<p>OBJECTIVETo observe the effects of ginkgolides on gene expression of hepatic cytochrome P-450 in rats.p><p>METHODSprague-Dawley rats were administered ginkgolides (100 mg x kg(-1) body weight) through oral gavage once daily for four consecutive days. The level of gene expression in liver tissues was analyzed by competitive reverse transcription-polymerase chain reaction (competitive RT-PCR).p><p>RESULTA single and prospective band of CYP1A1, CYP1A2, CYP2B1/B2, CYP2C11, CYP2E1, CYP4A1 and cyclophilin was observed after polymerase chain reaction (PCR) when the reactive system of reverse transcription (RT) had no target RNA, which confirmed the competitor had a specific capacity to bind to the CYP or cyclophilin primer. CYP1A1 mRNA was not dectectable in the livers of untreated control rats and ginkgolides-treated rats. The levels of CYP2C11 and CYP2E1 were not changed by ginkgolides treatment. In contrast, the levels of gene expression for CYP1A2 and CYP2B1/B2 were decreased, however, the levels of gene expression for CYP3A1 and CYP4A1 in ginkgolides group were distinctly increased compared with the control.p><p>CONCLUSIONA specific effect of ginkgolides on cytochrome P-450 gene expression was observed in this investigation. Ginkgolides had various effects on different cytochrome P-450 isoforms.p>
Animals ; Aryl Hydrocarbon Hydroxylases ; biosynthesis ; genetics ; Cytochrome P-450 CYP1A1 ; biosynthesis ; genetics ; Cytochrome P-450 CYP1A2 ; biosynthesis ; genetics ; Cytochrome P-450 CYP2B1 ; biosynthesis ; genetics ; Cytochrome P-450 CYP3A ; Cytochrome P-450 Enzyme System ; biosynthesis ; genetics ; Cytochrome P450 Family 4 ; Gene Expression Regulation ; Ginkgo biloba ; chemistry ; Ginkgolides ; isolation & purification ; pharmacology ; Liver ; metabolism ; Male ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Rotundine (1 micromol L(-1)) was incubated with a panel of rCYP enzymes (1A2, 2C9, 2C19, 2D6 and 3A4) in vitro. The remained parent drug in incubates was quantitatively analyzed by an Agilent LC-MS. CYP2C19, 3A4 and 2D6 were identified to be the isoenzymes involved in the metabolism of rotundine. The individual contributions of CYP2C19, 3A4 and 2D6 to the rotundine metabolism were assessed using the method of total normalized rate to be 31.46%, 60.37% and 8.17%, respectively. The metabolites of rotundine in incubates were screened with ESI-MS at selected ion mode, and were further identified using MS2 spectra and precise molecular mass obtained from an Agilent LC/Q-TOF-MSMS, as well as MS(n) spectra of LC-iTrap-MS(n). The predominant metabolic pathway of rotundine in rCYP incubates was O-demethylation. A total 5 metabolites were identified including 4 isomerides of mono demethylated rotundine and one di-demethylated metabolite. The results also showed that CYP2C19, 2D6 and 3A4 mediated O-demethylation of methoxyl groups at different positions of rotundine. Furthermore, the ESI-MS cleavage patterns of rotundine and its metabolites were explored by using LC/Q-TOF-MSMS and LC/iTrap-MS(n) techniques.
Analgesics, Non-Narcotic ; metabolism ; Aryl Hydrocarbon Hydroxylases ; metabolism ; Berberine Alkaloids ; metabolism ; Chromatography, Liquid ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 CYP2C19 ; Cytochrome P-450 CYP2C9 ; Cytochrome P-450 CYP2D6 ; metabolism ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Dopamine Antagonists ; metabolism ; Humans ; Isoenzymes ; metabolism ; Methylation ; Recombinant Proteins ; metabolism ; Spectrometry, Mass, Electrospray Ionization

