小鼠局部淋巴结试验(BrdU-ELISA)改良法在2,4-二硝基氯代苯皮肤致敏性检测中的应用  

Application of modified local lymph node assay (BrdU-ELISA) in the detection of DNCB skin sensitization in mice

作　　者：秦珩 程洁[2] 靳苏香[1] 钱雯 胡艳辉 张成香 章婉 严婷 董昊 张璐璐[1,2] 张静姝 王玉邦 环飞 QIN Heng;CHENG Jie;JIN Su-xiang;QIAN Wen;HU Yan-hui;ZHANG Cheng-xiang;ZHANG Wan;YAN Ting;DONG Hao;ZHANG Lu-lu;ZHANG Jing-shu;WANG Yu-bang;HUAN Fei(The Safety Assessment and Research Center for Drugs of Jiangsu Province,Nanjing Jiangsu 211166,China;School of Public Health Nanjing Medical University,Nanjing Jiangsu 211166,China)

机构地区：[1]江苏省医药农药兽药安全性评价与研究中心,江苏南京211166 [2]南京医科大学公共卫生学院,江苏南京211166

出　　处：《毒理学杂志》2020年第2期138-142,共5页Journal of Toxicology

基　　金：江苏高校优势学科建设工程资助项目(苏政办发〔2014〕37号)。

摘　　要：目的采用小鼠局部淋巴结试验BrdU-ELISA改良法检测2,4-二硝基氯代苯(DNCB)的致敏性。方法 30只雌性BALB/c小鼠进行试验,DNCB设为1.00%、0.50%、0.25%和0.05%的浓度,同时设阴性对照组(AOO)及阳性对照组(25%已基肉桂醛),连续双耳背面染毒3 d,末次染毒后48 h采集耳后淋巴结称重并体外培养,同时体外给予BrdU溶液培养24 h,然后采用BrdU-ELISA试验方法检测淋巴增殖情况。结果与对照组比较,小鼠各剂量组各观察点的体重差异均无统计学意义(P>0.05);阴性对照组和阳性对照组的耳厚度增加不超过25%、阳性对照组的耳重相对于阴性对照组的耳重增加不超过25%;阴性对照组的刺激指数(SI1和SI2)<1.6,阳性对照组(25%已基肉桂醛)的刺激指数(SI1和SI2)>1.6,试验系统可靠;1%DNCB剂量组的耳厚度增加不超过25%,但相对于阴性对照组耳重增加超过25%,1%DNCB的刺激指数(SI1和SI2)>1.6;0.25%、0.50%DNCB剂量组的耳厚度增加不超过25%,且耳重相对于阴性对照组耳重增加不超过25%,0.25、0.50%DNCB的刺激指数(SI1和SI2)>1.6;0.05%DNCB剂量组的耳厚度增加不超过25%,且耳重相对于阴性对照组耳重增加不超过25%,0.05%DNCB的刺激指数(SI1和SI2)<1.6;DNCB的EC1.6=0.1738(SI2等于1.6时受试物的浓度)。结论 0.05%DNCB剂量组显示无刺激性与致敏性;0.25%、0.50%DNCB剂量组显示仅有致敏性,无刺激性;1.00%DNCB剂量组显示不仅有致敏性还有刺激性;DNCB的EC1.6在0.1~1之间,DNCB为强致敏物。Objective To detect DNCB skin sensitization with modified local lymph node assay(BrdU-ELISA) in mice. Methods Thirty female BALB/c mice were randomly divided into six groups: the negative control group(AOO) DNCB group(1.00%,0.50%,0.25,0.00%)and the positive control group(25% basal cinnamaldehyde). Test sample were applied on the dorsum of the bilateral mice ears for three consecutive days;after 48 hours, the lymph nodes of the mice were executed and weighted, then made into cell suspension, the lymph cell were cultured by BrdU solution in vitro, and the lymph cell proliferation was measured with BrdU-ELISA method after 24 h. Results Compared with the control group, the body weights for female mice for all treated groups are not significantly different. Compared with the negative control group, the ear thickness of the negative and positive control groups were increased by no more than 25%, and the ear weight of the negative and positive control groups were increased by no more than 25%. The stimulus indexes of negative control groups(SI1 and SI2) were less than 1.6, and the stimulus indexes of positive control group(SI1 and SI2, 25% hexyl cinnamaldehyde) were greater than 1.6, and the test system was reliable. Compared with the negative control group, the ear thickness of the 1% DNCB group was increased by no more than 25%, but the ear weight of the 1% DNCB group was increased by more than 25%. The stimulation indexes of 1% DNCB groups(SI1 and SI2) were greater than 1.6. The ear thickness of the 0.25% and 0.50% DNCB groups were increased by no more than 25%, and the ear weight of the 0.25% and 0.50% DNCB group were increased by no more than 25% compared with the negative control group. The stimulation indexes of 0.25% and 0.50% DNCB groups(SI1 and SI2) were greater than 1.6. Compared with the negative control group, the ear thickness of the 0.05% DNCB group was increased by no more than 25%, and the ear weight of the 0.05% DNCB dose group was increased by no more than 25%. The stimulation indexes of 0.05% DNCB gr

关 键 词：BALB/c小鼠 BRDU 致敏性 

分 类 号：R33[医药卫生—人体生理学]