U-SENS^(TM)试验方法的建立与探讨  

Establishment and Discussion of U-SENS^(TM) Test Method

作　　者：陈舒怀 段名扬 高行 匡荣 CHEN Shuhuai;DUAN Mingyang;GAO Hang;KUANG Rong(Zhejiang Institute for Food and Drug Control,NMPA Key Laboratory for Animal Alternative Testing Technology of Cosmetics,Hangzhou 310052,China)

机构地区：[1]浙江省食品药品检验研究院,国家药品监督管理局化妆品动物替代试验技术重点实验室,杭州310052

出　　处：《中国现代应用药学》2021年第24期3087-3090,共4页Chinese Journal of Modern Applied Pharmacy

基　　金：国家药品监督管理局化妆品动物替代试验技术重点实验室开放性课题(国浙药重[2020]7号)。

摘　　要：目的建立符合中国实际情况的U-SENS^(TM)试验方法,并验证新试验方法的可靠性。方法参考OECD指导原则442E附录II U-SENS^(TM)试验方法,根据其中的QC模板,建立以不同浓度松香酸(20,30,40,50,60,100μg·mL^(–1))为阳性剂的试验系统进行10次试验,应用流式细胞仪技术在人源性淋巴瘤细胞系U937探究最适阳性浓度,并对糖精、对苯二胺、肉桂醇等8个已知其致敏性的化合物进行验证。结果40,50μg·mL^(–1)松香酸能在较小的细胞毒性条件下,表现出稳定的阳性反应,并能对8个已知其致敏性的化合物能进行正确分类。结论以松香酸为阳性剂建立U-SENS^(TM)试验系统可行,可选用终浓度40μg·mL^(–1)或50μg·mL^(–1)作为阳性对照浓度,确保该试验系统能对受试物的皮肤致敏性进行准确分类。OBJECTIVE To establish the U-SENS^(TM)test method which is in line with the actual situation of China,and verify the reliability of the new test method.METHODS Refer to U-SENS^(TM)test method in OECD guideline 442E Appendix II,a test system with different concentrations of abietic acid(20,30,40,50,60,100μg·mL^(–1))as the positive agent was established for 10 tests according to the QC template,the optimum positive concentration of human lymphoma cell line U937 was explored by flow cytometry.And eight compounds with known sensitization,such as saccharin,p-phenylenediamine and cinnamyl alcohol were verified.RESULTS The 40,50μg·mL−1 abietic acid could show stable positive reaction under the condition with low cytotoxicity,and could correctly classify 8 compounds with known sensitization.CONCLUSION U-SENS^(TM)test system is established with abietic acid as positive agent,and 40 or 50μg·mL−1 can be chosen as the positive control concentration to ensure that the test system can accurately classify the skin sensitization of the test object.

关 键 词：U937 流式细胞术 化妆品替代 

分 类 号：R991[医药卫生—毒理学]