<p>OBJECTIVETo evaluate the influence of individual genetic polymorphisms of metabolic enzymes on urinary styrene metabolites.p><p>METHODS58 workers occupationally exposed to styrene were divided into the high exposure group (≥ 100 mg/m³) and the low exposure group (< 100 mg/m³). The microfluidic chip technology was used to determine the SNPs of CYP2B6, CYP2D6 and GSTP1 and the influence of gene polymorphisms on the metabolism of styrene was statistically analyzed.p><p>RESULTSThe level of urine styrene metabolites level was influenced by genotypes of CYP2B6, CYP2D6 and GSTP1 [(280.28 +/- 100.60) mg/g Cr vs (183.48 +/- 127.52) mg/g Cr, (233.04 +/- 77.56) mg/g Cr vs (152.46 +/- 95.47) mg/g Cr, (32.88 +/- 7.14) mg/g Cr vs (24.47 +/- 5.59) mg/g Cr, P < 0.05)]. The metabolism of CYP2B6 G/G homozygotic genotype to styrene was more active than G/T heterozygotic genotype and T/T mutation genotype. The level of PHEMA in GSTP1 homozygotic genotype subjects was significantly higher than that in the group of homozygotic genotype [(32.07 +/- 7.32) mg/g Cr vs (25.59 +/- 6.95) mg/g Cr, P < 0.05)]. The influence of CYP2D6 genotypes on urinary metabolites was also observed in the same study [(56.36 +/- 109.72) mg/g Cr vs (177.13 +/- 116.21) mg/g Cr, (118.73 +/- 84.55) mg/g Cr vs (148.48 +/- 99.83) mg/g Cr, (18.29 +/- 13.50) mg/g Cr vs (19.95 +/- 13.30) mg/g Cr, P < 0.05)].p><p>CONCLUSIONGenotypes of CYP2B6, GSTP1 and CYP2D6 are related to susceptibility to the metabolism of styrene in human.p>
Adult ; Aryl Hydrocarbon Hydroxylases ; genetics ; Cytochrome P-450 CYP2B6 ; Cytochrome P-450 CYP2D6 ; genetics ; Genotype ; Glutathione S-Transferase pi ; genetics ; Humans ; Male ; Middle Aged ; Occupational Exposure ; adverse effects ; Oxidoreductases, N-Demethylating ; genetics ; Polymorphism, Genetic ; Styrene ; adverse effects ; pharmacokinetics ; urine ; Young Adult

The paper is to report the study of the effect of Shenfu injection on the enzyme activity of liver CYP450 and its mRNA level of rat liver. Microsome of rat liver was prepared after intravenous administration of Shenfu injection for 7 days. The enzyme activity was quantified by Cocktail method. Meanwhile, the mRNA expression of CYP1A2, CYP2B1/2, CYP2C11 and CYP3A1 in the liver was detected by RT-PCR. Shenfu injection obviously induced the enzyme activities of CYP2B and CYP2C. Meantime Shenfu injection decreased the enzyme activities of CYP1A2 and CYP3A. The mRNA levels of CYP2B and CYP2C were also induced in rats treated with Shenfu injection. But it obviously inhibited the mRNA level of CYP1A2 and CYP3A. Since the enzyme activity and mRNA level were obviously changed after administration, the potential effect of drug-drug interaction should be concerned.
Aconitum ; chemistry ; Animals ; Aryl Hydrocarbon Hydroxylases ; genetics ; metabolism ; Cytochrome P-450 CYP1A2 ; genetics ; metabolism ; Cytochrome P-450 CYP2B1 ; genetics ; metabolism ; Cytochrome P-450 CYP3A ; genetics ; metabolism ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Cytochrome P450 Family 2 ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Injections ; Male ; Microsomes, Liver ; enzymology ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Steroid 16-alpha-Hydroxylase ; genetics ; metabolism

Abstract: The activities of four CYP450 enzymes (CYP3A, 1A2, 2El and 2C) and the mRNA expression levels of CYP1A2, 2El, 2Cll and 3A1 in rat liver were determined after Wistar rats were orally administered with brucine (BR) at three dosage levels (3, 15 and 60 mg.kg-1 per day) and the high dose of BR combined with glycyrrhetinic acid (GA, 25 mg.kg-1 per day) or liquiritin (LQ, 20 mg.kg-1 per day) for 7 consecutive days. Compared with the control, brucine caused 24.5% and 34.6% decrease of CYP3A-associated testosterone 6beta-hydroxylation (6betaTesto-OH) and CYP2C-associated tolbutamide hydroxylation (Tol-OH), respectively, and 146.1% increase of CYP2El-associated para-nitrophenol hydroxylation (PNP-OH) at the high dose level. On the other hand, (BR+GA) caused 51.4% and 33.5% decrease, respectively, of CYP2El-associated PNP-OH and CYP1A2-associated ethoxyresorufin-O-de-ethylation (EROD) as compared with the high dose of BR group. Meanwhile, (BR+LQ) caused 41.1% decrease of CYP2El-associated PNP-OH and 37.7% increase of CYP2C-associated Tol-OH. The results indicated that the co-administration of BR with GA or LQ had effect on mRNA expression and activities of the CYP450 enzymes mentioned above to some extent, and the in vivo antagonism of LQ on BR-induced CYPs adverse effects and the in vivo inhibitory action of GA on CYP2E1 and 1A2 might play an important role in the detoxification of Radix Glycyrrhizae against Strychnos nux-vomica L.
Animals ; Aryl Hydrocarbon Hydroxylases ; genetics ; metabolism ; Cytochrome P-450 CYP1A1 ; metabolism ; Cytochrome P-450 CYP1A2 ; genetics ; metabolism ; Cytochrome P-450 CYP2E1 ; genetics ; metabolism ; Cytochrome P-450 CYP3A ; genetics ; metabolism ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Cytochrome P450 Family 2 ; Flavanones ; pharmacology ; Gene Expression Regulation, Enzymologic ; Glucosides ; pharmacology ; Glycyrrhetinic Acid ; pharmacology ; Hydroxylation ; Liver ; enzymology ; metabolism ; Male ; Nitrophenols ; metabolism ; Plants, Medicinal ; chemistry ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar ; Steroid 16-alpha-Hydroxylase ; genetics ; metabolism ; Steroid Hydroxylases ; metabolism ; Strychnine ; analogs & derivatives ; isolation & purification ; pharmacology ; Strychnos nux-vomica ; chemistry ; Tolbutamide ; metabolism

