【摘要】 目的研究苯并(a)芘(BaP)诱导大鼠产生(7R,8S)-二羟基-(9S,10R)-环氧-7,8,9,10-四氢苯并(a)芘(BPDE)-DNA加合物的生成。建立以血液为样品的检测染毒大鼠BPDE-DNA加合物的高效液相色谱法。方法选用清洁级SD大鼠,一次性腹腔注射苯并(a)芘二甲基亚砜溶液(以1%羧甲基纤维素钠为助溶剂)100 mg/kg,5 h后取股静脉血。提取抗凝全血中的DNA,采用琼脂糖凝胶电泳确认DNA的提取效果。将提取的DNA在0.1 nmol/L HCl、90℃恒温水浴箱中酸水解4 h,乙酸乙酯提取酸水解产物——四醇-苯并(a)芘,高效液相色谱法检测,最后用高效液相色谱-质谱法确认。结果BaP染毒组与溶剂对照组和阴性对照组比较,高效液相色谱检测有新的色谱峰产生,质谱确定其相对分子质量与四醇-苯并(a)芘一致。结论本实验的染毒方法能使大鼠产生BPDE-DNA加合物;建立的高效液相色谱法可以血液为样品检测BPDE-DNA加合物。更多还原

【Abstract】 Objective To study the benzo(a) pyrene(BaP) induced DNA adduct in rat and establish a method to measure the DNA adduct in blood by high performance liquid chromatography(HPLC).Methods The SD rats were treated with 100 mg/kg of BaP-DMSO(cosolvent:1% sodium carboxymethyl cellulose) once by i.p.,and the blood of femoral vein was collected 5 hours later.The blood DNA was extracted by kit and confirmed by agarose gel electrophoresis.After extraction,the DNA adducts were hydrolyzed in 0.1 nmol/L HCl at 90 ℃ for 4 hours.The acid-hydrolysis products(BP-tetrols) of DNA adducts were extracted by ethyl acetate and measured by HPLC,and finally confirmed by HPLC-MS.Results In chromatogram there were new peaks to occur for rats treated by BaP,with compared to control.By HPLC-MS,one of the new peaks was confirmed to be BP-tetrol.Conclusion The rats ingesting BaP in the way described in this experiment can form DNA adducts,and the adducts in blood can be detected by HPLC.更多还原