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1.
Effect of a single oral dose of methanol, ethanol and propan-2-ol on the hepatic microsomal metabolism of foreign compounds in the rat.
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G Powis 《The Biochemical journal》1975,148(2):269-277
Methanol and ethanol administered to rats as a single oral dose increased aniline hydroxylation by the hepatic microsomal fraction by a maximum of 169 and 66% respectively, whereas aminopyrine demethylation was inhibited by 51 and 61%. The concentration of microsomal cytochrome P-450, and the activities of NADPH-cytochrome c reductase and NADPH-cytochrome P-450 reductase were unchanged. Propan-2-ol, administered as a single oral dose, increased microsomal aniline hydroxylation by 165% and increased aminopyrine demethylation by 83%. The concentration of cytochrome P-450 was unchanged whereas NADPH-cytochrome c reductase and NADPH-cytochrome P-450 reductase were both increased by 38%. Methanol, ethanol and propan-2-ol administration resulted in a decreased type I spectral change but had no effect on the reverse type I spectral change. Methanol administration decreased the type II spectral change whereas ethanol and propan-2-ol had no effect. Cycloheximide blocked the increases in aniline hydroxylation and aminopyrine demethylation but could not completely prevent the decreases in aminopyrine demethylation. The increases in aniline hydroxylation were due to an increase in V, but Km was unchanged. The ability of acetone to enhance and compound SKF 525A to inhibit microsomal aniline hydroxylation was decreased by the administration of all three alcohols. The decrease in the metabolism of aminopyrine may result from a decrease in the binding to the type I site with a consequent failure of aminopyrine to stimulate the reduction of cytochrome P-450. Methanol administration may lead to an increase in aniline hydroxylation because of a failure of aniline to inhibit cytochrome P-450 reduction. 相似文献
2.
The in vitro metabolism of p-nitroanisole, aminopyrine, and aniline by rat liver microsomal monoxygenases were studied in the presence of different polychlorinated biphenyl (PCB) mixtures and some related hydroxybiphenyls. The tested PCB mixtures contained preferably dichloro- (di-CB), tetrachloro- (tetra-CB), or hexachlorobiphenyls (hexa-CB). All PCB were competitive inhibitors of only aminopyrine demethylation by normal microsomes (Ki 22-39 micron). In microsomes of PCB-pretreated rats the aminopyrine demethylation was inhibited noncompetitively by di-CB and hexa-CB whereas tetra-CB remained a competitive inhibitor (Ki 12 micron). Moreover, after PCB pretreatment all PCB were competitive inhibitors of p-nitroanisole demethylation. 2-OH-biphenyl and 4-OH-biphenyl caused competitive inhibition of aminopyrine demethylation and aniline hydroxylation but failed to inhibit p-nitroanisole metabolism by normal microsomes. Chlorinated 4-hydroxybiphenyls inhibited competitively the metabolism of both type I and type II substrates. However, after PCB pretreatment all phenolic compounds caused uncompetitive inhibition of aniline hydroxylation. 相似文献
3.
The role of hydrophobicity and electronic factors in regulating alcohol inhibition of cytochrome P-450-mediated aniline p-hydroxylation has been investigated by the formulation of quantitative structure-activity relationships. The activity of linear primary alcohols and unhindered linear secondary alcohols shows a linear dependence on log P, where P is the octanol-water partition coefficient. Hindered primary and secondary alcohols are less active than this relationship predicts. An equation describing the activity of both hindered and unhindered primary and secondary alcohols shows that alcohol inhibition of aniline hydroxylation is regulated by hydrophobicity and steric effects. No role for electronic factors can be discerned. Similarities are found between alcohol inhibition and the binding of alkyl amines to cytochrome P-450, suggesting that alcohols may bind to the amine binding site. 相似文献
4.
Magnus Ingelman-Sundberg Gunilla Ekström 《Biochemical and biophysical research communications》1982,106(2):625-631
Hydroxylation of aniline, catalyzed by rabbit liver microsomal cytochromes P-450 in reconstituted systems, was inhibited by catalase, superoxide dismutase, catechol, mannitol, hydroquinone, dimethylsulfoxide and benzoate, whereas the cytochrome P-450-catalyzed O-demethylation of paranitroanisole, measured under the same conditions, was unaffected by these agents. A similar inhibition profile of the hydroxylation reaction was observed in reconstituted systems where cytochrome P-450 had been replaced by hemoglobin. The results indicate that aniline hydroxylation is mediated by hydroxyl radicals generated in an iron-catalyzed Haber-Weiss reaction between O2? and H2O2 and may explain some of the special properties of this reaction previously described. 相似文献
5.
