CYP1A1 mRNA Expression Research Articles

Quercetin Reduces the Development of 2,3,7,8-Tetrachlorodibenzo-p-dioxin-Induced Cleft Palate in Mice by Suppressing CYP1A1 via the Aryl Hydrocarbon Receptor.

Quercetin is a flavonoid with a wide range of pharmacological activities, including anticancer, antioxidant, and anti-inflammatory effects. Since it is a nutrient that can be consumed with a regular diet, quercetin has recently garnered interest. Quercetin acts as a phytochemical ligand for the aryl hydrocarbon receptor (AhR). Cleft lip and palate are among the most frequently diagnosed congenital diseases, and exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) during pregnancy induces cleft palate via AhR. In this study, we investigated the preventive effect of quercetin intake on the TCDD-induced cleft palate and its mechanism of action. The in vivo results suggest that quercetin intake by pregnant mice can prevent cleft palate in fetal mice. In vitro, the addition of TCDD induced a reduction in cell migration and the proliferation of mouse embryonic palatal mesenchymal cells, which was mitigated by the addition of quercetin. The addition of quercetin did not alter the mRNA expression levels of the AhR repressor but significantly suppressed mRNA expression of CYP1A1. In addition, the binding of AhR to a xenobiotic responsive element was inhibited by quercetin, based on a chemically activated luciferase expression assay. In conclusion, our results suggest that quercetin reduces the development of TCDD-induced cleft palate by inhibiting CYP1A1 through AhR.

In vitro CYP450 enzyme down-regulation by GLP-1/glucagon co-agonist does not translate to observed drug-drug interactions in the clinic.

NN1177 is a glucagon/glucagon-like peptide 1 receptor co-agonist investigated for chronic weight management and treatment of non-alcoholic steatohepatitis. Here, we show concentration-dependent down-regulation of cytochrome P450 enzymes using freshly isolated human hepatocytes treated with this linear 29-amino acid peptide. Notably, reductions in CYP3A4 mRNA expression (57.2-71.7%) and activity (18.5-51.5%) were observed with a clinically-relevant concentration of 100 nM NN1177. CYP1A2 and CYP2B6 were also affected, but to a lesser extent. Physiological-based pharmacokinetic modelling simulated effects on CYP3A4 and CYP1A2 probe substrates (midazolam and caffeine, respectively) and revealed potential safety concerns related to drug-drug interactions (DDIs). To investigate the clinical relevance of observed in vitro CYP down-regulation, a phase 1 clinical cocktail study was initiated to assess the DDI potential. The study enrolled 45 study participants (BMI 23.0-29.9 kg/m2) to receive a Cooperstown 5+1 cocktail (midazolam, caffeine, omeprazole, dextromethorphan, and S-warfarin/vitamin K) alone and following steady state NN1177 exposure. The analysis of pharmacokinetic profiles for the cocktail drugs showed no significant effect from the co-administration of NN1177 on AUC0-inf for midazolam or S-warfarin. Omeprazole, caffeine, and dextromethorphan generally displayed decreases in AUC0-inf and Cmax following NN1177 co-administration. Thus, the in vitro observations were not reflected in the clinic. These findings highlight remaining challenges associated with standard in vitro systems used to predict DDIs for peptide-based drugs as well as the complexity of DDI trial design for these modalities. Overall, there is an urgent need for better pre-clinical models to assess potential drug-drug interaction risks associated with therapeutic peptides during drug development. Significance Statement This study highlights significant challenges associated with assessing drug-drug interaction risks for therapeutic peptides using in vitro systems, since potential concerns identified by standard assays did not translate to the clinical setting. Further research is required to guide investigators involved in peptide-based drug development towards better non-clinical models in order to more accurately evaluate potential drug-drug interactions.

