This research demonstrated the protective effect and possible mechanism of the Sophora viciifolia extract (SVE) against acetaminophen-induced liver injury in mice. The levels of ALT and AST in the serum and antioxidant enzyme activity in the liver were measured. We used immunohistochemistry to detect CYP2E1, Nrf2, and Keap1 protein expression in the liver. The mRNA expression in the liver of TNF-α, NF-κB, and IL-6, Nrf2 and its downstream genes HO-1 and GCLC were measured by qRT-PCR. We found that SVE could decrease the ALT and AST levels, promote the activities of SOD, CAT, GSH-Px, and GSH, and ameliorate pathological liver lesions. SVE could down-regulate the mRNA expression of inflammatory factors and up-regulate Nrf2, HO-1 and GCLC. SVE reduced the protein expression of the CYP2E1 and increased the Nrf2 and Keap1. SVE has been shown to have a protective effect against APAP-induced liver injury, possibly through activation of the Keap1-Nrf2 pathway.

• MeSH
• Antioxidants pharmacology   MeSH
• Chemical and Drug Induced Liver Injury, Chronic * MeSH
• Cytochrome P-450 CYP2E1 genetics   metabolism   MeSH
• NF-E2-Related Factor 2 genetics   metabolism   MeSH
• Kelch-Like ECH-Associated Protein 1 genetics   metabolism   MeSH
• RNA, Messenger MeSH
• Mice MeSH
• Fruit metabolism   MeSH
• Acetaminophen * adverse effects   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

• MeSH
• Circadian Clocks MeSH
• Circadian Rhythm * MeSH
• Cytochrome P-450 CYP2E1 MeSH
• Gene Expression MeSH
• Transcription, Genetic MeSH
• Hepatocytes MeSH
• Drug Chronotherapy MeSH
• Drug Interactions MeSH
• Humans MeSH
• Acetaminophen adverse effects   MeSH
• Patient-Centered Care MeSH
• CLOCK Proteins MeSH
• ARNTL Transcription Factors MeSH
• Check Tag
• Humans MeSH

The effects of clotrimazole (CLO) and dexamethasone (DEX), both detected in the aquatic environment, were assessed on inhibition of cytochrome P450 (CYP450) in hepatic microsomes of rainbow trout. Activity of three CYP450 isoforms: ethoxyresorufin O-deethylase (EROD; CYP1A), 7-benzyloxy-4-trifluoromethylcoumarin O-debenzylase (BFCOD; CYP3A) and p-nitrophenol hydroxylase (PNPH; CYP2E1-like protein) was investigated in the presence of four concentrations of CLO and DEX. Clotrimazole in a concentration range of 1-100μM decreased the activity of EROD and BFCOD. The inhibition was reversible, as pre-incubation of the microsomes with CLO, before addition of the substrate, had no effect. EROD activity was non-competitively inhibited with a Ki of 0.5μM, and BFCOD activity revealed competitive inhibition with a Ki of 0.04μM. The relatively low Ki for CLO inhibition of EROD and BFCOD activity may indicate that the ability of CYP1A and CYP3A to metabolize xenobiotics is reduced in the presence of CLO. PNPH activity was not affected by CLO. DEX showed no inhibitory potency on any investigated reaction. CLO, but not DEX, inhibited EROD and BFCOD activity by different mechanisms.

• MeSH
• Cytochrome P-450 CYP1A1 antagonists & inhibitors   metabolism   MeSH
• Cytochrome P-450 CYP2E1 metabolism   MeSH
• Cytochrome P-450 CYP3A metabolism   MeSH
• Dexamethasone chemistry   pharmacology   MeSH
• Cytochrome P-450 CYP2E1 Inhibitors MeSH
• Cytochrome P-450 CYP3A Inhibitors MeSH
• Cytochrome P-450 Enzyme Inhibitors MeSH
• Microsomes, Liver drug effects   enzymology   MeSH
• Kinetics MeSH
• Clotrimazole chemistry   pharmacology   MeSH
• Oncorhynchus mykiss metabolism   MeSH
• Cytochrome P-450 Enzyme System metabolism   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Results of pilot study suggested co-segregation of genotypes of biotransformation enzymes with higher risk of lymphoma.Authors would like to focus to case-control study of patients with nonHodgkin´s lymphomas (NHL) and compare DNA from blood and tumors.Frequency of polymorphisms in: CYP1B1, CYP2E1, EPHX, NQO1,GSTM1/P1/Tl, and p532 will be assessed by PCR-RFLP in DNA from white blood cells. In DNA samples from NHL tumors, mutations in NBS1 and BCL-6 will be followed by fragment analysis and sequencing. Collaborating surgeons will provide information about patient staging, histopathological type,grading and size of tumor, nodal status,presence of metastases, biochemical markers, character and result of chemotherapy, overall and disease-free survival. Thefollowed biologic markers of structural and functional DNA damage may help to identify persons at risk, assist in prognostic predictions, and provide baselines to assess efficiacy of therapeutic interventions for treatment and management of NHL.