Acetone was found to enhance both cumene hydroperoxide and NADPH/oxygen supported microsomal aniline hydroxylation. Since cumene hydroperoxide supported aniline hydroxylation does not involve an electron transport system, these results suggest that acetone produces its enhancing effect by facilitating either the formation of an activated oxygen species or the insertion of oxygen into the substrate. 相似文献
6.
The effect of riboflavin deficiency and phenobarbital pretreatment on drug hydroxylation and lipid peroxidation was investigated. A significant decrease in aniline and acetanilide hydroxylation as well as NADPH-linked and ascorbate-induced lipid peroxidation was observed during 4- and 7-week riboflavin deficiency in both adult male and adult female rats. The drug-hydroxylation and lipid-peroxidation activities were further lowered with the increase in riboflavin deficiency. The phenobarbital pretreatment induced aniline and acetanilide hydroxylase activity even in riboflavin-deficient animals. Drug hydroxylation inhibits lipid peroxidation in both deficient and normal rats. The administration of riboflavin was followed by a significant increase in drug hydroxylation and lipid peroxidation. 相似文献
7.
Changes in the ultraviolet/visible spectrum of human oxyferrohemoglobin upon addition of aniline were indicative of a concentration-dependent interaction of aniline with hemoglobin, resulting in accelerated autooxidation of the hemoprotein. Oxygen was found to markedly inhibit this interaction of aniline with oxyhemoglobin. The dependence of the rate of autooxidation on aniline concentration followed saturation kinetics and showed a half-maximal response at 8 mM aniline. This value is equal to the value of Km for aniline as substrate for the O2-dependent, hemoglobin-catalyzed hydroxylation reaction which yields p-aminophenol (Mieyal, J. J., Ackerman, R.S., Blumer, J.L., and Freeman, L.S. (1976) J. Biol. Chem. 241, 3436-3441). Thus, an aniline-oxyhemoglobin complex is implicated in the overall catalytic reaction. No detectable p-aminophenol was formed when aniline was combined with oxyhemoglobin in the absence of an electron donor, but hydroxylation of aniline does occur when NADPH, NADPH plus P-450 reductase, or Na2S2O4 are also added. 相似文献
8.
M Osada Y Ogura H Yasui H Sakurai 《Biochemical and biophysical research communications》1999,263(2):392-397
Cytochrome P450 (P450)-dependent p-hydroxylation of aniline and o-deethylation of 7-ethoxycoumarin were examined in rat liver microsomes in the presence of radical scavengers. The addition of beta-carotene, a quencher of singlet oxygen species ((1)O(2)), suppressed the aniline hydroxylation, while the addition of sodium azide (NaN(3)) ((1)O(2) quencher) enhanced the reaction. No other reactive oxygen scavengers or chelating agents such as superoxide dismutase, catalase, dimethylsulfoxide, or deferoxamine altered the reaction. In contrast, the microsomal o-deethylation of 7-ethoxycoumarin was suppressed by the addition of NaN(3). (1)O(2) was detectable during the reaction of microsomes and NADPH by ESR spin-trapping when 2,2,6,6-tetramethyl-4-piperidone (TMPD) was used as a spin trap, and the (1)O(2) was quenched by the additions of beta-carotene, NaN(3), aniline, and 7-ethoxycoumarin. The enhancement effect of NaN(3) in the hydroxylation of aniline appeared to be due to the conformational change of P450 protein, which in turn enhances the binding of aniline to P450 in terms of the spectral dissociation constant (K(s)). In contrast, (1)O(2) appeared to be active in the o-deethylation of 7-ethoxycoumarin. On the basis of the results, the involvement of (1)O(2) in P450-dependent substrate oxygenations is proposed. 相似文献
9.
P F Han G Y Han H C McBay J Johnson 《Biochemical and biophysical research communications》1978,85(2):747-755
A number of agents were tested for their ability to enhance the p-hydroxylation of aniline using isolated hepatocytes as a model system. Although the observed stimulation or inhibition was not concentration dependent, various substrates for the hepatic mixed-function oxygenase (MFO) system (p-nitroanisole, 7-ethoxycoumarin, biphenyl, N,N′-dimethylaminoazobenzene, and benzphetamine) stimulated the hydroxylation at a concentration of 0.5 mm. This effect was not seen with all substrates. In general, aniline hydroxylation was not affected by the other agents tested (steroids, metabolic inhibitors and MFO inhibitors). However, enhancement was noticed with testosterone and progesterone at the lowest concentration (0.05 mm), with 2,6-dichloro-4-nitrophenol and salicylamide at 0.05 mm and 0.5 mm and with 7,8-benzoflavone at 5.0 mm. 相似文献
10.
