A high-throughput reporter gene assay to prove the ability of natural compounds to modulate glutathione peroxidase,superoxide dismutase and catalase gene promoters in V79 cells |
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| Authors: | Kristina Ullmann Anne Maria Wiencierz Carsten Müller René Thierbach Andreas Steege Shinya Toyokuni |
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| Institution: | 1. Chair of Nutritional Toxicology, Institute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114-116, 14558 Nuthetal, Germany;2. Institute for Food Toxicology and Analytical Chemistry, University of Veterinary Medicine Hannover, Bischofsholer Damm 15, 30173 Hannover, Germany;3. Institut für Vegetative Physiologie, Charité-Universit?tsmedizin Berlin, 10117 Berlin, Germany;4. Department of Pathology and Biology of Diseases, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan;5. Department of Pathology and Biological Responses, Graduate School of Medicine, Nagoya University, Nagoya 466-8550, Japan |
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| Abstract: | The aim of the study was to establish a 96-well microtiter plate-based reporter gene assay to test the influence of natural compounds on the promoter activities of rat catalase, human glutathione peroxidase and human superoxide dismutase expressed in V79 cells. Luciferase expression vectors with the promoter regions of the genes coding for the three above-mentioned enzymes were constructed and transfected into V79 cells. Thereafter the ability of sodium ascorbate, L-carnitine, catechin, epigallocatechin gallate, genistein, paraquat, quercetin, 12-O-tetradecanoylphorbol-13-acetate and Trolox to enhance the promoter activities was evaluated. Genistein, paraquat and quercetin led to a statistically significant increase in the glutathione peroxidase and superoxide dismutase gene promoter activities. None of the compounds tested enhanced the catalase gene promoter activity. The reporter gene assay described in this report is easy to perform, fast and allows one to test a high number of compounds and different concentrations of a single compound at the same time. |
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| Keywords: | Catalase glutathione peroxidise superoxide dismutase reporter gene assay natural compounds |
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