染料溶剂红207的体外致突变风险评价  

In vitro mutagenicity risk assessment of dye solvent red 207

作　　者：汪祺[1] 闫明 王亚楠 鄂蕊 胡燕平[1] 文海若 马双成[1] WANG Qi;YAN Ming;WANG Yanan;AO Rui;HU Yanping;WEN Hairuo;MA Shuangcheng(National Institutes for Food and Drug Control,Beijing 100050,China;China Pharmaceutical University,Nanjing,211198,China)

机构地区：[1]中国食品药品检定研究院,北京100050 [2]中国药科大学,江苏南京211198

出　　处：《药物评价研究》2020年第12期2416-2421,共6页Drug Evaluation Research

基　　金：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)。

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(0.625~10.000μg/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(2.5~20.0μg/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2.5~20μg/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。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(L5178 Y).Methods Different concentrations of solvent red 207(0.625—10.000μg/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 oC.In addition,solvent red 207(2.5—20.0μg/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 8 th day after treatement.Results Both in the absence and presence of S9 metabolic conditions,the solvent red 207 ranges between 0.625—10μg/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—20μg/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.

关 键 词：溶剂红207 致癌性 碱基突变 波动Ames试验 Pig-a基因突变试验 

分 类 号：R992[医药卫生—毒理学]