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.
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.
Ullmann
Kristina
K
Chair of Nutritional Toxicology, Institute of Nutritional Science, University of Potsdam, Germany.
Wiencierz
Anne Maria
AM
Müller
Carsten
C
Thierbach
Rene
R
Steege
Andreas
A
Toyokuni
Shinya
S
Steinberg
Pablo
P
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
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