体内碱性彗星试验法检测纳米氧化锌遗传毒性

Detection of genotoxicity of nano zinc oxide by alkaline comet assay in vivo

目的利用体内碱性彗星试验方法,检测长期给予纳米氧化锌颗粒(ZnO NPs)对大鼠的遗传毒性作用。方法结合扩展一代生殖毒性试验,使用13周龄的亲代SD大鼠,经口每天灌胃给予ZnO NPs 0、7、50、350 mg/kg(纳米尺度分散状态最大浓度),记录体质量变化并观察。70 d后每个剂量组取亲代大鼠雌雄各10只处死后取乙二胺四乙酸抗凝全血,采用彗星试验试剂盒进行制片,用定量分析专业软件(Casp)进行彗星结果分析。结果与溶剂对照组比较,雄性大鼠高剂量组外周血DNA损伤细胞率和尾部DNA含量百分比明显上升,分别为28.60%和(36.38±5.84)%,差异有统计学意义(P<0.05);雌性大鼠高剂量组外周血DNA损伤细胞率和尾部DNA含量百分比明显上升,分别为27.31%和(18.80±2.96)%,差异有统计学意义(P<0.05)。结论在350 mg/kg剂量下ZnO NPs体内碱性彗星试验结果阳性。

Objective To detect the genotoxic effects of long-term administration of nano zinc oxide nanoparticles(ZnO NPs)particles on rats by using the alkaline comet assay in vivo.Methods In combination with the extended generation reproductive toxicity test,13-week-old parental SD rats were administered orally with ZnO NPs at 0,7,50,and 350 mg/kg(maximum concentration of the nanoscale dispersion),and body weight changes were recorded and observed.After 70 days,10 male rats and 10 female rats were taken from each dose group and sacrificed.Ethylene diamine tetraacetic acid anticoagulated whole blood was taken from each rat.The comet assay kit was used for tableting,and the result of comet analysis were analyzed by quantitative analysis software(Casp).Results Compared with the vehicle control group,peripheral blood DNA damage cell rate and tail DNA of the high-dose group of male rats increased significantly,which were 28.60%and(36.38±5.84)%,respectively(P<0.05).Peripheral blood DNA damage cell rate and tail DNA of the high-dose group of female rats increased significantly,which were 27.31%and(18.80±2.96)%,respectively(P<0.05).Conclusion The result of alkaline comet assay in ZnO NPs were positive at 350 mg/kg group.