Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 10 de 10
Filter
Add more filters










Publication year range
1.
Toxicol Sci ; 139(2): 293-300, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24675090

ABSTRACT

Mice are resistant to aflatoxin hepatotoxicity, primarily due to high expression of glutathione S-transferases (GSTs), and in particular the GSTA3 subunit. Nuclear factor erythroid 2 related factor 2 (Nrf2) signaling, which controls a broad-based cytoprotective response, was activated either genetically or pharmacologically in an attempt to rescue GSTA3 knockout mice from aflatoxin genotoxicity. Genetic activation of Nrf2 signaling was attained in a GSTA3: hepatocyte-specific Keap1 double knockout (DKO) mouse whereas pharmacologic activation of Nrf2 was achieved through pretreatment of mice with the triterpenoid 1-[2-cyano-3-,12-dioxoleana-1,9(11)-dien-28-oyl] imidazole (CDDO-Im) prior to aflatoxin B1 exposure. Following oral treatment with aflatoxin, urine was collected from mice for 24 h and hepatic and urinary aflatoxin metabolites then quantified using isotope dilution-mass spectrometry. Although Nrf2 was successfully activated genetically and pharmacologically, neither means affected the response of GSTA3 knockout mice to chemical insult with aflatoxin. Hepatic aflatoxin B1-N(7)-guanine levels were elevated 120-fold in GSTA3 knockout mice compared with wild-type and levels were not attenuated by the interventions. This lack of effect was mirrored in the urinary excretion of aflatoxin B1-N(7)-guanine. By contrast, urinary excretion of aflatoxin B1-N-acetylcysteine was >200-fold higher in wild-type mice compared with the single GSTA3 knockout or DKO mouse. The inability to rescue GSTA3 knockout mice from aflatoxin genotoxicity through the Nrf2 transcriptional program indicates that Gsta3 is unilaterally responsible for the detoxication of aflatoxin in mice.


Subject(s)
Aflatoxin B1/toxicity , Glutathione Transferase/genetics , Hypertension/drug therapy , Imidazoles/pharmacology , Mutagens/toxicity , NF-E2-Related Factor 2 , Oleanolic Acid/analogs & derivatives , Signal Transduction/drug effects , Adaptor Proteins, Signal Transducing/genetics , Aflatoxin B1/pharmacokinetics , Aflatoxin B1/urine , Animals , Cytoskeletal Proteins/genetics , Hepatocytes/drug effects , Hepatocytes/enzymology , Hepatocytes/metabolism , Hypertension/genetics , Hypertension/metabolism , Kelch-Like ECH-Associated Protein 1 , Liver/drug effects , Liver/enzymology , Liver/metabolism , Mice, Inbred C57BL , Mice, Knockout , Mutagens/pharmacokinetics , NF-E2-Related Factor 2/genetics , NF-E2-Related Factor 2/metabolism , Oleanolic Acid/pharmacology , Signal Transduction/genetics
2.
Eur J Pharmacol ; 620(1-3): 138-44, 2009 Oct 12.
Article in English | MEDLINE | ID: mdl-19698707

ABSTRACT

The synthetic oleanolic triterpenoid 1-[2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oyl]imidazole (CDDO-Imidazolide or CDDO-Im) is an extremely potent activator of Nrf2 signaling. In cells undergoing adipogenesis, CDDO-Im prevents lipid accumulation in an Nrf2-dependent manner. However, in vivo evidence for effects of CDDO-Im on obesity is lacking. The goals of these studies were to determine if CDDO-Im can prevent high-fat diet-induced obesogenesis in the mouse, and to elucidate the molecular target of drug action. Wild-type and Nrf2-disrupted C57BL/6J female mice were dosed 3 times per week with 30 micromol/kg CDDO-Im or vehicle by oral gavage, during 95 days of access to a control diet or a high-fat diet. Body weights, organ weights, hepatic fat accumulation and gene expression were measured. Treatment with CDDO-Im effectively prevented high-fat diet-induced increases in body weight, adipose mass, and hepatic lipid accumulation in wild-type mice but not in Nrf2-disrupted mice. Wild-type mice on a high-fat diet and treated with CDDO-Im exhibited higher oxygen consumption and energy expenditure than vehicle-treated mice, while food intake was lower in CDDO-Im-treated than vehicle-treated mice. Levels of gene transcripts for fatty acid synthesis enzymes were downregulated after CDDO-Im treatment in the liver of wild-type mice. This inhibitory effect of CDDO-Im on lipogenic gene expression was significantly reduced in Nrf2-disrupted mice. The results indicate that CDDO-Im is an exceedingly potent agent for preventing obesity, and identify the Nrf2 pathway as a novel target for management of obesogenesis.


