【摘要】 目的:应用蛋白组学技术筛选用于评价化合物皮肤急性刺激性的特异性生物标志物。方法:选取2种欧盟列举的具有代表性的皮肤刺激性化合物十二烷基硫酸钠(SLS)、10-十一碳烯酸(UA)和1种非刺激性化合物3,3’-二硫代二丙酸(DA),严格按照体外评价皮肤刺激性替代方法的检测流程,使人体皮肤组织暴露于化合物15 min,然后裂解组织,提取蛋白,进行二维电泳;选择蛋白表达差异点,进行质谱鉴定,并用real-time RT-PCR和Western blotting验证差异蛋白的表达。结果:在鉴定的8个蛋白点中,SLS和UA刺激使人体皮肤热休克蛋白27(HSP27)的表达显著上调,与Western blotting结果一致。结论:HSP27有可能成为评价皮肤刺激性体外替代方法体系的候选生物标志物,其表达强度与刺激强度的关系需进一步探讨。更多还原

【Abstract】 AIM: To discover the novel endpoints for skin irritation responses by two-dimensional electrophoresis( 2DE)-based proteomics. METHODS: 2DE-based proteomics was used to analyze the protein expression in human skin exposed to sodium lauryl sulfate( SLS),10-undecenoic acid( UA) and 3,3’-dithiodipropionic acid( DA) following the test protocol of the European Centre for the Validation of Alternative Methods( ECVAM). The altered protein expression was validated by real-time RT-PCR and Western blotting. RESULTS: Heat-shock protein 27( HSP27) was up-regulated most significantly among the 8 identified proteins,consistent with the results of Western blotting. CONCLUSION: HSP27 may represent a potential marker or additional endpoint for the hazard assessment of skin irritation caused by chemical products.更多还原