The fields of pharmacogenetics and pharmacogenomics have become increasingly promising regarding the clinical application of genetic data to aid in prevention of adverse reactions. Specific screening tests can predict which animals express modified proteins or genetic sequences responsible for adverse effects associated with a drug. Among the genetic variations that have been investigated in dogs, the multidrug resistance gene (MDR) is the best studied. However, other genes such as CYP1A2 and CYP2B11 control the protein syntheses involved in the metabolism of many drugs. In the present study, the MDR-1, CYP1A2 and CYP2B11 genes were examined to identify SNP polymorphisms associated with these genes in the following four canine breeds: Uruguayan Cimarron, Border Collie, Labrador Retriever and German Shepherd. The results revealed that several SNPs of the CYP1A2 and CYP2B11 genes are potential targets for drug sensitivity investigations.
Animals ; Aryl Hydrocarbon Hydroxylases/*genetics/metabolism ; Cytochrome P-450 CYP1A2/*genetics/metabolism ; Dogs/*genetics/metabolism ; P-Glycoprotein/*genetics/metabolism ; *Polymorphism, Single Nucleotide ; Steroid Hydroxylases/*genetics/metabolism

<p>AIMTo identify the drug-metabolizing enzymes involved in the hydroxylation of the new anti-inflammatory and anodyne imrecoxib.p><p>METHODSImrecoxib was incubated with heterologous expression human cytochrome P450 (rCYPs) in vitro, and metabolites and remained parent drug were detected with liquid chromatography-multistage mass spectrometry. The contribution of 4 CYPs in the hydroxylation metabolism of imrecoxib was evaluated by total normalized rate (TNR) method.p><p>RESULTSImrecoxib is metabolized by CYP2C9, CYP2D6 and CYP3A4, with the rate of 62.5%, 21.1% and 16.4%, respectively.p><p>CONCLUSIONCYP2C9 is the major enzyme involved in imrecoxib hydroxylation metabolism.p>
Aryl Hydrocarbon Hydroxylases ; metabolism ; Cyclooxygenase 2 Inhibitors ; metabolism ; Cytochrome P-450 CYP2C9 ; Cytochrome P-450 CYP2D6 ; metabolism ; Cytochrome P-450 CYP3A ; Cytochrome P-450 Enzyme System ; metabolism ; Hydroxylation ; Pyrroles ; metabolism ; Spectrometry, Mass, Electrospray Ionization ; Sulfides ; metabolism