Subject(s)
Anti-Obesity Agents/pharmacology , Dietary Fats/pharmacology , Imidazoles/pharmacology , NF-E2-Related Factor 2/metabolism , Obesity/chemically induced , Obesity/prevention & control , Oleanolic Acid/analogs & derivatives , Adipocytes/drug effects , Adipocytes/metabolism , Animals , Body Weight/drug effects , Energy Metabolism/drug effects , Fatty Acids/biosynthesis , Female , Gene Expression Regulation, Enzymologic/drug effects , Liver/drug effects , Liver/enzymology , Liver/metabolism , Mice , NF-E2-Related Factor 2/deficiency , Obesity/metabolism , Obesity/physiopathology , Oleanolic Acid/pharmacology , Oxidation-Reduction
3.
Carcinogenesis ; 30(6): 1024-31, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19386581

ABSTRACT

Loss of NF-E2-related factor 2 (Nrf2) signaling increases susceptibility to acute toxicity, inflammation and carcinogenesis in mice due to the inability to mount adaptive responses. In contrast, disruption of Keap1 (a cytoplasmic modifier of Nrf2 turnover) protects against these stresses in mice, although inactivating mutations in Keap1 have been identified recently in some human cancers. Global characterization of Nrf2 activation is important to exploit this pathway for chemoprevention in healthy, yet at-risk individuals and also to elucidate the consequences of hijacking the pathway in Keap1-mutant human cancers. Liver-targeted conditional Keap1-null, Albumin-Cre:Keap1((flox/-)) (CKO) mice provide a model of genetic activation of Nrf2 signaling. By coupling global gene expression analysis of CKO mice with analysis of pharmacologic activation using the synthetic oleanane triterpenoid 1-[2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oyl]imidazole (CDDO-Im), we are able to gain insight into pathways affected by Nrf2 activation. CDDO-Im is an extremely potent activator of Nrf2 signaling. CKO mice were used to identify genes modulated by genetic activation of Nrf2 signaling. The CKO response was compared with hepatic global gene expression changes in wild-type mice treated with CDDO-Im at a maximal Nrf2 activating dose. The results show that genetic and pharmacologic activation of Nrf2 signaling modulates pathways beyond detoxication and cytoprotection, with the largest cluster of genes associated with lipid metabolism. Genetic activation of Nrf2 results in much larger numbers of detoxication and lipid metabolism gene changes. Additionally, analysis of pharmacologic activation suggests that Nrf2 is the primary mediator of CDDO-Im activity, though other cell-signaling targets are also modulated following an oral dose of 30 micromol/kg.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Cytoskeletal Proteins/metabolism , Imidazoles/pharmacology , Liver/metabolism , NF-E2-Related Factor 2/metabolism , Oleanolic Acid/analogs & derivatives , Adaptor Proteins, Signal Transducing/genetics , Animals , Cytoskeletal Proteins/genetics , Kelch-Like ECH-Associated Protein 1 , Lipid Metabolism/drug effects , Male , Mice , Mice, Knockout , NF-E2-Related Factor 2/genetics , Oleanolic Acid/pharmacology , Oligonucleotide Array Sequence Analysis , Signal Transduction
4.
J Med Chem ; 50(23): 5824-32, 2007 Nov 15.
Article in English | MEDLINE | ID: mdl-17924616

ABSTRACT

Eight new side-chain allylic, benzylic, and propargylic ether analogs of the natural hormone calcitriol have been rationally designed and easily synthesized. Three of these 23-oxa ether analogs lacking the typical side-chain OH group are more antiproliferative in vitro and desirably less calcemic in vivo than the natural hormone. One of these three 23-oxa analogs has transcriptional potency almost as high as that of calcitriol, even though it binds to the human vitamin D receptor only about 1% as well as calcitriol.


Subject(s)
Calcitriol/analogs & derivatives , Calcitriol/chemical synthesis , Calcium/urine , Cell Proliferation/drug effects , Alkynes/chemical synthesis , Alkynes/pharmacology , Allyl Compounds/chemical synthesis , Allyl Compounds/pharmacology , Animals , Benzyl Compounds/chemical synthesis , Benzyl Compounds/pharmacology , Binding, Competitive , Calcitriol/pharmacology , Calcium Channels/biosynthesis , Calcium Channels/genetics , Cell Line , Duodenum/metabolism , Female , Humans , Keratinocytes/cytology , Keratinocytes/drug effects , Mice , Mice, Inbred C57BL , Rats , Receptors, Calcitriol/metabolism , Stereoisomerism , Steroid Hydroxylases/biosynthesis , Steroid Hydroxylases/genetics , Structure-Activity Relationship , TRPV Cation Channels/biosynthesis , TRPV Cation Channels/genetics , Transcription, Genetic/drug effects , Vitamin D3 24-Hydroxylase
5.
Int J Cancer ; 121(9): 1883-1891, 2007 Nov 01.
Article in English | MEDLINE | ID: mdl-17631644

