8-Oxo-7,8-dihydroguanine and 8-oxo-7,8-dihydro-2'-deoxyguanosine concentrations in various human body fluids: implications for their measurement and interpretation
8-Oxo-7,8-dihydroguanine and 8-oxo-7,8-dihydro-2'-deoxyguanosine concentrations in various human body fluids: implications for their measurement and interpretation
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DOI：
10.1007/s00204-014-1255-1
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发表时间：
2015-02-01
影响因子：
6.1
通讯作者：
Chao, Mu-Rong
中科院分区：
文献类型：
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作者：
Hu, Chiung-Wen;Cooke, Marcus S.;Chao, Mu-Rong
8-氧-7,8-二氢-2'-脱氧鸟苷（8-oxodGuo）是DNA氧化损伤研究最多的产物，与衰老、癌症和一些退行性疾病的发展有关。在这里，我们提出了第一个液相色谱-串联质谱方法，可以同时测量其在血浆和唾液中的修复产物，即8-氧-7,8-二氢鸟嘌呤（8-氧瓜）和8-氧郭。利用这种方法，我们研究了氧化损伤DNA修复产物在细胞区室和生物基质之间的潜在运输机制。同时采集57名健康受试者的血浆、唾液和尿液样本。对各种脱蛋白方法进行了评价，并根据对回收率和色谱的影响，分别选择了血浆和唾液样品的沉淀剂乙腈和氢氧化钠-甲醇。血浆中8-oxoGua和8-oxodGuo的平均基线浓度分别为0.21和0.016 ng/mL，唾液中8-oxoGua和8-oxodGuo的平均基线浓度分别为0.85和0.010 ng/mL。唾液中8-oxoGua的浓度相对较高，其浓度因子（CF，唾液与血浆的浓度比）为4，而8-oxodGuo的浓度因子为0.6，这表明8-oxoGua在血浆中可能是主动转运到唾液中的，而8-oxodGuo主要依赖于被动扩散。8-氧瓜和8-氧果果在尿液和血浆浓度之间存在良好的相关性，这表明除了尿液之外，血液也是合适的基质。尿中8-oxoGua和8-oxodGuo只有在不校正尿肌酐浓度的情况下才存在显著相关性，这就提出了尿肌酐是否适用于调整8-oxoGua浓度的问题。
8-Oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) is the most investigated product of oxidatively damaged DNA lesion that has been associated with the development of aging, cancer and some degenerative diseases. Here, we present the first liquid chromatography-tandem mass spectrometry method that enables the simultaneous measurement of its repair products in plasma and saliva, namely 8-oxo-7,8-dihydroguanine (8-oxoGua) and 8-oxodGuo. Using this method, we investigated the underlying transport mechanism of the repair products of oxidatively damaged DNA between cellular compartments and biological matrices. Plasma, saliva and urine samples were collected concurrently from 57 healthy subjects. Various deproteinization methods were evaluated, and the precipitants acetonitrile and sodium hydroxide-methanol were, respectively, selected for plasma and saliva samples due to their effect on recovery efficiencies and chromatography. The mean baseline concentrations of 8-oxoGua and 8-oxodGuo in plasma were demonstrated to be 0.21 and 0.016 ng/mL, respectively, while in saliva they were 0.85 and 0.010 ng/mL, respectively. A relatively high concentration of 8-oxoGua was found in saliva with a concentration factor (CF, concentration ratio of saliva to plasma) of 4 as compared to that of 8-oxodGuo (CF: 0.6), implying that 8-oxoGua in plasma may be actively transported to saliva, whereas 8-oxodGuo was most dependent on a passive diffusion. Good correlations between urine and plasma concentrations were observed for 8-oxoGua and 8-oxodGuo, suggesting that blood was a suitable matrix in addition to urine. Significant correlation between 8-oxoGua and 8-oxodGuo in urine was only observed when the concentrations were not corrected for urinary creatinine, raising the issue of applicability of urinary creatinine to adjust 8-oxoGua concentrations.