[关键词]
[摘要]
目的 使用基于鼠伤寒沙门氏菌的Ames波动试验和小鼠淋巴瘤细胞(L5178Y)的体外Pig-a基因突变试验评价新型染料溶剂红207的碱基突变风险。方法 不同质量浓度的溶剂红207(0.625~10.000 μg/mL)分别与TA98和TA100混合于96孔培养板,在37℃条件下作用72 h后判断其对突变菌落数的影响。在优化后的体外L5178Y细胞Pig-a基因突变实验中,溶剂红207(2.5~20.0 μg/mL)分别与L5178Y细胞在有或无S9代谢活化条件下作用24 h或4 h,于给药后第8天评价其是否可诱导哺乳动物细胞Pig-a基因的突变频率增加。结果 无和有S9代谢活化条件下,溶剂红207可导致TA98和TA100回复性突变孔数显著增加(P<0.05、0.01、0.001),且存在浓度效应相关性,代谢活化后增加更为明显。在非S9代谢活化条件下,溶剂红207各浓度组Pig-a基因突变率与阴性对照组比较无显著性差异;在S9代谢活化条件下,溶剂红207质量浓度范围为2.5~20 μg/mL时,可导致Pig-a基因突变率显著增加(P<0.01、0.001),且存在浓度效应关系。结论 首次使用体外高通量试验方法评价溶剂红207的致突变风险,发现其存在体外致突变性且经I相代谢活化后致突变性进一步增强。
[Key word]
[Abstract]
Objective To evaluate the base-pair mutation risk of a novel medicinal dye solvent red 207 by fluctuation Ames test based on Salmonella typhimurium and the in vitro Pig-a gene mutation assay based on mouse lymphoma cells (L5178Y). Methods Different concentrations of solvent red 207 (0.625-10.000 μg/mL) were mixed with TA98 and TA100 in 96-well culture plates, and its effects to the number of revertants were determined after a 72 h incubation at 37 ℃. In addition, solvent red 207 (2.5-20.0 μg/mL) were treated with L5178Y cells under the conditions with or without metabolic activation for 24 h or 4 h, and its effects to the frequency of Pig-a gene mutation in mammalian cells were evaluated on the 8th day after treatement. Results Both in the absence and presence of S9 metabolic conditions, the solvent red 207 ranges between 0.625-10 μg/mL led to significant increase in the number of TA98 and TA100 revertants (P<0.05, 0.01 and 0.001), and there was a concentration-effect correlation, and the extent of increase in the number of revertants are more obvious after the metabolism. In absence of S9 activation, the Pig-a gene mutation rates in cells treated with different concentrations of solvent red 207 showed no difference comparing to the vehicle control group; In the presence of S9 activation, solvent red 207 ranges between 2.5-20 μg/mL induced increase of the frequency of Pig-a gene mutation, and there is a certain concentration-effect relationship (P<0.01 and 0.001). Conclusion This study for the first time used the in vitro high-throughput test methods to evaluate the mutagenic risk of solvent red 207, and suggested that the solvent red 207 is mutagenic, while the mutagenicity could be further enhanced after the phase I metabolism.
[中图分类号]
R992
[基金项目]
国家“重大新药创制”科技重大专项(2018ZX09201017);国家自然科学基金资助项目(81503347)