Transcriptomics was performed to gain insight into mechanisms of food additives butylated hydroxytoluene (BHT), curcumin (CC), propyl gallate (PG), and thiabendazole (TB), additives for which interactions in the liver can not be excluded. Additives were administered in diets for 28 days to Sprague-Dawley rats and cDNA microarray experiments were performed on hepatic RNA. BHT induced changes in the expression of 10 genes, including phase I (CYP2B1/2; CYP3A9; CYP2C6) and phase II metabolism (GST μ2). The CYP2B1/2 and GST expression findings were confirmed by real time RT-PCR, western blotting, and increased GST activity towards DCNB. CC altered the expression of 12 genes. Three out of these were related to peroxisomes (phytanoyl-CoA dioxygenase, enoyl-CoA hydratase; CYP4A3). Increased cyanide insensitive palmitoyl-CoA oxidation was observed, suggesting that CC is a weak peroxisome proliferator. TB changed the expression of 12 genes, including CYP1A2. In line, CYP1A2 protein expression was increased. The expression level of five genes, associated with p53 was found to change upon TB treatment, including p53 itself, GADD45α, DN-7, protein kinase C β and serum albumin. These array experiments led to the novel finding that TB is capable of inducing p53 at the protein level, at least at the highest dose levels employed above the current NOAEL. The expression of eight genes changed upon PG administration. This study shows the value of gene expression profiling in food toxicology in terms of generating novel hypotheses on the mechanisms of action of food additives in relation to pathology. © 2008 Elsevier Ltd. All rights reserved. Molecular Sequence Numbers: GENBANK: AA818024, AA818192, AA818350, AA818412, AA818680, AA818833, AA818896, AA818960, AA819087, AA858732, AA858938, AA858966, AA859305, AA875380, AA876052, AA899808, AA899815, AA900472, AA900486, AA924591, AA926032, AA957616, AA963084, AA964507, AA964615, AA964751, AA997188, AA997994, AA999001, AI029442, AI029652, AI044024, AI044091, AI045500, AI059505, AI059951, AI060206, AI070295, AI070587, AI136404; Chemicals / CAS: butylcresol, 128-37-0, 30587-81-6; curcumin, 458-37-7; cyanide, 57-12-5; cytochrome P450 2B1, 330207-10-8; cytochrome P450 2B2, 331462-97-6; cytochrome P450, 9035-51-2; enoyl coenzyme A hydratase, 9027-13-8; gallic acid propyl ester, 121-79-9; glutathione transferase, 50812-37-8; palmitoyl coenzyme A, 1763-10-6; RNA, 63231-63-0; serum albumin, 9048-46-8; tiabendazole, 148-79-8; Aryl Hydrocarbon Hydroxylases, EC 1.14.14.1; Butylated Hydroxytoluene, 128-37-0; Curcumin, 458-37-7; Cytochrome P-450 CYP1A2, EC 1.14.14.1; Cytochrome P-450 CYP2B1, EC 1.14.14.1; DNA, Complementary; Food Additives; Glutathione Transferase, EC 2.5.1.18; Palmitoyl Coenzyme A, 1763-10-6; Propyl Gallate, 121-79-9; steroid 16-beta-hydroxylase, EC 1.14.14.1; Steroid Hydroxylases, EC 1.14.-; Thiabendazole, 148-79-8