Activity of the dietary antioxidant ergothioneine in a virus gene-based assay for inhibitors of HIV transcription.
Activity of the dietary antioxidant ergothioneine in a virus gene-based assay for inhibitors of HIV transcription.
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膳食抗氧化剂麦角硫因在基于病毒基因的 HIV 转录抑制剂测定中的活性。
DOI：
10.1002/biof.5520270114
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Taylor,EthanWill
中科院分区：
文献类型：
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作者：
Xiao,Lianchun;Zhao,Lijun;Li,Ting;Hartle,DianeK;Aruoma,OkezieI;Taylor,EthanWill
HIV-1的“长末端重复序列”（LTR）是细胞转录因子（如NF-κB）的靶标，并在整合到宿主DNA中时充当病毒基因组的启动子-增强子。各种LTR-报告基因构建体已用于HIV-1转录的激活剂或抑制剂的体外研究，例如，显示抗氧化剂如硫辛酸和硒抑制NF-κB-依赖性HIV-1 LTR活化。一种这样的构建体是pHIVlacZ质粒，其中HIV-1 LTR驱动lacZ基因（编码β-半乳糖苷酶，β-gal）的表达。通常，对于抑制剂筛选，使用肿瘤坏死因子-α（TNF-α）激活用pHIVlacZ转染的细胞，并使用比色邻硝基苯酚测定来评估β-gal活性的变化。如本文所述，开发了该检测方法的一种变体，其中LTR激活由pro-fs诱导，pro-fs是一种通过蛋白酶基因的−1移码编码的新型HIV-1基因产物。与对照组相比，用pHIVlacZ沿着与pro-fs构建体共转染细胞产生了β-gal活性的显著增加。L-麦角硫因剂量依赖性抑制TNF-α-介导和pro-fs-介导的β-gal活性增加，IC 50约为6 mM。因此，涉及来自食用植物的麦角硫因的抗氧化策略可能对慢性免疫缺陷疾病有益。
The “Long Terminal Repeat” (LTR) of HIV‐1 is the target of cellular transcription factors such as NF‐κB, and serves as the promoter‐enhancer for the viral genome when integrated in host DNA. Various LTR‐reporter gene constructs have been used forin vitrostudies of activators or inhibitors of HIV‐1 transcription, e.g., to show that antioxidants such as lipoic acid and selenium inhibit NF‐κB‐dependent HIV‐1 LTR activation. One such construct is the pHIVlacZ plasmid, with the HIV‐1 LTR driving expression of the lacZ gene (encoding β‐galactosidase, β‐gal). Typically, for inhibitor screening, cells transfected with pHIVlacZ are activated using tumor necrosis factor‐α (TNF‐α), and the colorimetric o‐nitrophenol assay is used to assess changes in β‐gal activity. A variant of this assay was developed as described here, in which LTR activation was induced by pro‐fs, a novel HIV‐1 gene product encoded via a −1 frameshift from the protease gene. Cotransfection of cells with pHIVlacZ along with a pro‐fs construct produced a signifcant increase in β‐gal activity over controls. L‐ergothioneine dose dependently inhibited both TNF‐α‐mediated and pro‐fs‐mediated increases in β‐gal activity, with an IC50of about 6 mM. Thus antioxidant strategy involving ergothioneine derived from food plants might be of benefit in chronic immunodeficiency diseases.