<p>OBJECTIVETo detect the effect of deltamethrin (DM) on benzyloxyresorufin O-dealkylatase (BROD) activity and the expression of CYP2B1/2B2 in rat brain.p><p>METHODSBROD activity was determined by fluorophotometry under the treatment of DM in vivo and vitro. Western-blot analysis was used to detect the expression of CYP2B1/2B2.p><p>RESULTSIn vivo, DM could markedly inhibit BROD activity in rat brain microsome after rats had been treated with DM (12.5 mg.kg-1.d-1, i.p.) for 5 days. The inhibitory rate in whole brain, cerebral cortex and cerebellum were 26.7%, 23.8% and 33.3% respectively(P < 0.05). However, in vitro, under the concentration of 2 x 10(-8)-2 x 10(-4) mol/L of DM, there was no obvious change of BROD activity in rat brain. Moreover, Western-blot analysis indicated that DM could significantly reduce the expression of CYP2B1/2B2 in vivo, the inhibitory rate of protein synthesis was 42.6%.p><p>CONCLUSIONDM could inhibit BROD activity in rat brain and this effect may be related to the reduction of CYP2B1/2B2 protein synthesis.p>
Animals ; Aryl Hydrocarbon Hydroxylases ; antagonists & inhibitors ; biosynthesis ; Blotting, Western ; Brain ; drug effects ; enzymology ; Cytochrome P-450 CYP2B1 ; antagonists & inhibitors ; biosynthesis ; Enzyme Inhibitors ; toxicity ; Insecticides ; toxicity ; Nitriles ; toxicity ; Pyrethrins ; toxicity ; Rats ; Steroid Hydroxylases ; antagonists & inhibitors ; biosynthesis

<p>OBJECTIVETo examine the expression of cytochrome P450 related genes in oral submucous fibrosis tissue and to investigate the possible role of the genes in pathogenesis of oral submucous fibrosis (OSF).p><p>METHODSBuccul mucosa tissues were obtained from OSF patients in early, medium and advanced stages, with each stage including 10 patients. Normal buccul mucosa tissues were collected from 10 patients undergoing oral and maxillofacial surgery as control. Oral submucous fibrosis-related genes were analysed by cDNA chips, and the results were submitted to the gene network database. Differentially expressed genes related to the pathway of CYP metabolism were indentifyed by the database analysis. Reverse transcription-polymerase chain reaction (RT-PCR) was used to verify the results from cDNA chips by increasing sample volume.p><p>RESULTSThere were eight genes [CYP2B6, CYP2C18, CYP2F1, CYP3A5, microsomal glutathione S-transferase 2 (MGST2), alcohol dehydrogenase (ADH), UDP glucuronosyl transferase 2B15 (UGT2B15), ADH1C] which were related to the pathway of CYP metabolism. These genes were low expressed in all stages of OSF (P < 0.001).There were no differences in genes expression among the three stages of OSF (P > 0.05).p><p>CONCLUSIONSThere were down-regulated genes related to the pathway of CYP metabolism in oral submucous fibrosis tissue. The ability of the pathway of CYP to metabolize and clear betel nut ingredients was reduced in OSF patients, which may play a role in the pathogenesis of OSF.p>
Adult ; Alcohol Dehydrogenase ; genetics ; metabolism ; Aryl Hydrocarbon Hydroxylases ; genetics ; metabolism ; Cytochrome P-450 CYP2B6 ; Cytochrome P-450 CYP3A ; genetics ; metabolism ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Cytochrome P450 Family 2 ; Down-Regulation ; Glucuronosyltransferase ; genetics ; metabolism ; Glutathione Transferase ; genetics ; metabolism ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; Oral Submucous Fibrosis ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult

<p>AIMTo evaluate the effect of in vitro and in vivo treatment with mitomycin C (MMC) on activities of CYP2D1/2, CYP2C1 , and CYP1A2 in the liver of male rats.p><p>METHODSUsing HPLC to determine the activities of the three isoenzymes in rat liver microsomes by detecting the specific metabolites of their substrates after treatment with inducers in vivo or inhibitors in vitro.p><p>RESULTSIn vitro, MMC inhibited the activity of CYP2D1/2, CYP2C11, and CYP1A2 in dexamethasone-induced microsomes by (19 +/- 6)% (P < 0.05), (85 +/- 10)% (P < 0.01), and (36 +/- 6)% (P < 0.05), respectively, and decreased the activity of CYP1A2 in beta-naphthoflavone-induced microsomes by (58 +/- 6)% (P < 0.01). Rats were injected intraperitoneally with 20% of the LD50 of MMC for 3 or 6 d. The treatment showed no significant effect on microsomal activities of CYP2D1/2, CYP2C11 or CYP1A2.p><p>CONCLUSIONMMC can inhibit the activities of CYP2D1/2, CYP2C11, and CYP1A2 in rat liver microsomes in vitro, but it showed no significant effect on the activities of the three isoenzymes in vivo.p>
Alcohol Oxidoreductases ; Animals ; Antibiotics, Antineoplastic ; pharmacology ; Aryl Hydrocarbon Hydroxylases ; metabolism ; Biotransformation ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P450 Family 2 ; Male ; Microsomes, Liver ; enzymology ; Mitomycin ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Steroid 16-alpha-Hydroxylase ; metabolism