目的 使用基于鼠傷寒沙門氏菌的Ames波動試驗和小鼠淋巴瘤細胞（L5178Y）的體外微核試驗評價2種染料金橙Ⅱ和金胺O的遺傳毒性風險。方法 在-/+S9處理下，不同質(zhì)量濃度的金橙Ⅱ和金胺O（0.625~10.000 μg·mL^-1）分別與鼠傷寒沙門氏菌組氨酸營養(yǎng)缺陷型菌株TA98和TA100混合后接種于96孔板中，37℃下孵育72 h后判斷其對細菌回復突變的影響；在體外微核試驗中，在-/+S9處理下，金橙Ⅱ（10~50 μg·mL^-1）和金胺O（0.6~1.2 μg·mL^-1）分別與L5178Y細胞作用4 h或24 h，并在給藥后24 h收集細胞進行流式細胞術檢測。結(jié)果 金橙II自2.50 μg·mL^-1起可引起-S9和+S9處理組的TA98回復性突變孔數(shù)顯著增加（P<0.05、0.01），代謝活化后增加幅度更為明顯；自5 μg·mL^-1起可引起-S9處理組的TA100回復性突變孔數(shù)顯著性增加（P<0.01），作用均呈濃度相關性。10 μg·mL^-1金胺O可引起-S9處理組的TA98回復性突變孔數(shù)顯著增加（P<0.01）；自0.625 μg·mL^-1起即可引起+S9處理組的TA98回復性突變孔數(shù)顯著增加（P<0.05、0.01），且存在濃度相關性；自2.5 μg·mL^-1起可引起+S9處理組的TA100回復性突變孔數(shù)顯著性增加（P<0.05、0.01），且存在濃度相關性。在-S9處理組中，30~50 μg·mL^-1金橙Ⅱ可引起L5178Y細胞微核率顯著增加（P<0.01），且存在濃度相關性； 0.9~1.2 μg·mL^-1金胺O可引起L5178Y細胞微核率顯著增加（P<0.01），且存在濃度相關性。在+S9處理組中，10~50 μg·mL^-1金橙Ⅱ可導致L5178Y細胞微核率顯著增加（P<0.05、0.01）。30~50 μg·mL^-1金橙Ⅱ可導致S期的L5178Y細胞比例增加；金胺O自0.6 μg·mL^-1起即可導致L5178Y細胞亞二倍體細胞核率增加。結(jié)論 金橙Ⅱ和金胺O均存在一定的遺傳毒性風險。

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

國家“重大新藥創(chuàng)制”科技重大專項（2018ZX09201017）；國家自然科學基金資助項目（81503347）