中草药对DNA氧化损伤水平的微分脉冲伏安法测定  

Determination of the Levels of DNA Damage Induced by Chinese Herbal Medicine Using Differential Pulse Voltammetry

作　　者：利健文[1] 陈任宏[1] 崔丽京[1] 汪小根[1] 

机构地区：[1]广东食品药品职业学院中药和生物系,广东广州510520

出　　处：《分析测试学报》2012年第6期675-679,共5页Journal of Instrumental Analysis

基　　金：广东省科技计划项目(2011B020314011)

摘　　要：采用微分脉冲伏安法(DPV)研究了中草药对脱氧核糖核酸分子(DNA)的损伤效应。在pH 5.0的磷酸盐缓冲液中,采用DPV法研究了8-羟基脱氧鸟苷(8-OHdG)在玻碳电极上的伏安行为,发现8-OHdG在+0.5 V电位处产生一灵敏的微分脉冲阳极氧化峰。该氧化峰的峰电流与8-OHdG的浓度在1.0×10-6～7.1×10-4mol/L范围内呈良好的线性关系,r=0.999 8,检出限(S/N=3)为3.5×10-7mol/L。将该方法应用于暴露于浓度均为40 g生药/L的甘草、金樱子、杜仲、半夏、马钱子提取液2 h的小牛胸腺DNA(ctDNA)中8-OHdG的分析,以及连续灌服低剂量和高剂量马钱子提取液30 d的昆明小鼠血中8-OHdG的分析。结果发现甘草、金樱子、杜仲、半夏提取液对ctDNA无氧化损伤作用,马钱子提取液能引起ctDNA氧化损伤形成8-OHdG,平均水平为(3.2±0.2)μmol/L。长期服用低剂量和高剂量马钱子提取液的小鼠,其血液中8-OHdG的平均水平分别为(2.0±0.1)、(5.3±0.3)μmol/L,表明马钱子具有潜在的遗传毒性。Effects of DNA damage induced by Chinese herbal medicine were studied by differential pulse voltammetric （DPV） method. The vohammeitric behaviour of 8-hydorxy-2＇-deoxyano-sine （ 8- OHdG） in the phosphate buffer（pH 5.0） was investigated at a glassy carbon electrode by cyclic vol- tammetry（CV） and DPV. A well-defined oxidation peak of 8-OHdG at ＋ 0. 5 V was found, and its current intensity was proportional to concentration of 8-OHdG in the range of 1.0 ×10 -6 - 7.1 × 10 -4 mol/L. The regression equation was Ip（μA） =0. 004 3c（mol/L） ＋4 × 10-8（r =0. 999 8） and the detection limit（ S/N = 3 ） was 3.5 × 10-7 mol/L. The method was applied in analysis of the levels of 8-OHdG in the calf thymus DNA（ctDNA） exposed to concentration of 40 g/L of glycyrrhiza, cherokee rose, eucommia ulmoides, pinellia, nux vomica extract for 2 h, respectively, and the blood of Kunming mice exposed to low and high concentration of nux vomica extract by mouth injection for 30 con- secutive days. The results showed that glycyrrhiza, cherokee rose, eucommia ulmoides, pinellia ex- tract could not cause ctDNA oxidative damage, and nux vomica extract can cause DNA oxidative damage to the formation of 8-OHdG in the average level of （ 3.2 ±0. 2 ） μmol/L. The average levels of 8- OHdG were （2. 0 ± 0. 1 ） μmol/L and （5.3± 0.3 ）μmol/L in the blood after a long-term administration of low concentration and high concentration of nux vomica extracts of Kunming mice, respectively. The study indicated that nux vomica contained the potential genotoxicity.

关 键 词：微分脉冲伏安法(DPV) 中草药 DNA氧化损伤 8-羟基脱氧鸟苷(8-OHdG) 遗传毒性 

分 类 号：O657.1[理学—分析化学] O629.74[理学—化学]