ABSTRACT

Chronic inflammation has been associated with increased risk of developing cancer. The transcription factor NF-E2-related factor 2 (Nrf2) controls the expression of numerous antioxidative enzymes that have been shown to attenuate acute inflammation. The present study investigated the role of Nrf2 genotype in modulating inflammation-promoted colorectal tumorigenesis. Nrf2 wild-type (WT) and Nrf2-deficient (N0) mice were administered a single dose of azoxymethane followed by a 1-week dose of drinking water with or without 1% dextran sulfate sodium (DSS). Aberrant crypt foci were counted 3 weeks after the cessation of DSS treatment. DSS treatment significantly increased numbers of aberrant crypt foci in N0 mice, but not WT mice. The extent of inflammation over the course of DSS treatment was analyzed in both genotypes. Histological analysis of colon sections revealed that N0 mice had markedly increased inflammation and mucosal damage when compared to WT mice beginning on Day 6 of DSS treatment. Although similar levels of inflammatory and oxidative damage biomarkers were evident in colons from WT and N0 mice at the start of DSS treatment, increased colonic proinflammatory cytokine mRNA transcript levels, myeloperoxidase activity and 3-nitrotyrosine immunoreactivity were observed on Day 6 of DSS treatment in N0 mice, but not WT mice. Additionally, DSS treatment resulted in increased lipid peroxidation and loss of aconitase activity in N0 mice, but not WT mice, reflecting increased oxidative damage in colons from N0 mice. Taken together, these results clearly illustrate the role of Nrf2 in regulating an adaptive response that protects against early-phase inflammation-mediated tumorigenesis.


Subject(s)
Colitis/metabolism , Colitis/pathology , Colon/injuries , Colon/metabolism , Dextran Sulfate/pharmacology , NF-E2-Related Factor 2/deficiency , NF-E2-Related Factor 2/metabolism , Animals , Azoxymethane/pharmacology , Biomarkers , Colitis/chemically induced , Colitis/genetics , Colon/drug effects , Colon/pathology , Cytokines/genetics , Female , Gene Expression Regulation , Mice , Mice, Knockout , NF-E2-Related Factor 2/genetics , Oxidative Stress/drug effects
6.
Bioorg Med Chem ; 15(16): 5509-18, 2007 Aug 15.
Article in English | MEDLINE | ID: mdl-17560112

ABSTRACT

Novel fluorinated sulfone analogs of the hormone 1alpha,25-dihydroxyvitamin D(3) have been designed and synthesized in order to study the biological effects of fluorine incorporation at the terminus of the C,D-ring side chain. Although biologically active 26,27-hexafluorinated 1alpha,25-dihydroxyvitamin D(3) analogs have been synthesized previously, this investigation reports the first successful fluorinated series in which trifluoromethyl sulfone analogs present a favorable biological profile. This study shows that two new analogs featuring incorporation of a synthetically simple single trifluoromethyl sulfone group have significantly increased antiproliferative activity while causing desirably low in vivo calciuria relative to that of calcitriol. Incorporation of additional fluorines, as in a perfluorobutyl analog, results in a loss of antiproliferative activity.


Subject(s)
Calcitriol/chemistry , Fluorine/chemistry , Sulfones/chemistry , Vitamin D/analogs & derivatives , Animals , Calcium/urine , Cell Proliferation/drug effects , Molecular Structure , Rats , Vitamin D/chemical synthesis , Vitamin D/chemistry , Vitamin D/pharmacology
7.
J Med Chem ; 47(27): 6854-63, 2004 Dec 30.
Article in English | MEDLINE | ID: mdl-15615534

ABSTRACT

A dozen 24-sulfoximine analogues of the hormone 1alpha,25-dihydroxyvitamin D(3) were prepared, differing not only at the stereogenic sulfoximine stereocenter but also at the A-ring. Although these sulfoximines were not active transcriptionally and were only very weakly antiproliferative, some of them are powerful hydroxylase enzyme inhibitors. Specifically, 24-(S)-NH phenyl sulfoximine 3a is an extremely potent CYP24 inhibitor (IC(50) = 7.4 nM) having low calcemic activity. In addition, this compound shows high selectivity toward the CYP24 enzyme in comparison to CYP27A1 (IC(50) > 1000 nM) and CYP27B (IC(50) = 554 nM).