Levels of protein kinase C and nitric oxide synthase activity in rats exposed to sub chronic low level lead 总被引:1,自引:0,他引:1
Previous studies from our laboratories have linked the protective abilities of IH636 grape seed proanthocyanidin extract (GSPE) with inactivation of anti-apoptotic gene bcl-XL, and modification of several other critical molecular targets such as DNA-damage/DNA-repair, lipid peroxidation and intracellular Ca2+ homeostasis. Especially, GSPE provided dramatic protection against acetaminophen (APAP)-induced hepatotoxicity, significantly increased bcl-XL expression in the liver [1], and antagonized both necrotic and apoptotic deaths of liver cells in vivo. However, it was not clear from this study whether anti-apoptogenic and anti-necrotic effects of GSPE were: (i) due to its interference with endonuclease activity, (ii) due to its antioxidant effect, or, (iii) due to its ability to inhibit microsomal drug metabolizing enzyme(s), such as CYP-4502E1. Since CYP-4502E1 primarily metabolizes acetaminophen in mice and rats, this study specifically focused on CYP-4502E1's catalytic activity in vitro. Overall this investigation compared the in vitro aniline hydroxylation patterns of: (i) in vivo GSPE-exposed and unexposed (control) mouse liver microsomes, (ii) induced (1% acetone in drinking water for 3 days) and uninduced rat liver microsomes in the presence and absence of GSPE in vitro, and (iii) control rat liver microsomes in the presence of an anti-APAP agent 4-aminobenzamide (4-AB) in vitro. For the in vivo assessment, male B6C3F1 mice were fed GSPE diet (ADI 100 mg/kg body wt) for 4 weeks, and liver microsomes were isolated from both control and GSPE-fed mice for aniline hydroxylation, a specific marker of CYP-4502E1 activity. Data show that hydroxylation was 40% less in microsomes from GSPE-exposed livers compared to control microsomes. Similarly, when rat liver microsomes were incubated with various concentrations of GSPE in vitro (100 and 250 g/ml), aniline hydroxylation was inhibited to various degrees (uninduced: 40 and 60% and induced: 25 and 50%, respectively with 100 and 250 g/ml). Influence of GSPE on hydroxylation patterns were compared with another hepatoprotective agent 4-aminobenzamide (4-AB), a well-known modulator of nuclear enzyme poly(ADP-ribose) polymerase, and the data shows that 4-AB did not alter aniline hydroxylation at all. Collectively, these results may suggest that GSPE has the ability to inhibit CYP-4502E1, and this is an additional cytoprotective attribute, in conjunction with its novel antioxidant and/or antiendonucleolytic potential. 相似文献
11.
D F Lewis 《Chemico-biological interactions》1987,62(3):271-280
The results of Molecular Orbital (MO) calculations by the MINDO/3 method are reported, together with the results of multiple regression analysis of electronic and structural parameters with inhibition of aniline hydroxylation by a series of 22 alcohols. The most significant correlations show the relationships between molecular length, frontier electron density of the alpha-carbon and hydroxyl oxygen, nucleophilic superdelocalizability of the hydroxyl hydrogen, energy of the highest occupied MO and biological activity involving binding to microsomal cytochromes P-450. Using the data of Cohen and Mannering (Mol. Pharmacol., 9 (1973) 383), Testa, (Chem.-Biol. Interact., 34 (1981) 287) has shown that the inhibition of aniline hydroxylation by a series of alcohols can be related to their electronic structure and hydrophobicity (measured by log P, the octanol-water partition coefficient). The mode of binding and effect on spin-state equilibria in cytochrome P-450 by alcohols has been elucidated by Testa, whereas an alternative hypothesis based on connectivity correlations has been reported by Sabljic and Sabljic (Mol. Pharmacol., 23 (1983) 213). The present work shows that the biological response can be explained by calculated electronic structure and molecular shape parameters. Also, one compound (the only tertiary alcohol) from the original set that was not included in Testa's calculations and analysis, is included in this work and its activity successfully calculated. The latter authors, Sabljic and Sabljic, were led to exclude the data for this compound and one other (phenyl methanol) in order to achieve a good correlation with their calculated parameters of molecular structure. 相似文献
12.
