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(0.625~10.000μg·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(10~50μg·mL^(-1))和金胺O(0.6~1.2μg·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2.50μg·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5μg·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。10μg·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0.625μg·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2.5μg·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,30~50μg·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;0.9~1.2μg·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,10~50μg·mL^(-1)金橙Ⅱ可导致L5178Y细胞微核率显著增加(P<0.05、0.01)。30~50μg·mL^(-1)金橙Ⅱ可导致S期的L5178Y细胞比例增加;金胺O自0.6μg·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 orangeⅡand auramine O using the fluctuation Ames test based on Salmonella typhimurium and the in vitro micronucleus test based on L5178 Y mouse lymphoma cells.Methods A series of concentrations of orangeⅡand auramine O(0.625—10.000μg·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,L5178 Y cells were treated with orangeⅡ(10—50μg·mL^(-1))and auramine O(0.6—1.2μg·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 OrangeⅡfrom 2.50μg·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.OrangeⅡsince 5μg·mL^(-1)could significantly increase the number of TA100 reverse mutation pores in-S9 treatment group(P<0.01).Auramine O of 10μg·mL^(-1)could significantly increase the number of TA98 reverse mutation pores in-S9 treatment group(P<0.01).Auramine O from 0.625μg·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 2.5μg·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—50μg·mL^(-1)orangeⅡcould significantly increase the micronucleus rate of L5178 Y cells(P<0.01).Auramine O of 0.9—1.2μg·mL^(-1)could significantly increase the micronucleus rate of L5178 Y cells(P<0.01).In the+S9 treatment group,orangeⅡof 10—50μg·mL^(-1)could significantly increase the micronucleus rate of L5178 Y cells(P<0.05,0.01).OrangeⅡof 30—50μg·mL^(-1)could increase the proportion of L5178 Y cells in S phase,Auramine O at 0.6μg·mL^(-1)could incre
作者
唐茵茹
王亚楠
王曼虹
黄芝瑛
汪祺
文海若
TANG Yinru;WANG Yanan;WANG Manhong;HUANG Zhiying;WANG Qi;WEN Hairuo(School of Pharmaceutical Sciences,Sun Yat-sen University,Guangzhou 510006,China;National Institutes for Food and Drug Control,Beijing 100050,China)
出处
《药物评价研究》
CAS
2022年第3期434-441,共8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)。
关键词
金橙Ⅱ
金胺O
染料
碱基突变
波动Ames试验
体外流式微核试验
orangeⅡ
auramine O
dyes
gene mutation
fluctuation Ames test
in vitro micronucleus assay by flow cytometry
