毒理学杂志

目的 使用BALB/c小鼠建立皮肤致敏性局部淋巴结试验(LLNA:DA)方法，为皮肤致敏性替代方法在国内的发展和应用积累数据。方法 使用8～9周龄雌性BALB/c小鼠，每组4只，测试9个受试物(己基肉桂醛、2,4-二硝基氯苯、间苯二甲酸二甲酯、丁子香酚、反式肉桂醛、甲醛、二氯化钴、重铬酸钾和异丙醇)的皮肤致敏性，每个受试物设3个浓度。染毒结束后分离每只小鼠双耳耳廓、双侧颌下淋巴结并称重，测定淋巴结ATP发光量，进行受试物致敏性判断及致敏性分级，并与替代方法验证机构间协调委员会(ICCVAM)致敏分级数据进行比较。结果 25%己基肉桂醛、0.3%2,4-二硝基氯苯、25%反式肉桂醛、13.5%甲醛、5%二氯化钴、0.3%和1%重铬酸钾耳廓重和淋巴结重增加差异有统计学意义(F_耳廓重=3.516等，F_淋巴结重=28.647等，P<0.05);4.05%甲醛、3%二氯化钴仅淋巴结重增加差异有统计学意义(F=47.453,F=31.108,P<0.05),耳廓重未见明显变化；提示这些浓度的受试物具有刺激性，且指标淋巴结重较耳廓重更灵敏。其他浓度的受试物未引起BALB/c小鼠淋巴结重和耳廓重有统计学意义的变化。己基肉桂醛、2,4-二硝基氯苯、丁子香酚、反式肉桂醛、甲醛、二氯化钴、重铬酸钾对BALB/c小鼠具有皮肤致敏性，而异丙醇、间苯二甲酸二甲酯对BALB/c小鼠无皮肤致敏性。与ICCVAM数据进行比较，致敏性结果灵敏度100%、特异度100%、准确度100%。结论 BALB/c小鼠LLNA:DA方法可以用于评价皮肤致敏性，鉴于动物品系与ICCVAM使用的CBA小鼠不同，本研究为使用BALB/c小鼠的皮肤致敏性局部淋巴结试验方法在国内的发展和应用积累了实验数据。

Objective To test and verify the local lymph node assay in BALB/c mice for skin sensitization evaluation. Methods Female BALB/c mice were organized into several groups of four mice per group. Nine test substance(three different concentration groups per chemical) were evaluated. The result were compared with the result of the international authoritative organization(ICCVAM). Results Ear punch weights and the bilateral draining auricular lymph nodes weights from mice treated with 25% hexyl cinnamaldehyde, 0.3% 2,4-dinitrochlororobenzene, 25% trans-cinnamaldehyde, 13.5% formaldehyde, 5% cobalt dichloride, 0.3% potassium dichromate, 1% potassium dichromate were increased significantly(F_{ear punch}=3.516, etc, F_{lymph nodes}=28.647, etc, P<0.05). The bilateral draining auricular lymph nodes weights from mice treated with 4.05% formaldehyde, 3% cobalt dichloride were increased significantly(F=47.453,F=31.108,P<0.05). Ear punch weights were not. For LLNA:DA in BALB/c mice, 7 chemicals(hexyl cinnamaldehyde, 2,4-dinitrochlororobenzene, eugenol, trans-cinnamaldehyde, formaldehyde, cobalt dichloride, potassium dichromate) were positive sensitization, and the others were negative. Compared with the result of ICCVAM, the accuracy of sensitization prediction was 100%. Conclusion LLNA:DA in BALB/c mice could be useful for evaluating sensitization potential. And more experimental data are accumulated to develop this assay in China.