基于彗星实验的洛克沙胂对秀丽隐杆线虫胚胎细胞DNA损伤的研究

为评价洛克沙胂的遗传毒性,采用单细胞凝胶电泳(彗星实验),研究了不同浓度的洛克沙胂对秀丽隐杆线虫胚胎细胞脱氧核醣核酸(DNA)的损伤作用。提取秀丽隐杆线虫的胚胎细胞,分别暴露于0(空白对照)、50、250、500μg·L~(-1)含洛克沙胂的溶液染毒1 h。用彗尾DNA百分比含量(TDNA%)、彗星尾长(TL)和Olive尾矩(OTM)作为DNA损伤的指标。实验结果表明,与空白对照组比较,处理组中彗尾DNA百分比含量、彗星尾长以及Olive尾矩显著增加(P0.01)。随着洛克沙胂浓度的增加,彗尾DNA百分比含量、彗星尾长以及Olive尾矩逐渐增加,其相关系数r0.99,说明在实验浓度范围内,存在极显著的浓度-效应关系。洛克沙胂对秀丽隐杆线虫胚胎细胞DNA具有损伤作用,彗星实验操作简便、快速、灵敏度高,能够反映出洛克沙胂的遗传毒性。因此,通过彗星实验建立实验室检测洛克沙胂遗传毒性的方法具有可行性。

Comet Assay Study on DNA Damage of Embryonic Cells in Caenorhabditis elegans Induced by Roxarsone

To evaluate the genotoxicity of roxarsone, the present study investigated the damage effects of different concentrations of roxarsone on deoxyribonucleic acid (DNA) of embryonic cells in Caenorhabditiselegansby using the single cell gel￣electrophoresis (SCGE). The extracted embryonic cells of Caenorhabditis elegans were exposed to 0 (blank control), 50, 250, 500 μg?L￣1 roxarsone for 1 h, respectively. The percentage of comet tail DNA (TD￣NA%), the comet tail length (TL) and Olive tail moment (OTM) were used as the indicators of DNA damage. The results showed that, TDNA percentage, TL and OTM in all treatment groups increased significantly compared with these in the control group (P<0.01). TDNA percentage, TL and OTM increased gradually according to the in￣creased concentrations of roxarsone. The correlation coefficient of r greater than 0.99, which indicated that the con￣centration￣effect relationship was observed in the experimental concentration range. Roxarsone had damage effect on DNA of embryonic cells in Caenorhabditiselegans. Comet assay was simple, fast, and highly￣sensitive to reflect the genotoxicity of roxarsone. Thus, as a laboratory based method, it is practical to detect the genotoxicity of roxar￣sone by using comet assay.