Cytochrome P450 inhibition activities of non-standardized botanical products

Ethnopharmacological relevanceR-tab, H-tab and E-cap botanical products are used for the treatment of various ailments. R-tab is traditionally prescribed for improving urination, H-tab is for relieving piles, hemorrhoids, fissures, and rectal inflammation and E-cap is for regulating menstruation.Aims of the study: To extract the botanical products and determine their potential interaction with the cytochrome P450 (CYP1A2, CYP2D6 and CYP3A4) enzymes. Materials and methodsR-tab, H-tab and E-cap botanical products were first extracted using solvents and analyzed using HPLC and LC-MS/MS. The effects of methanol extracts on the cytochrome induction and inhibition activities were determined using a series of in vitro assays, including multiplex RT-qPCR, CYP activity assays (P450-Glo™) and LC-MS/MS-based assays. For the CYP induction assay, omeprazole, rifampicin and dexamethasone were used as CYP1A2, CYP2D6 and CYP3A4 inducers, respectively. Ketoconazole and acetaminophen were used as positive and negative controls for the CYP3A4 inhibition assay, whereas furafylline and ketoconazole were used as positive and negative controls for the CYP1A2 inhibition assay. ResultsAll three botanical products did not show any significant induction in CYP1A2, CYP2D6 and CYP3A4 mRNA expression. By contrast, R-tab inhibited the mRNA expression of CYP1A2 significantly from the lowest concentration of 0.01 μg/mL, while, H-tab inhibited the mRNA expression of CYP1A2 and CYP3A4 from 0.1 μg/mL. Based on the P450 Glo assays, E-cap extract inhibited the metabolic activity of CYP1A2 with an IC50 value of 37.24 μg/mL. On the other hand, R-tab, H-tab and E-cap showed inhibitory effects on the CYP3A4 enzymatic activity with IC50 values of 17.42, 18.20 and 20.60 μg/mL, respectively. However, using the LC-MS/MS-based methods, the concentration-dependent effects of R-tab and H-tab extracts on the metabolism of testosterone appeared to be more prominent, with IC50 values of 51.90 and 56.90 μg/mL as compared with the rest of the results, which were all above 100 μg/mL ConclusionThe CYP3A4 mRNA and enzymatic activity were moderately inhibited by R-tab and H-tab. Methanol extract of botanical products in solid dosage forms can be evaluated for their herb-drug interaction risks using in vitro assays and may provide the minimum data required for safety labeling.

Liquiritin ameliorates metabolic and endocrine alterations in a mouse model of polycystic ovary syndrome

Abstract Objective: Altered bile acid transformation induces low-grade chronic inflammation and may play an important role in the pathophysiology of polycystic ovary syndrome (PCOS). Liquiritincan regulate bile acid metabolism and anti-inflammatory properties; however, limited information is available regarding its therapeutic potential in PCOS. Methods: Female C57BL/6 mice were randomly assigned into four groups (n = 6 mice/group): the control, letrozole or dehydroepiandrosterone-induced PCOS groups, PCOS + 20 mg/kg liquiritin group, and control + liquiritin groups. After 21 days of treatment, the mice were euthanized, and the associated metabolism indications were investigated. Ovarian histological examinations were performed, and serum hormone concentration was measured. The expression of key genes involved in steroid hormone synthesis, ovarian follicle development, and ovulation was assessed. Results: Liquiritin reduced fasting blood glucose levels and increased insulin sensitivity compared to the PCOS group. Liquiritin also significantly decreased serum levels of total testosterone (P < 0.001) and dehydroepiandrosterone sulfate (P < 0.05) in the PCOS group. Histomorphological inspection of ovaries from the liquiritin group revealed fewer cystic dilated follicles than in the PCOS group. Moreover, liquiritinsignificantly (P < 0.01) decreased Cyp17a1, Cyp19a1, Fshr, Hsd3b2, Runx2, and Ccn2 mRNA expression compared to letrozole-induced PCOS. Conclusion: Liquiritin may be safe and helpful in ameliorating PCOS-associated hyperandrogenemia and hyperglycemia. However, clinical trials investigating different liquiritin dosages are needed to confirm these findings.

Curcumin and quercetin modify warfarin-induced regulation of porcine CYP1A2 and CYP3A expression and activity in vitro

The anticoagulant drug warfarin is used treat atrial fibrillation. Several cases of drug-drug and drug-food interactions has been reported for warfarin. The aim of the current study, were to investigate the interaction between simultaneous administration of warfarin with the two ubiquitous flavonoids quercetin and curcumin. Using porcine primary hepatocytes we demonstrated that warfarin treatment increased the mRNA and protein expression of CYP3A(29), while no changes in CYP1A2 were observed. Co-treatment with quercetin and/or curcumin decreased the warfarin induced CYP3A protein expression. Moreover, when quercetin and curcumin were co-administrated to warfarin-exposed hepatocytes the protein expression of CYP1A2 were decreased. In hepatic microsomes, curcumin inhibited the activity of both CYP1A2 and CYP3A, while warfarin had no effect. Both quercetin and curcumin decreased the CYP1A2 and CYP3A activity when co-administrated with warfarin. The results clearly demonstrated that quercetin and curcumin can cause food-drug interactions with warfarin, and that the cocktail effect of exposure to more compounds than one can further enhance these interactions.

