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使用DNA微阵列在重建的人表皮模型上评估皮肤刺激性。

Evaluation of the skin irritation using a DNA microarray on a reconstructed human epidermal model.

作者信息

Niwa Makoto, Nagai Kanji, Oike Hideaki, Kobori Masuko

机构信息

Katakura Chikkarin Co., Ltd, Ibaraki, Japan.

出版信息

Biol Pharm Bull. 2009 Feb;32(2):203-8. doi: 10.1248/bpb.32.203.

Abstract

To avoid the need to use animals to test the skin irritancy potential of chemicals and cosmetics, it is important to establish an in vitro method based on the reconstructed human epidermal model. To evaluate skin irritancy efficiently and sensitively, we determined the gene expression induced by a topically-applied mild irritant sodium dodecyl sulfate (SDS) in a reconstructed human epidermal model LabCyte EPI-MODEL (LabCyte) using a DNA microarray carrying genes that were related to inflammation, immunity, stress and housekeeping. The expression and secretion of IL-1alpha in reconstructed human epidermal culture is known to be induced by irritation. We detected the induction of IL-1alpha expression and its secretion into the cell culture medium by treatment with 0.075% SDS for 18 h in LabCyte culture using DNA microarray, quantitative reverse-transcription polymerase chain reaction (RT-PCR) and ELISA. DNA microarray analysis indicated that the expression of 10 of the 205 genes carried on the DNA microarray was significantly induced in a LabCyte culture by 0.05% or 0.075% SDS irritation for 18 h. RT-PCR analysis confirmed that SDS treatment significantly induced the expressions of interleukin-1 receptor antagonist (IL-1RN), FOS-like antigen 1 (FOSL1), heat shock 70 kDa protein 1A (HSPA1) and myeloid differentiation primary response gene (88) (MYD88), as well as the known marker genes for irritation IL-1beta and IL-8 in a LabCyte culture. Our results showed that a DNA microarray is a useful tool for efficiently evaluating mild skin irritation using a reconstructed human epidermal model.

摘要

为避免使用动物来测试化学品和化妆品的皮肤刺激性,建立基于重建人表皮模型的体外方法很重要。为了高效且灵敏地评估皮肤刺激性,我们使用携带与炎症、免疫、应激和管家基因相关的基因的DNA微阵列,在重建人表皮模型LabCyte EPI - MODEL(LabCyte)中测定了局部应用轻度刺激性十二烷基硫酸钠(SDS)诱导的基因表达。已知在重建人表皮培养物中,IL - 1α的表达和分泌可由刺激诱导。我们在LabCyte培养物中,通过DNA微阵列、定量逆转录聚合酶链反应(RT - PCR)和酶联免疫吸附测定(ELISA),检测了用0.075% SDS处理18小时后IL - 1α表达的诱导及其向细胞培养基中的分泌。DNA微阵列分析表明,在LabCyte培养物中,DNA微阵列上携带的205个基因中的10个基因的表达在0.05%或0.075% SDS刺激18小时后被显著诱导。RT - PCR分析证实,SDS处理在LabCyte培养物中显著诱导了白细胞介素 - 1受体拮抗剂(IL - 1RN)、FOS样抗原1(FOSL1)、热休克70 kDa蛋白1A(HSPA1)和髓样分化初级反应基因(88)(MYD88)的表达,以及已知的刺激性标记基因IL - 1β和IL - 8的表达。我们的结果表明,DNA微阵列是使用重建人表皮模型有效评估轻度皮肤刺激的有用工具。

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