The present work was to study induction of cytochrome P450 (CYP)3A and CYP2H1 gene by reverse transcriptase polymerase chain reaction (RT-PCR) and quantitative RTPCR in Bantam, Bantamized White Leghorn and White Leghorn chicks. Out of 18 chicks total 3 from each group (Bantam, Bantamized White Leghorn and White Leghorn) were treated intraperitoneal with phenobarbital at the dose rate of 12 mg/100 g (body weight) while the control group was treated with the saline. Total RNA was extracted from the liver samples using Tri Reagent based method. First strand cDNA was synthesized using one step RT-PCR kit. The PCR was performed and the product was subjected to agarose gel electrophoresis. Quantitative RT-PCR was conducted to quantify gene expression level of CYP3A and CYP2H1 genes. Relative expression ratio of CYP3A and CYP2H1 genes was calculated using relative expression software tool (REST). It was found that CYP3A is up regulated by factor of 1.34, 14.51 and 1.00 in Bantam, Bantamized White Leghorn and White Leghorn chicks, respectively. In Bantam and Bantamized White Leghorn chicks CYP2H1 gene was up regulated by factor 1.50 and 80.87, respectively but down regulated by a factor of 1.97 in White Leghorn chicks. The PCR efficiency ranged from 1.30 to 1.70, 0.86 to 1.70 and 0.91 to 1.58 for CYP3A, CYP2H1 and beta-actin, respectively in Bantam, Bantamized White Leghorn and White Leghorn chicks.
Animals ; Chickens/*metabolism ; Cytochrome P-450 CYP3A/*biosynthesis/genetics ; Cytochrome P-450 Enzyme System/*biosynthesis/genetics ; Gene Expression Regulation/drug effects ; Phenobarbital/*pharmacology ; Reverse Transcriptase Polymerase Chain Reaction

<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

<p>OBJECTIVETo investigate whether kaempferol stimulates pregnane X receptor (PXR)-mediated transcription of CYP3A4.p><p>METHODSTransient cotransfection reporter gene assay was performed with PXR expression plasmid and a reporter plasmid containing the XREs in the CYP3A4 gene promoter in HepG(2)cells.p><p>RESULTSKaempferol activated PXR-mediated transcription of CYP3A4 in a dose, time-dependent manner. In the dose-response study, kaempferol exposure at concentrations of 1.0 x 10(-3), 1.0 x 10(-2), 0.1, 1.0 and 10.0 mol/L for 24 h increased CYP3A4 transcription by (1.31+/-0.27), (1.45+/-0.36), (1.96+/-0.50), (2.90+/-1.07) and (7.93+/-0.75) fold, respectively compared with 0.1% DMSO (P<0.05). The results from time-course study showed that after 48 h exposure 1.0 and 10.0 mol/L of kaempferol enhanced the transcription of CYP3A4 by (3.73+/-1.21) fold and (8.42+/-1.47) fold, respectively.p><p>CONCLUSIONKaempferol may be a human CYP3A4 gene inducer through PXR, and may affect the metabolism of a large number of substrates of CYP3A4 simultaneously taken.p>
Carcinoma, Hepatocellular ; pathology ; Cytochrome P-450 CYP3A ; Cytochrome P-450 Enzyme System ; biosynthesis ; genetics ; Dose-Response Relationship, Drug ; Genes, Reporter ; Humans ; Kaempferols ; pharmacology ; Liver Neoplasms ; pathology ; Receptors, Steroid ; metabolism ; Transcription, Genetic ; Transfection ; Tumor Cells, Cultured

<p>OBJECTIVETo test the effect on human pregnane X receptor (hPXR)-mediated transcription regulation of CYP3A4 by five selected phytochemicals.p><p>METHODSTransient cotransfection reporter gene assays in HepG(2) cells were performed with the hPXR expression plasmid and the reporter gene plasmid which contains XRE in the promoter of CYP3A4 linked to luciferase.p><p>RESULTSIn the dose-effect study, soybean isoflavone, luteolin and curcumin induced the CYP3A4 transcription via PXR in an evident dose-dependent manner, but isorhamnetin and rutin did not. The inducibility of soybean isoflavone, luteolin and curcumin was also increased in concentrations between 1 micromol/L and 50 micromol/L, 24 h after induction, 50 micromol/L soybean isoflavone, luteolin and curcumin exhibited a 5.46-fold, 2.87-fold, and 2.07-fold increase respectively, compared with 0.1% DMSO treated cells. In the time-effect study, 10 micromol/L and 50 micromol/L soybean isoflavone, luteolin and curcumin induced CYP3A4 transcription between 12 h and 48 h, the strongest induction appeared in 48 h. 48 h after induction, 50 micromol/L soybean isoflavone, luteolin and curcumin exhibited a 6.72-fold, 3.24-fold, and 2.13-fold increase respectively, compared with 0.1% DMSO treated cells.p><p>CONCLUSIONThree phytochemicals, i.e. soybean isoflavone, luteolin and curcumin stimulate the PXR-mediated transcription of CYP3A4. Isorhamnetin and rutin have no effect on the CYP3A4 transcription via PXR.p>
Carcinoma, Hepatocellular ; pathology ; Curcumin ; pharmacology ; Cytochrome P-450 CYP3A ; Cytochrome P-450 Enzyme System ; biosynthesis ; genetics ; Humans ; Isoflavones ; pharmacology ; Liver Neoplasms ; pathology ; Luteolin ; pharmacology ; Plant Preparations ; pharmacology ; Receptors, Steroid ; metabolism ; Soybeans ; chemistry ; Transcription, Genetic ; Transfection ; Tumor Cells, Cultured

