title: 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.
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.
authors.affiliation: Chair of Nutritional Toxicology, Institute of Nutritional Science, University of Potsdam, Germany.
authors.forename: Kristina
authors.initials: K
authors.lastname: Ullmann
authors.forename: Anne Maria
authors.initials: AM
authors.lastname: Wiencierz
authors.forename: Carsten
authors.initials: C
authors.lastname: Müller
authors.forename: Rene
authors.initials: R
authors.lastname: Thierbach
authors.forename: Andreas
authors.initials: A
authors.lastname: Steege
authors.forename: Shinya
authors.initials: S
authors.lastname: Toyokuni
authors.forename: Pablo
authors.initials: P
authors.lastname: Steinberg
mesh_terms: D000818:Animals; D016588:Anticarcinogenic Agents / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D001205:Ascorbic Acid / Q000494:pharmacology; D002273:Carcinogens / Q000494:pharmacology; D002331:Carnitine / Q000494:pharmacology; D002374:Catalase / Q000235:genetics* / Q000378:metabolism; D002392:Catechin / Q000031:analogs & derivatives / Q000494:pharmacology; D002478:Cells, Cultured; D002839:Chromans / Q000494:pharmacology; D006224:Cricetinae; D003412:Cricetulus; D019833:Genistein / Q000494:pharmacology; D005979:Glutathione Peroxidase / Q000235:genetics* / Q000378:metabolism; D006540:Herbicides / Q000494:pharmacology; D006801:Humans; D008156:Luciferases / Q000378:metabolism*; D010269:Paraquat / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000187:drug effects* / Q000235:genetics*; D011794:Quercetin / Q000494:pharmacology; D051381:Rats; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014803:Vitamin B Complex / Q000494:pharmacology
chemical_list: D016588:Anticarcinogenic Agents; D000975:Antioxidants; D002273:Carcinogens; D002839:Chromans; D006540:Herbicides; D014803:Vitamin B Complex; D002392:Catechin; D011794:Quercetin; C045651:epigallocatechin gallate; D019833:Genistein; D002374:Catalase; D005979:Glutathione Peroxidase; D008156:Luciferases; D013482:Superoxide Dismutase; D013755:Tetradecanoylphorbol Acetate; D010269:Paraquat; D001205:Ascorbic Acid; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid; D002331:Carnitine