Subject(s)
Calcitriol/analogs & derivatives , Calcium/urine , Cytochrome P-450 Enzyme Inhibitors , Steroid Hydroxylases/antagonists & inhibitors , Sulfones/chemical synthesis , Animals , Calcitriol/pharmacology , Male , Rats , Rats, Inbred F344 , Structure-Activity Relationship , Sulfones/pharmacology , Vitamin D3 24-Hydroxylase
9.
Carcinogenesis ; 24(3): 461-7, 2003 Mar.
Article in English | MEDLINE | ID: mdl-12663505

ABSTRACT

The cancer chemopreventive actions of oltipraz (4-methyl-5-[2-pyrazinyl]-1,2-dithiole-3-thione) have been primarily associated with the induction of phase 2 detoxifying enzymes through transcriptional activation of the antioxidant response element (ARE) in the promoter regions of these genes. The transcription factor Nrf2 has been shown to bind to and activate AREs. Previously, we demonstrated that nrf2-deficient mice had low basal expression of phase 2 enzymes and were substantially more susceptible to benzo[a]pyrene (B[a]P)-induced neoplasia of the forestomach than wild-type. Moreover, loss of Nrf2 abrogated the chemopreventive action of oltipraz, when administered 48 h before B[a]P, an interval allowing maximal induction of many phase 2 enzymes. Oltipraz also inhibits some cytochrome P450s involved in the bioactivation of B[a]P. In the present study we observed that oltipraz had no protective effect on tumor burden in the forestomach of nrf2-deficient mice when administered 1 h before B[a]P, a timeline that selectively optimizes for possible inhibitory effects on cytochrome P450s. To evaluate the role of nrf2 genotype on B[a]P disposition, levels of B[a]P-DNA adducts were measured as tetrols released from DNA isolated from target (forestomach) and non-target tissues (liver) of wild-type and nrf2-deficient mice treated with either vehicle or oltipraz 1 or 48 h before B[a]P. Levels of B[a]P-DNA adducts in forestomach were significantly higher in nrf2-deficient compared with wild-type mice. Oltipraz treatment at 1 or 48 h before B[a]P had no protective effect on forestomach tetrol levels in nrf2-deficient mice, whereas a significant reduction was observed in wild-type mice treated with oltipraz 48 h, but not 1 h, before carcinogen. Combining all treatments and genotypes, there was a strong correlation (R(2) = 0.91) between levels of B[a]P-DNA adducts in forestomach and subsequent yield of tumors. In contrast to the results in forestomach, nrf2 genotype did not modify hepatic B[a]P-DNA adduct levels while both oltipraz treatments were protective, suggesting that Nrf2-independent mechanisms (e.g. P450 inhibition) for oltipraz can also occur in vivo in some tissues.


Subject(s)
Anticarcinogenic Agents/pharmacology , DNA Adducts/drug effects , DNA Adducts/genetics , DNA-Binding Proteins/genetics , Genotype , Pyrazines/pharmacology , Trans-Activators/genetics , Animals , Benzo(a)pyrene , Mice , Mice, Knockout , NF-E2-Related Factor 2 , Thiones , Thiophenes
10.
Proc Natl Acad Sci U S A ; 99(11): 7610-5, 2002 May 28.
Article in English | MEDLINE | ID: mdl-12032331

ABSTRACT

Gastric infection with Helicobacter pylori is a cosmopolitan problem, and is especially common in developing regions where there is also a high prevalence of gastric cancer. These infections are known to cause gastritis and peptic ulcers, and dramatically enhance the risk of gastric cancer. Eradication of this organism is an important medical goal that is complicated by the development of resistance to conventional antimicrobial agents and by the persistence of a low level reservoir of H. pylori within gastric epithelial cells. Moreover, economic and practical problems preclude widespread and intensive use of antibiotics in most developing regions. We have found that sulforaphane [(-)-1-isothiocyanato-(4R)-(methylsulfinyl)butane], an isothiocyanate abundant as its glucosinolate precursor in certain varieties of broccoli and broccoli sprouts, is a potent bacteriostatic agent against 3 reference strains and 45 clinical isolates of H. pylori [minimal inhibitory concentration (MIC) for 90% of the strains is

Subject(s)
Anticarcinogenic Agents/pharmacology , Helicobacter pylori/drug effects , Stomach Neoplasms/prevention & control , Thiocyanates/pharmacology , Agaricales , Amoxicillin/pharmacology , Animals , Benzo(a)pyrene , Brassica , Clarithromycin/pharmacology , Drug Resistance, Microbial , Female , Helicobacter pylori/growth & development , Helicobacter pylori/isolation & purification , Humans , Isothiocyanates , Metronidazole/pharmacology , Mice , Mice, Inbred ICR , Phytotherapy , Stomach Neoplasms/chemically induced , Stomach Neoplasms/microbiology , Sulfoxides
SELECTION OF CITATIONS
SEARCH DETAIL
...