<p>OBJECTIVETo study whether zearalenone (ZEA), a fungal estrogen, can transcriptionally up-regulate the expression of cytochrome 450 3A4 (CYP3A4) transcription by activating human steroid hormone and xenobiotic receptor (SXR).p><p>METHODTransient cotransfection reporter gene assays were performed with human SXR expression plasmid and a reporter plasmid containing the SXR in the CYP3A4 gene promoter in HepG(2) cells.p><p>RESULTSThe transcriptional induction of CYP3A4 by ZEA with a dose, time-dependent manner. ZEA at the concentrations of 0.01, 0.10, 1.00 and 10.00 micromol/L, respectively, could induce CYP3A4 with (1.50 +/- 0.21), (1.66 +/- 0.27), (3.04 +/- 0.82) and (3.96 +/- 1.16) folds, as compared with 0.1% DMSO. Results from a time-dependent study show that 1.00 and 10.00 micromol/L of ZEA for 12 to 48 hours could enhance the transcription of CYP3A4 with (3.69 +/- 1.34) and (5.18 +/- 1.50) folds, and 10.00 micromol/L of ZEA for 48 hours could induce the CYP3A4 gene expression (5.18 +/- 1.50) folds, as compared with 0.1% DMSO by activating human SXR.p><p>CONCLUSIONZEA could induce the expression of the CYP3A4 gene transcription through activating SXR, possibly by affecting the other substrates of the CYP3A4, especially affecting the metabolism of drugs in the body.p>
Carcinoma, Hepatocellular ; pathology ; Cytochrome P-450 CYP3A ; Cytochrome P-450 Enzyme System ; biosynthesis ; genetics ; Genes, Reporter ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Transcription, Genetic ; drug effects ; Tumor Cells, Cultured ; Zearalenone ; pharmacology

Animals ; Bridged Bicyclo Compounds ; metabolism ; Cytochrome P-450 CYP3A ; biosynthesis ; genetics ; Drug Design ; Drug Interactions ; Enzyme Induction ; Humans ; Lithocholic Acid ; metabolism ; Phloroglucinol ; analogs & derivatives ; metabolism ; Receptors, Cytoplasmic and Nuclear ; genetics ; metabolism ; physiology ; Receptors, Steroid ; genetics ; physiology ; Signal Transduction ; Terpenes ; metabolism ; Transcription Factors ; genetics ; metabolism

<p>OBJECTIVETo study the effects of 18beta-glycyrrhizic acid and 18alpha-glycyrrhizic acid on mRNA and protein expression of cytochrome P450 3A in cultured rat primary hepatocyte.p><p>METHODRat heaptocyte were isolated by two-step in situ collagenase perfusion method; the hepatocytes were seeded in dishes coated with type I rat tail collagen, culture medium was added and changed daily after gelation, the effect of 18beta-glycyrrhizic acid and 18alpha-glycyrrhizic acid on the expression of CYP3A was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and western blot respectively.p><p>RESULTTreatment with 18beta-glycyrrhizic acid of primary rat hepatocytes resulted in marked up-regulation of CYP3A1 expression at both mRNA and protein levels in a concentration dependent manner, while exposure to 18alpha-glycyrrhizic acid of primary rat hepatocytes resulted in marked down-regulation of CYP3A1 expression at both mRNA and protein levels in a concentration dependent manmer.p><p>CONCLUSION18beta-glycyrrhizic acid and 18alpha-glycyrrhizic acid up-regulate and down-regulate CYP3A1 expression at the transcriptive levels.p>
Animals ; Clinical Laboratory Techniques ; Cytochrome P-450 CYP3A ; genetics ; metabolism ; Glycyrrhizic Acid ; pharmacology ; Hepatocytes ; drug effects ; metabolism ; Protein Biosynthesis ; drug effects ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Transcription, Genetic ; drug effects ; genetics