Výsledky pilotní studie naznačily vztahy mezi genotypy biotransformačních enzymů a vyšším rizikem vzniku lymfomů. Autoři plánují soustředit výzkum na srovnání pacientů s neHodgkinskými lymfomy(NHL) se zdravými kontrolami a vzorků krve s tumorovou tkání.DNA z bílých krvinek budou pomocí metody PCR-RFLP stanoveny frekvence výskytu genetických polymorfismů: CYP1B1, CYP2E1,EPHX,NQO1,GSTM1/p1/T1 a p53. Ve zvorcích tumorů budou fragmentační analýzou a sekvenováním studovány mutace v genech NBS1 a BCL6. Kolektiv lékařů poskytne informace o pacientu a průběhu onemocnění, např.:staging,histologický typ,grading a velikost tumoru, zasažení uzlin, výskyt metastáz, biochemické markery, typ a výsledek chemoterapie, celkové a bezpříznakové přežívání. Sledované biologické markery strukturního a funkčního poškození DNA by mohly přispět k identifikaci osob s rizikem NHL, prognostickým predikcím a k posouzení účinnosti potenciálních léčebných a výživových intervencí v prevenci, léčbě a zvládání nemocí.

• MeSH
• Biotransformation MeSH
• Enzymes MeSH
• Molecular Mechanisms of Pharmacological Action MeSH
• Lymphoma, Non-Hodgkin diagnosis   therapy   MeSH
• Risk Factors MeSH
• Genes, Tumor Suppressor MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• hematologie a transfuzní lékařství
• biologie
• onkologie
• NML Publication type
• závěrečné zprávy o řešení grantu IGA MZ ČR

Ethanol is linked to several pathologies like alcohol liver injury, neurotoxicity, cardiomyopathy, fetal alcoholic syndrome or cancer. It is generally accepted that oxidative stress plays a central role in their pathogenesis. After chronic and excessive consumption, alcohol may accelerate oxidative mechanisms both directly via increased production of reactive oxygen species and indirectly by impairing protective mechanisms against them. Ethanol, its metabolites arising during its metabolic degradation as well as novel compounds formed via ethanol induced oxidative stress, especially during the action of the ethanol inducible microsomal cytochrome CYP2E1, may apart from direct damage to biological structures affect signal transduction pathways thus modulating and potentiating damage. Alteration of the redox status of cells following chronic ethanol misuse may have profound effects on cellular function and viability and lead to cell death and tissue damage. These changes linked to pathologic processes in the organism, are related to alteration of intracellular signaling pathways associated with protein kinases and transcription factor activation. Mainly mitogen activated protein kinase (MAPK) family, transcription factors-nuclear factor kappaB (NF-kappaB) and activating protein 1 (AP-1) are involved in the deterioration of cells and organs. The response is cell-type specific and depends on the dose of ethanol. Oxido-reduction balance, regulatory disturbances and signal transduction cascades responsible for alcoholic damage have been partially described, nevertheless, further studies are required to allow future novel diagnostic and therapeutical strategies. We are only at the beginning ...

• MeSH
• Liver Diseases, Alcoholic metabolism   MeSH
• Ethanol metabolism   MeSH
• Financing, Organized MeSH
• Liver metabolism   MeSH
• Kupffer Cells enzymology   MeSH
• Humans MeSH
• Mitogen-Activated Protein Kinases metabolism   MeSH
• NF-kappa B metabolism   MeSH
• Oxidative Stress genetics   MeSH
• Signal Transduction physiology   MeSH
• Tumor Necrosis Factor-alpha metabolism   MeSH
• Activating Transcription Factor 1 metabolism   MeSH
• Check Tag
• Humans MeSH

Genetic polymorphisms influence the metabolism of ethanol and methanol, but the potential effects of genetic predisposition on the clinical course, outcome and short-term health sequelae of acute methanol poisoning are unknown. To evaluate the role of the MEOS system in methanol poisoning, we analysed the effect of three polymorphisms (RsaI - rs2031920; PstI - rs3813867; insertion/deletion I/D) within the CYP2E1 enzyme (MEOS system) in 50 adult survivors of methanol poisoning and compared their genotype frequencies with 460 controls. The minor allele frequencies of all three polymorphisms were below 5% in both groups. We did not detect significant differences in the genotype frequencies between survivors of methanol poisoning and controls (p = 0.34 for the RsaI variant; p = 0.59 for the PstI variant and p = 0.21 for the I/D polymorphism). The carriers of at least one minor allele in the CYP2E1 gene had less severe clinical symptoms and better short-term outcome after acute poisoning. Variants within the CYP2E1 gene are likely not significant genetic determinants of acute methanol poisoning (if survivors are analysed), but they may influence the severity of methanol poisoning and its visual/central nervous system (CNS) outcome.

