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）的体外微核试验评价2种染料金橙Ⅱ和金胺O的遗传毒性风险。方法 在-/+S9处理下，不同质量浓度的金橙Ⅱ和金胺O（0.625~10.000 μg·mL^-1）分别与鼠伤寒沙门氏菌组氨酸营养缺陷型菌株TA98和TA100混合后接种于96孔板中，37℃下孵育72 h后判断其对细菌回复突变的影响；在体外微核试验中，在-/+S9处理下，金橙Ⅱ（10~50 μg·mL^-1）和金胺O（0.6~1.2 μg·mL^-1）分别与L5178Y细胞作用4 h或24 h，并在给药后24 h收集细胞进行流式细胞术检测。结果 金橙II自2.50 μg·mL^-1起可引起-S9和+S9处理组的TA98回复性突变孔数显著增加（P<0.05、0.01），代谢活化后增加幅度更为明显；自5 μg·mL^-1起可引起-S9处理组的TA100回复性突变孔数显著性增加（P<0.01），作用均呈浓度相关性。10 μg·mL^-1金胺O可引起-S9处理组的TA98回复性突变孔数显著增加（P<0.01）；自0.625 μg·mL^-1起即可引起+S9处理组的TA98回复性突变孔数显著增加（P<0.05、0.01），且存在浓度相关性；自2.5 μg·mL^-1起可引起+S9处理组的TA100回复性突变孔数显著性增加（P<0.05、0.01），且存在浓度相关性。在-S9处理组中，30~50 μg·mL^-1金橙Ⅱ可引起L5178Y细胞微核率显著增加（P<0.01），且存在浓度相关性； 0.9~1.2 μg·mL^-1金胺O可引起L5178Y细胞微核率显著增加（P<0.01），且存在浓度相关性。在+S9处理组中，10~50 μg·mL^-1金橙Ⅱ可导致L5178Y细胞微核率显著增加（P<0.05、0.01）。30~50 μg·mL^-1金橙Ⅱ可导致S期的L5178Y细胞比例增加；金胺O自0.6 μg·mL^-1起即可导致L5178Y细胞亚二倍体细胞核率增加。结论 金橙Ⅱ和金胺O均存在一定的遗传毒性风险。

Objective To evaluate the genotoxicity of two dyes orange II and auramine O using the fluctuation Ames test based on Salmonella typhimurium and the in vitro micronucleus test based on L5178Y mouse lymphoma cells. Methods A series of concentrations of orange II and auramine O (0.625-10.000 μg·mL^-1) were mixed with TA98 and TA100, respectively, and then cultured into 96-well plates. After incubating at 37℃ for 72 h, their effects on mutagenic respond were determined. In addition, L5178Y cells were treated with orange II (10-50 μg·mL^-1) and auramine O (0.6-1.2 μg·mL^-1) for 4 h or 24 h without or with metabolic activation, and cells were collected for in vitro micronucleus assay by flow cytometry. Results Orange II from 2.50 μg·mL^-1 could significantly increase the number of TA98 reverse mutation pores in - S9 and + S9 treatment groups (P < 0.05, 0.01), and the increase was more obvious after metabolic activation. Orange II since 5 μg·mL^-1 could significantly increase the number of TA100 reverse mutation pores in -S9 treatment group (P < 0.01). Auramine O of 10 μg·mL^-1 could significantly increase the number of TA98 reverse mutation pores in -S9 treatment group (P < 0.01). Auramine O from 0.625 μg·mL^-1 could significantly increase the number of TA98 reverse mutation pores in +S9 treatment group (P < 0.05, 0.01), and there was a concentration correlation. Auramine O from 2.5 μg·mL^-1 could significantly increase the number of TA100 reverse mutation pores in +S9 treatment group (P < 0.05, 0.01), and there was a concentration correlation. In the - S9 treatment group, 30-50 μg·mL^-1 orange Ⅱ could significantly increase the micronucleus rate of L5178Y cells (P < 0.01). Auramine O of 0.9-1.2 μg·mL^-1 could significantly increase the micronucleus rate of L5178Y cells (P < 0.01). In the +S9 treatment group, orange Ⅱ of 10-50 μg·mL^-1 could significantly increase the micronucleus rate of L5178Y cells (P < 0.05, 0.01). Orange Ⅱ of 30-50 μg·mL^-1 could increase the proportion of L5178Y cells in S phase, Auramine O at 0.6 μg·mL^-1 could increase the rate of sub diploid nucleus of L5178Y cells. Conclusion Orange II and auramine O have genotoxic potentials.

R965.2

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）