茜素型蒽醌化合物体外Pig-a基因突变性试验研究  

Mutagenic risk evaluation of alizarin-type anthraquinone using in vitro Pig-a mutation assay

作　　者：闫明 叶倩 王雪[1] 汪祺[1] 文海若 YAN Ming;YE Qian;WANG Xue;WANG Qi;WEN Hairuo(National Institutes for Food and Drug Control,Beijing 100050,China)

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《药物评价研究》2022年第7期1227-1232,共6页Drug Evaluation Research

基　　金：国家自然科学基金资助项目(81503347);国家“十三五”“重大新药创制”专项(2018ZX09201017)

摘　　要：目的对具有相同母核结构的7种茜素型蒽醌化合物开展体外Pig-a基因突变试验,分析不同茜素型蒽醌化合物取代基结构与其致突变性的关联。方法小鼠淋巴瘤细胞L5178Y(tk+/--3.7.2.C)分别与系列浓度的茜草素、异茜草素、甲基异茜草素、羟基茜草素、甲基异茜草素-1-甲醚、茜草素-1-甲醚和光泽汀作用4 h(有S9)或24 h(无S9),给药24 h后应用细胞计数仪进行计数,计算细胞相对倍增速率(RPD)评价受试物细胞毒性;细胞培养8 d后经APC-anti-CD45和PE-anti-CD90.2抗体孵育后,使用流式细胞仪检测细胞突变(CD45+CD90.2−)率。结果所有受试物在有或无代谢活化条件下所设浓度组RPD均大于50%,未见明显细胞毒性作用,可排除试验中假阳性结果。在无S9代谢活化条件下,光泽汀(16μg·mL^(−1))组、羟基茜草素(10μg·mL^(−1))组、甲基异茜草素-1-甲醚(31.25μg·mL^(−1))组Pig-a基因突变率与溶媒对照组比较显著升高(P<0.05、0.01、0.001);在有S9代谢活化条件下,光泽汀(4、8μg·mL^(−1))、羟基茜草素(10μg·mL^(−1))、茜草素-1-甲醚(3.75、15.00μg·mL^(−1))、甲基异茜草素(12.5、25.0、50.0、100.0μg·mL^(−1))、甲基异茜草素-1-甲醚(15、30、60μg·mL^(−1))、异茜草素(2.50、5.00μg·mL^(−1))组Pig-a基因突变率与溶媒对照组比较显著升高(P<0.05、0.01、0.001)。结论羟基取代基所在位点是茜素型蒽醌化合物致突变性强弱的决定性因素,其体内致突变性有待进一步确证。Objective Pig-a gene mutation asay was carried out on seven alizarin type anthraquinone compounds with the same parent nuclear structure in vitro and the relationship between the substituent structure and the mutagenicity were analysed.Methods Alizarin,xanthopurpurin,rubiadin,purpurin,rubiadin-1-methyl,alizarin-1-methyl and lucidin were treated on mouse lymphoma cell L5178Y(tk+/--3.7.2.C)for 4 hours(-S9)or 24 hours(+S9).Cell counting plate was used to count 24 h after administration,and the relative cell multiplication rate(RPD)was calculated to evaluate the cytotoxicity of the tested samples.Cells were cultured for 8 days and APC anti-CD45 antibody/PE-anti-CD90.2 antibody were used to detected mutant cells(CD45+CD90-)by flow cytometry.Results The RPD of all tested substances in the concentration group with or without metabolic activation was greater than 50%,and no obvious cytotoxicity was observed,so false positive results in the test could be excluded.In the absence of S9,the mutant cells(CD45+CD90.2−)were significantly increased by lucidin(16μg·mL^(−1)),purpurin(10μg·mL^(−1))and rubiadin-1-methyl ether(31.25μg·mL^(−1))compared with solvent control group.In the presence of S9 metabolic activation,the mutation rate of Pig-a gene in lubutin(4,8μg·mL^(−1)),hydroxyl alizarin(10μg·mL^(−1)),alizarin-1-methyl ether(3.75,15.00μg·mL^(−1)),methylisoalizarin(12.5,25.0,50.0,100.0μg·mL^(−1)),methylisomadicin-1-methyl ether(15,30,60μg·mL^(−1))and isomadicin(2.50,5.00μg·mL^(−1))groups was significantly higher than that in solvent control group(P<0.05,0.01,0.001).Conclusion The location of hydroxyl substituents is a decisive factor in themutagenicity of alizarin type anthraquinone compounds,while their in vivomutagenicity need to be further investigated.

关 键 词：茜素型蒽醌 致突变性 L5178Y细胞 体外Pig-a基因突变试验 茜草素 异茜草素 甲基异茜草素 羟基茜草素 甲基异茜草素-1-甲醚 茜草素-1-甲醚 光泽汀 

分 类 号：R994[医药卫生—毒理学]