Benzphetamine demethylase and aniline hydroxylase activities were determined with various hemoproteins including indoleamine 2,3-dioxygenase in a cytochrome P-450-like reconstituted system containing NADPH, NADPH-cytochrome P-450 reductase, and O2. The highest specific activities, almost comparable to those of liver microsomal cytochrome P-450, were detected with indoleamine 2,3-dioxygenase from the rabbit intestine. The indoleamine 2,3-dioxygenase-catalyzed benzphetamine demethylation reaction was inhibited by catalase but not by superoxide dismutase. Exogenous H2O2 or organic hydroperoxides was able to replace the reducing system and O2. The stoichiometry of H2O2 added to the product formed was essentially unity. These results indicate that the dioxygenase catalyzes the demethylation reaction by the so-called "peroxygenation" mechanism using H2O2 generated in the reconstituted system. On the other hand, the dioxygenase-catalyzed aniline hydroxylation reaction was not only completely inhibited by catalase but also suppressed by superoxide dismutase by about 60%. Although the O2- and H2O2-generating system (e.g. hypoxanthine-xanthine oxidase) was also active as the reducing system, neither exogenous H2O2 nor the generation of O2- in the presence of catalase supported the hydroxylation reaction, indicating that both H2O2 and O2- were essential for the hydroxylation reaction. However, typical scavengers for hydroxyl radical and singlet oxygen were not inhibitory. These results suggest that a unique, as yet unidentified active oxygen species generated by H2O2 and O2- participates in the dioxygenase-mediated aniline hydroxylation reaction. 相似文献
13.
I I Karuzina G I Bachmanova G P Kuznetsova M V Izotov A I Archakov 《Biokhimii?a (Moscow, Russia)》1979,44(10):1796-1804
It has been found that NADPH-dependent hydroxylation of dimethylaniline, aniline, p- and o-nitroanisol and lipid peroxidation is inhibited by the tyrosine-copper (II) complex (low molecular weight analog of superoxide dismutase), which is indicative of a possibility of superoxide radicals formation in these reactions. The inhibition of the above-mentioned reactions with Tyr2-Cu2+ is less pronounced or absent, if cumole hydroperoxide is used as cosubstrate instead of NADPH. Differences in the Tyr2-Cu2+ complex effects on the cumule hydroperoxide-dependent xenobiotics hydroxylation and lipid peroxidation catalyzed by various forms of cytochrome P-450, e. g. microsomal, soluble and incorporated into liposomes, have been found. The data obtained suggest that the efficiency of the inhibitory effect of the Tyr2-Cu2+ complex depends on the type of cosubstrates (NADPH, cumole hydroperoxide) and substrates used as well as on the form of cytochrome P-450. 相似文献
14.
The role of hydroxylation in the secretion of collagen by mouse fibroblasts in culture 总被引:3,自引:0,他引:3
The secretion of collagen may be specifically and quantitatively measured in a cell culture system using mouse fibroblasts, line 3T6. The results of such a study show that inhibition of the hydroxylation of collagen with α,α′-dipyridyl reduces the rate of secretion to 27% of the rate of normally hydroxylated collagen. Dipyridyl does not hinder passage of the normally hydroxylated collagen species to the medium. Therefore its effect on the secretion of under-hydroxylated collagen (protocollagen) is due to its inhibition of hydroxylation and not to an inhibition of the cell's secretory machinery. 相似文献
15.
Changes in the activity of a NADPH-dependent monooxygenase system of the rat liver are studied under the effect of tetramethylthiuramdisulphide. Under these conditions aniline hydroxylation is shown to be inhibited to a higher extent than amidopyrine demethylation. Besides a decrease in the level of cytochrome P-450, the central component of the microsomal system of hydroxylation, there appears cytochrome P-420--an inactivated form of cytochrome P-450. 相似文献
16.
An electrochemical system of cytochrome P-450 reduction in the presence of the water-soluble redox carrier methylviologen has been developed. In this system cytochrome P-450 effectuates a steady-state demethylation of dimethylaniline and hydroxylation of aniline. The results of control experiments suggest that the above reactions are mediated by cytochrome P-450. The effect of the peroxidase reaction is excluded by an addition of high concentrations of catalase to the incubation mixture. At the same time the hydroxylation of these substrates is accompanied by methylviologen demethylation. 相似文献
17.
