局部淋巴结试验(LLNA:BrdU-ELISA)对14种化学物刺激和致敏性的评价  

Local lymph node assay(LLNA:BrdU-ELISA)for evaluating the irritation and sensitization of 14 chemicals

作　　者：陈宁 黄泽愉 孙晓 谢天柱 陈润桦 郭煜堂 李俊鹏 冼静雯 吴熙 秦美蓉 王平 王晓炜 鲁艺 CHEN Ning;HUANG Ze-yu;SUN Xiao;XIE Tian-zhu;CHEN Run-hua;GUO Yu-tang;LI Jun-peng;XIAN Jing-wen;WU Xi;QIN Mei-rong;WANG Ping;WANG Xiao-wei;LU Yi(Shenzhen Institute for Drug Control,NMPA Key Laboratory for Monitoring and Evaluation of Cosmetics,GDMPA Key Laboratory of Animal Alternative Testing Technology on Cosmetics,Shenzhen Guangdong 518057,China)

机构地区：[1]深圳市药品检验研究院国家药品监督管理局化妆品监测评价重点实验室广东省药品监督管理局化妆品动物替代试验技术重点实验室,广东深圳518057

出　　处：《毒理学杂志》2022年第2期157-162,共6页Journal of Toxicology

基　　金：深圳市科技创委科技计划项目-基础研究(自由探索)(JCYJ20180302150138733);广东省药品监督管理局科技引导扶持项目(2018YDB32);广东省药品监督管理局重点实验室项目(2018ZDB13);深圳市药品检验研究院重点学科团队建设项目。

摘　　要：目的探讨BALB/c小鼠局部淋巴结试验LLNA:BrdU-ELISA改良法在化学物/化妆品皮肤刺激性和致敏性评价中的应用。方法采用BALB/c小鼠LLNA:BrdU-ELISA改良法,对14种化学物进行评价,并将结果与国际权威机构ICCVAM结果对照分析。结果阳性对照组(25%已基肉桂醛)的耳廓重和溶剂对照组相比增加不超过25%,刺激指数SI>1.6,试验成立;刺激性试验结果显示:1%DNCB剂量组和50%已基肉桂醛组相对于溶剂对照组耳重增加超过25%,其余化学物各剂量组耳重相对于溶剂对照增加未超过25%。致敏性评估结果显示:已基肉桂醛(10%,25%,50%),2,4-二硝基氯苯(0.1、0.3和1.0%),丁子香酚(10%、25%和50%),3-氨基苯酚(1%、3%和10%),异丁香酚(5%、10%和25%),甲醛(2.7%、8.1%和27%),戊二醛(0.2%、0.6%和2.0%),二氯化钴(1%、3%和5%),硫酸镍(1%、3%和10%),重铬酸钾(0.1%、0.3%和1,0%),反式肉桂醛(1%、3%和10%)11种化学物为致敏阳性,异丙醇、间苯二甲酸二甲酯、水杨酸甲酯为致敏阴性。致敏性预测准确率为100%(14/14),阳性致敏物预测率为100%(11/11),阴性致敏物预测率为100%(3/3),致敏分级准确率为86%(12/14)。结论BALB/c小鼠LLNA:BrdU-ELISA改良法可较好地评估化学物的致敏性,用于未来化学物致敏性检测。Objective To explore the application of BALB/c mice in local lymph node assay BrdU-ELISA modified method for the evaluation of skin sensitization and irritation of chemical and cosmetics.Methods Fourteen chemicals were evaluated using BALB/c mouse LLNA:BrdU-ELISA modified method,and the result were compared with ICCVAM result.Results Compare with the solvent control group,the ear weight of the positive control group(25%hexyl cinnamaldehyde)increased by no more than 25%,and the stimulation index SI of positive control group was greater than 1.6,the test system was reliable.Stimulation test result showed that ear weight increased by more than 25%in the 1%DNCB dose group and 50%hexyl cinnamaldehyde group compared to the solvent control group,while ear weight increased by less than 25%in the other chemical dose groups compared to the solvent control group.The results of sensitization of 11 chemics showed:2,4-dinitrochlorobenzene(0.1,0.3,1.0%),hexyl cinnamaldehyde(10%,25%,50%),eugenol(10%,25%,50%),3-aminophenol(1%,3%,10%),isoeugenol(5%,10%,25%),formaldehyde(2.7%,8.1%,27%),glutaraldehyde(0.2%,0.6%,2.0%),cobalt dichloride(1%,3%,5%),nickel sulfate(1%,3%,10%),potassium dichromate(0.1%,0.3%,1,0%),trans-cinnamaldehyde(1%,3%,10%)were positive,and the rest were negative.The sensitization prediction accuracy rate was 100%(14/14),the positive sensitizer prediction rate was 100%(11/11),the negative sensitizer prediction rate was 100%(3/3),and the sensitization grading accuracy rate is 86%(12/14).Conclusion This validation study indicates that BALB/c mouse could be used alternatively in the LLNA:BrdU-ELISA for the prediction of the skin sensitizing potential of chemicals.

关 键 词：局部淋巴结试验 皮肤致敏性 5-溴脱氧尿嘧啶核苷 酶联免疫测定 

分 类 号：R758.22[医药卫生—皮肤病学与性病学]