D013997:Time Factors
chemical_list: D003839:Deoxyadenosines; D007554:Isotopes; D017382:Reactive Oxygen Species; C050888:8,5'-cyclo-2'-deoxyguanosine; C056752:8,5'-cyclo-2'-deoxyadenosine; C061519:5-formyl-2'-deoxyuridine; C018080:5-hydroxymethyl-2'-deoxyuridine; D003849:Deoxyguanosine; D013936:Thymidine; D003857:Deoxyuridine