• MeSH
• Alleles MeSH
• Cytochrome P-450 CYP2E1 genetics   MeSH
• Adult MeSH
• Gene Frequency MeSH
• Genetic Predisposition to Disease MeSH
• Genotype MeSH
• Middle Aged MeSH
• Humans MeSH
• Methanol poisoning   MeSH
• Young Adult MeSH
• Polymorphism, Genetic * MeSH
• Survivors MeSH
• Aged MeSH
• Severity of Illness Index MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Two non-symmetric bispyridine oxime - based reactivators of acetylcholinesterase enzyme (AChE), labeled as K027 (1-(4-carbamoylpyridinium)-3-(4-hydroxyiminomethylpyridinium)-propane dibromide) and K203 ((E)-1-(4- carbamoylpyridinium)-4-(4-hydroxyiminomethylpyridinium)-but-2-ene dibromide) were tested for their potential to inhibit activities of human liver microsomal cytochromes P450 (CYP). Both oximes are very potent reactivators of organophosphate-inhibited AChE. An interaction of both compounds with CYP in human liver microsomal preparation was detected using difference spectroscopy. The compounds were shown to bind to CYP enzymes with spectral binding constants of 5.04 ± 1.79 nM (K027) and 5.2 ± 2.6 nM (K203). Enzymology studies were subsequently performed aimed at determining which of the nine most important CYP involved in drug is affected by this interaction. The results have shown no prominent inhibition of individual CYP activities with either compounds except in the case of CYP2E1 and K203. Diagnostic Dixon plot revealed that K203 acted as an uncompetitive inhibitor of CYP2E1. Inhibition of this activity however is not as prominent as to make a potent drug interaction likely. Hence, the interaction of K027 and K203 oxime-type AChE reactivators with human liver microsomal CYP enzymes does not seem to be of prominent clinical importance and both compounds could be safely used in this respect as antidotes with low risk of drug interactions.

• MeSH
• Acetylcholinesterase metabolism   MeSH
• Cytochrome P-450 Enzyme Inhibitors pharmacology   MeSH
• Microsomes, Liver enzymology   MeSH
• Humans MeSH
• Molecular Structure MeSH
• Oximes pharmacology   MeSH
• Pyridinium Compounds pharmacology   MeSH
• Cholinesterase Reactivators pharmacology   MeSH
• Cytochrome P-450 Enzyme System metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Cytochromes P450 2E1 of human and minipig origin were examined by absorption spectroscopy under high hydrostatic pressure and by resonance Raman spectroscopy. Human enzyme tends to denature to the P420 form more easily than the minipig form; moreover, the apparent compressibility of the heme active site (as judged from a redshift of the absorption maximum with pressure) is greater than that of the minipig counterpart. Relative compactness of the minipig enzyme is also seen in the Raman spectra, where the presence of planar heme conformation was inferred from band positions characteristic of the low-spin heme with high degree of symmetry. In this respect, the CYP2E1 seems to be another example of P450 conformational heterogeneity as shown, e.g., by Davydov et al. for CYP3A4 [Biochem. Biophys. Res. Commun. 312 (2003) 121-130]. The results indicate that the flexibility of the CYP active site is likely one of its basic structural characteristics.

• MeSH
• Cytochrome P-450 CYP2E1 chemistry   metabolism   MeSH
• Financing, Organized MeSH
• Humans MeSH
• Swine MeSH
• Spectrum Analysis, Raman methods   MeSH
• Spectrophotometry methods   MeSH
• Binding Sites MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Comparative Study MeSH

The prevalence of obesity and other obesity-related diseases is increasing worldwide. Obesity is a disease characterized by increased body weight, or a condition resulting from excessive accumulation of body fat. Due to increased body fat deposits, obesity has also been associated with increased mortality resulting from higher incidence rates of hypertension, diabetes, or various types of cancer, such as breast, colorectal, cervical and prostate cancer. Physiological changes associated with obesity are likely to result in altered drug biotransformation. The main enzymes enabling the oxidative biotransformation of most drugs are cytochromes P450 (CYPs). The review summarizes how pathophysiological factors, especially obesity, affect properties (e.g. enzyme activity, protein expression, gene expression) of CYP enzymes in various experimental models of human obesity. Results reported by various authors suggest that obesity is associated with a decrease of CYP activities (except for the CYP2C and CYP2E1 enzymes). The only exception is mouse obesity induced by monosodium glutamate (administered to newborn mice) as it usually leads to increased CYP expression. Selecting an animal model that is as close as possible to the properties of human obesity is of paramount importance.

• MeSH
• Microsomes, Liver metabolism   MeSH
• Disease Models, Animal MeSH
• Mice MeSH
• Obesity physiopathology   MeSH
• Cytochrome P-450 Enzyme System metabolism   MeSH
• Adipose Tissue metabolism   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH