α‐Lipoic acid blocks HIV‐1 LTR‐dependent expression of hygromycin resistance in THP‐1 stable transformants

FEBS Letters

Matsuyama, Mototaka³  Merin, Jocelyn P.³  Baba, Masanori²  Kira, Toshihiko¹  Okamoto, Takashi³ 

[1] Department of Microbiology, Fukushima Medical College, Fukushima 960-12, Japan;Division of Human Retroviruses, Center for Chronic Viral Diseases, Faculty of Medicine, Kagoshima University, Kagoshima 890, Japan;Department of Molecular Genetics, Nagoya City University Medical School, Nagoya 467, Japan

关键词: Gene expression;    Reactive oxygen intermediate;    Nuclear factor-κB;    α-Lipoic acid;    N-Acetylcysteine;    HIV-1;    AIDS;    ROI;    reactive oxygen intermediates;    NF-κB;    nuclear factor-κB;    LA;    α-lipoic acid;    DHLA;    dihydrolipoate;    NAC;    N-acetylcysteine;    HIV-1;    human immunodeficiency virus type 1;    LTR;    long terminal repeat;    GSH;    reduced glutathione;    MTT;    3(4;    5-dimethylthiazol-2-yl)-2;    5-diphenyl-tetrazolium bromide;    TNF-α;    tumor necrosis factor-α;   

DOI  :  10.1016/0014-5793(96)00919-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Gene expression of human immunodeficiency virus (HIV) depends on a host cellular transcription factors including nuclear factor-κB (NF-κB). The involvement of reactive oxygen intermediates (ROI) has been implicated as intracellular messengers in the inducible activation of NF-κB. In this study, we compared the efficacy of two antioxidants, α-lipoic acid (LA) and N-acetylcysteine (NAC), which are widely recognized NF-κB inhibitors. Here, we demonstrate that LA has a more potent activity in inhibiting NF-κB-mediated gene expression in THP-1 cells that have been stably transfected with a plasmid bearing a hygromycin B resistance gene under the control of HIV-1 long terminal repeat (LTR) promoter. The spontaneous activation of NF-κB in this cell culture system leads to expression of the hygromycin phosphotransferase gene hence rendering the cells resistance to hygromycin B. In this study, the effect of the test compounds against transcriptional activity of HIV-1 LTR was evaluated based on the degree of cellular toxicity due to the inhibitory activity on the expression of hygromycin B resistance gene in the presence of hygromycin B. We also found that 0.2 mM LA could cause 40% reduction in the HIV-1 expression from the TNF-α-stimulated OM 10.1, a cell line latently infected with HIV-1. On the other hand, 10 mM NAC was required to elicit the same effect. Furthermore, the initiation of HIV-1 induction by TNF-α was completely abolished by 1 mM LA. These findings confirm the involvement of ROI in NF-κB-mediated HIV gene expression as well as the efficacy of LA as a therapeutic regimen for HIV infection and acquired immunodeficiency syndrome (AIDS). Moreover, this study validates the applicability of our present assay system which we primarily designed for the screening of candidate drugs against HIV-1 gene expression.

【 授权许可】

Unknown