文章基本信息

标题：Profiling Environmental Chemicals for Activity in the Antioxidant Response Element Signaling Pathway Using a High Throughput Screening Approach
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作者：Sunita J Shukla ; Ruili Huang ; Steven O Simmons 等
期刊名称：Environmental Health Perspectives
印刷版ISSN：0091-6765
电子版ISSN：1552-9924
出版年度：2012
卷号：120
期号：8
页码：1150-1156
DOI：10.1289/ehp.1104709
出版社：OCR Subscription Services Inc
摘要：Ba c k g r o u n d: Oxidative stress has been implicated in the pathogenesis of a variety of diseases ranging from cancer to neurodegeneration, highlighting the need to identify chemicals that can induce this effect. The antioxidant response element (ARE) signaling pathway plays an important role in the amelioration of oxidative stress. Thus, assays that detect the up-regulation of this path-way could be useful for identifying chemicals that induce oxidative stress.oB j e c t i v e s: We used cell-based reporter methods and informatics tools to efficiently screen a large collection of environmental chemicals and identify compounds that induce oxidative stress.Me t h o d s: We utilized two cell-based ARE assay reporters, b-lactamase and luciferase, to screen a U.S. National Toxicology Program 1,408-compound library (NTP 1408, which contains 1,340 unique compounds) for their ability to induce oxidative stress in HepG2 cells using quantitative high throughput screening (qHTS).re s u l t s: Roughly 3% (34 of 1,340) of the unique compounds demonstrated activity across both cell-based assays. Based on biological activity and structure–activity relationship profiles, we selected 50 compounds for retesting in the two ARE assays and in an additional follow-up assay that employed a mutated ARE linked to b-lactamase. Using this strategy, we identified 30 compounds that demonstrated activity in the ARE-bla and ARE-luc assays and were able to determine structural features conferring compound activity across assays.co n c l u s i o n s: Our results support the robustness of using two different cell-based approaches for identifying compounds that induce ARE signaling. Together, these methods are useful for prioritiz-ing chemicals for further in-depth mechanism-based toxicity testing
关键词：ARE; Nrf2; oxidative stress; qHTS; toxicity; Tox21