V Iu Uvarov A V Leshchenko I G Rukavishnikov Ch S Dzhuzenova A I Archakov 《Biokhimii?a (Moscow, Russia)》1988,53(9):1433-1438
The effect of intramolecular cross-links formation in isolated cytochrome P-450 LM2 on its reactivation after incorporation into the liposome lipid bilayer was studied. Treatment with bifunctional reagents results in the inactivation of the solubilized haemoprotein. The degree of the enzyme immobilization determines the degree of inhibition of p-nitroanisol demethylation and aniline hydroxylation. Whereas the complete inhibition of oxidation of type II substrate turnover needs two intramolecular cross-links, that of type I substrates necessitates at least seven cross-links. The incorporation of modified and native enzymes into the membrane lipid bilayer at temperatures above the phase transition point results in the enzyme activation. However, in case of the preimmobilized enzyme the activation does not reach the maximal values. Both stabilized and liposome-incorporated cytochrome P-450 can fully be reactivated via the cross-link disulfide bond reduction. No such effect is observed at temperatures below the phase transition point. 相似文献
18.
Henrik Anttinen Ulla Puistola Taina Pihlajaniemi Kari I. Kivirikko 《Biochimica et Biophysica Acta (BBA)/General Subjects》1981,674(3):336-344
The addition of Zn2+ inhibited lysine hydroxylation markedly less effectively than it did proline hydroxylation in chick embryo tendon cells, 3T6 fibroblasts and lysyl hydroxylase-deficient Ehlers-Danlos Syndrome Type VI fibroblasts. With low Zn2+ concentrations, a similar difference was also seen in chick embryo cartilage cells, whereas with high concentrations both hydroxylations were affected to the same extent in this cell type. Ascorbate deficiency likewise had a much less effect on lysine than proline hydroxylation when studied with 3T6 fibroblasts. As these two effectors involve quite different mechanisms, it is suggested that relative insensitivity to inhibition may be a property of lysine hydroxylation seen in many cell types with a number of agents.Studies on the mechanism of the difference in the inhibition indicates that the phenomenon is probably not due to differences in the kinetic constants of Zn2+ and ascorbate for the two enzymes. Neither is it probably to any major extent due to delayed procollagen triple helix formation nor a difference in the location of the two hydroxylases within the cisternae of the rough endoplasmic reticulum. The difference similarly cannot be explained solely by an excess of lysyl hydroxylase in the cell. It may thus be due either to some other intrracellular property or to the combined effect of several factors. 相似文献
19.
M E Letelier E Del Villar E Sanchez 《Comp. Biochem. Physiol. C, Comp. Pharmacol. Toxicol.》1985,80(1):195-198
Octodon degus shows greater tolerance to pentobarbital as compared with the Wistar rat. Mixed function oxidase activities in liver microsomes were higher in Octodon degus than in the Wistar rat. The reactions assayed were: aminopyrine N-demethylation, aniline and naphthalene hydroxylation and p-nitroanisole O-demethylation. These higher activities seem to be due mainly to the greater cytochrome P-450 content of liver microsomes of Octodon degus. Glutathione S-transferase activity towards 1-chloro-2,4-dinitrobenzene was 30 times higher in Octodon degus than in the Wistar rat. These results may explain the tolerance of Octodon degus to pentobarbital and other drugs. 相似文献
20.
The ability of trout to metabolize aniline in vitro in the presence of some divalent metal ions was investigated in the liver microsomes of rainbow trout, Salmo gairdneri. Trout liver microsomes were highly capable of catalyzing aniline hydroxylation to p-aminophenol with a specific activity of 0.068 nmoles/min per mg of microsomal protein in potassium phosphate buffer, pH 7.4 at 25°C. The activity of the aniline hydroxylase system was competitively inhibited by Hg+2, Ni+2, Cd+2, and Zn+2, while Cu+2 and Fe+3 seemed to inhibit the activity noncompetitively at 1 mM aniline concentrations. IC50 values at fixed aniline concentration were estimated to be 0.45 mM for Hg+2, Ni+2, and Cd+2, 1.8 mM for Zn+2 and Fe+3, and 1.3 mM for Cu+2. Eadie-Hofstee plots gave identical Vmax values of approximately 0.046 nmol/min per mg of protein while Km values were increased in the presence of Hg+2, Ni+2, CD+2, and Zn+2, indicating competitive inhibition. Both Km and Vmax values were affected by Fe+3 and Cu+2, suggesting noncompetitive inhibition. Ki values extracted from the Dixon plots were determined t be 0.23, 0.43, and 0.65 mM for Hg+2, Ni+2, and Cd+2, respectively, providing the most effective inhibition on the aniline hydroxylase system among studied metal ions. The Ki values were much higher in the presence of others. The results indicate a selective inhibition of the aniline hydroxylase system of trout liver microsomes by divalent metal ions. © 1997 John Wiley & Sons, Inc. 相似文献