Quantification of Oxidative DNA Lesions in Tissues of Long-Evans Cinnamon Rats by Capillary High-Performance Liquid Chromatography鈭扵andem Mass Spectrometry Coupled with Stable Isotope-Dilution Method

详细信息    查看全文

• 作者：Jin Wang ; Bifeng Yuan ; Candace Guerrero ; Ralf Bahde ; Sanjeev Gupta ; Yinsheng Wang
• 刊名：Analytical Chemistry
• 出版年：2011
• 出版时间：March 15, 2011
• 年：2011
• 卷：83
• 期：6
• 页码：2201-2209
• 全文大小：988K
• 年卷期：v.83,no.6(March 15, 2011)
• ISSN：1520-6882

文摘

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鈥檚 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鈥檚 disease. We used a sensitive capillary liquid chromatography鈭抏lectrospray-tandem mass spectrometry (LC鈭扙SI-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鈥?deoxyuridine, 5-hydroxymethyl-2鈥?deoxyuridine, and the 5鈥?i>R and 5鈥?i>S diastereomers of 8,5鈥?cyclo-2鈥?deoxyguanosine and 8,5鈥?cyclo-2鈥?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鈥檚 disease, as well as in aging and aging-related pathological conditions.