溴脱氧尿嘧啶核苷掺入法作为皮肤致敏试验替代方法研究  

Study on using bromodeoxyuridine incorporation method as an alternative approach of skin sensitization test

作　　者：何国群[1] 杨杏芬[1] 郑穗生[1] 黄俊明[1] 杨颖[1] 谢晓平[1] 古梅英[1] 谭小华[1] 李庆[1] 

机构地区：[1]广东省疾病预防控制中心卫生毒理所,广东广州510300

出　　处：《中国职业医学》2011年第5期377-380,共4页China Occupational Medicine

基　　金：卫生公益性行业科研专项(200802080);广东省医学科学技术基金项目(A2006067)

摘　　要：目的探讨应用溴脱氧尿嘧啶核苷(BrdU)示踪和酶联免疫吸附测定(ELISA)的局部淋巴结试验(LLNA:Br-dU-ELISA法)替代LLNA(放射法)作为皮肤变态反应试验替代方法的可行性。方法雌性BALB/c小鼠随机分组,每组4只。选择27种受试物进行测试,每种受试物视情况设3～4个剂量组和1个溶剂对照组。将受试物连续3 d涂抹于小鼠双耳背,第5天腹腔注射BrdU标记液,第6天摘取小鼠耳部淋巴结,制备淋巴细胞悬液,采用ELISA技术测定BrdU掺入细胞的水平。选择3种受试物进行3次重复性试验以了解结果的重现性;将27种受试物的LLNA:BrdU-ELISA法与文献报告的LLNA(放射法)结果比较,分析方法的致敏性判别能力及分级结果的一致性和相关性。结果 LLNA:Br-dU-ELISA法结果重现性较好(F=2.890,P>0.05);与LLNA(放射法)相比,该方法对27种化学物的致敏性判别能力较好(灵敏度=95.00%,特异度=85.71%,符合率=92.59%),致敏性分级结果有较好的一致性和相关性(Kappa=0.575,Gamma=0.959)。结论 LLNA:BrdU-ELISA法可作为皮肤变态反应试验的替代方法应用于化学物皮肤致敏性评价。Objective To study the possibility of the local lymph node assay based on bromodeoxyuridine-ELISA （ LLNA ： BrdU-ELISA ） as an alternative method of skin sensitization test to replace the original LLNA （ radioisotope method）. Methods Female BALB/c mice were randomly selected （ n = 4 per concentration group）. 27 chemicals were tested, 3 or 4 concentration groups and 1 vehicle-treated control group were designed for each chemical. The mice were treated with the test substance at the dorsum of both ears daily for 3 consecutive days. At the fifth day the mice were injected intraperitoneally with bromodeoxyuridine （BrdU）. At the sixth day the draining auricular nodes were excised. Single lymphocyte suspension of lymph node cells were pre- pared, and analyzed with enzyme-linked immunosorbent assay （ELISA）to measure the levels of BrdU incorporation. 3 chemicals were repeatedly tested for 3 times to analyze the reproducibility of the LLNA ： BrdU-ELISA method. Compared with the LLNA （radioisotope method）, sensitization discriminative ability, consistency and correlation of sensitization classification results of LLNA ： BrdU-ELISA were analyzed. Results It was shown that the experimental results had a good repeatability （ F = 2. 890, P 〉 0. 05 ）. The test results showed that LLNA ： BrdU-ELISA had better ability to discriminate sensitization of 27 chemicals （ sensiti- vity =95.00% , specificity =85.71%, coincidence rate =92. 59% ）and had better consistency and correlation （Kappa =0. 575, Gamma = O. 959） compared with the LLNA （ radioisotope method）. Conclusion The results suggested that LLNA： BrdU-ELISA may be used as an alternative method for skin sensitization test to evaluate the contact-sensitizating potential of chemicals.

关 键 词：皮肤变态反应试验 替代方法 局部淋巴结试验 溴脱氧尿嘧啶核苷 

分 类 号：R758.2[医药卫生—皮肤病学与性病学]