Fletcher S T, Baker V A, Fentem J H, Basketter D A, Kelsell D P
SEAC Toxicology Unit, Unilever Research, Sharnbrook, Bedfordshire MK44 1LQ, UK.
Toxicol In Vitro. 2001 Aug-Oct;15(4-5):393-8. doi: 10.1016/s0887-2333(01)00042-x.
There is a need to investigate the mechanistic basis of the human skin irritation response if relevant in vitro test systems for the predictive identification of skin irritation hazards are to be developed. Recent progress in genomics technologies mean that tools for the identification and investigation of important biochemical events in the processes of skin irritation are now available. The aim of this work was to identify genes (for further mechanistic investigation) which may be regulated in response to skin irritation, following exposure of the EpiDerm skin model to the known skin irritant sodium lauryl sulphate (SLS). EpiDerm cultures were treated in triplicate with a non-cytotoxic dose of SLS (0.1 mg/ml, as determined by the MTT assay and histological examination) for 15 min, 30 min, 1 h, 2 h, 3 h, 4 h and 24 h. Total RNA was extracted from the pooled EpiDerm cultures and used to probe Atlas human arrays (Clontech) covering approximately 3600 genes. Preliminary data indicated an up-regulation at early time points (15-30 min) of a number of genes involved in transportation (e.g. the sodium and chloride dependent taurine transporter) and receptors (e.g. ZAP70 and protocadherin 42 precursor). The gene encoding the UV excision repair protein and other DNA repair genes (e.g. DNA-directed RNA polymerase II) were up-regulated after 1-3 h, along with TGF beta 3 and other tumour suppressors, which play a role in cellular development and wound healing. At the later time points of 4-24 h, genes involved in protein translation (e.g. Cathepsin D receptor) and metabolism (e.g. CYP27A) were up-regulated. In addition, a number of genes were down-regulated in response to treatment with SLS, although these followed less of a time dependent pattern. These results indicate the differential regulation of a number of genes in response to treatment with SLS, some of which may provide additional clues to the molecular events underpinning the irritation response to this particular surfactant and possibly to other chemical irritants.
如果要开发用于预测性识别皮肤刺激危害的相关体外测试系统，就有必要研究人体皮肤刺激反应的机制基础。基因组学技术的最新进展意味着，现在已有工具可用于识别和研究皮肤刺激过程中的重要生化事件。这项工作的目的是在将EpiDerm皮肤模型暴露于已知的皮肤刺激物月桂醇硫酸酯钠（SLS）后，识别可能因皮肤刺激而被调控的基因（以便进一步进行机制研究）。用无细胞毒性剂量的SLS（0.1 mg/ml，通过MTT试验和组织学检查确定）对EpiDerm培养物进行一式三份处理，处理时间分别为15分钟、30分钟、1小时、2小时、3小时、4小时和24小时。从合并的EpiDerm培养物中提取总RNA，并用于探测覆盖约3600个基因的Atlas人类阵列（Clontech公司）。初步数据表明，在早期时间点（15 - 30分钟），一些参与转运的基因（如钠和氯依赖性牛磺酸转运体）和受体（如ZAP70和原钙黏蛋白42前体）出现上调。编码紫外线切除修复蛋白的基因和其他DNA修复基因（如DNA指导的RNA聚合酶II）在1 - 3小时后上调，同时TGFβ3和其他肿瘤抑制因子也上调，它们在细胞发育和伤口愈合中起作用。在4 - 24小时的较晚时间点，参与蛋白质翻译的基因（如组织蛋白酶D受体）和代谢基因（如CYP27A）上调。此外，一些基因在SLS处理后出现下调，尽管这些下调较少呈现时间依赖性模式。这些结果表明，许多基因在SLS处理后存在差异调控，其中一些基因可能为支撑对这种特定表面活性剂以及可能对其他化学刺激物的刺激反应的分子事件提供额外线索。