Alterations in circadian rhythms aggravate Acetaminophen-induced liver injury in mice by influencing Acetaminophen metabolization and increasing intestinal permeability

ABSTRACT Acetaminophen (APAP) is the most common antipyretic and analgesic drug causing drug-induced liver injury (DILI). Alterations in circadian rhythms can adversely affect liver health, especially metabolic and detoxification functions. However, the effect of circadian rhythm alterations induced by environmental factors on APAP-induced liver injury and the underlying mechanisms are not well known. In this study, a mouse model of circadian rhythm alterations was established by light/dark cycle shift and then treated with excessive APAP. The liver injury indexes, APAP-related metabolic enzymes, and intestinal permeability in mice were evaluated by biochemical analysis, quantitative real-time PCR, enzyme-linked immunosorbent assays, and histopathology. Results showed that circadian rhythm alterations resulted in increased reactive oxygen species (ROS) and malondialdehyde (MDA) and decreased liver superoxide dismutase (SOD), glutathione, and CYP1A2 and CYP3A11 mRNA expression, and increased serum diamine oxidase, lipopolysaccharide, and D-lactate in the mice. Compared with control mice, APAP induced higher serum alanine aminotransferase and aspartate aminotransferase, liver interleukin-1β and tumor necrosis factor-α mRNA, ROS and MDA, lower SOD, glutathione, and UDP-glucuronosyltransferases /sulfotransferases mRNA and more severe liver necrosis and intestinal damage in mice with alterations in circadian rhythms. In conclusion, circadian rhythm alterations by light/dark cycle shift resulted in increased oxidative stress and intestinal permeability in the mice and exacerbated APAP-induced liver injury by influencing APAP metabolization and increasing intestinal permeability.

STING is an intrinsic checkpoint inhibitor that restrains the TH17 cell pathogenic program.

SUMMARYExternal and intrinsic factors regulate the transcriptional profile of T helper 17 (TH17) cells, thereby affecting their pathogenic potential and revealing their context-dependent plasticity. The stimulator of interferon genes (STING), a component of the intracellular DNA-sensing pathway, triggers immune responses but remains largely unexplored in T cells. Here, we describe an intrinsic role of STING in limiting the TH17 cell pathogenic program. We demonstrate that non-pathogenic TH17 cells express higher levels of STING than those activated under pathogenic conditions. Activation of STING induces interleukin-10 (IL-10) production in TH17 cells, decreasing IL-17A and IL-23R expression in a type I interferon (IFN)-independent manner. Mechanistically, STING-induced IL-10 production partially requires aryl hydrocarbon receptor (AhR) signaling, while the decrease of IL-17A expression occurs due to a reduction of Rorγt transcriptional activity. Our findings reveal a regulatory function of STING in the TH17 cell activation program, proposing it as a valuable target to limit TH17-cell-mediated inflammation.

Influence of High‐Polyphenol Sorghum on Benzo[a]pyrene‐Metabolizing Enzymes

Colorectal cancer ranks among the top five most common types of cancer in the United States. Lifestyle, diet, and environmental factors are correlated to the increased risk of colon cancer. One of those environmental factors is the polycyclic aromatic hydrocarbon (PAHs) Benzo[a]pyrene (BaP), formed by the incomplete combustion of organic material. Human exposure to BaP occurs through air pollution, tobacco smoke, engine exhaust, and the consumption of BaP‐exposed foods. BaP crosses cellular membranes, and it becomes metabolically activated by Phase I enzymes, primarily cytochrome P450s (CYPs) and Epoxide Hydrolase, and subsequently has the potential to bind to DNA. The binding of BaP to DNA may result in adverse genotypic alterations, which may result in tumorigenesis. On the other hand, cellular Phase II enzymes play a role in the inactivation of BaP and its metabolites, which can facilitate the excretion of these molecules. Polyphenols, a group of plant micronutrients known for their antioxidant and anti‐inflammatory properties, have been suggested to contribute to the prevention of colon cancer. Extracts from two types of Sorghum bicolor were provided by the USDA ARS Kansas. Preliminary studies in our lab suggest that polyphenol‐rich varietals of the grain Sorghumaffect pathways in carcinogenesis. Specifically, mRNA expression of UDP‐glucuronosyl transferase (UGT) 1A1, a Phase II PAH‐bioinactivating enzyme, is significantly increased in high‐polyphenol sorghum treated HCT116 cells. Therefore, we propose to investigate the impact of high‐polyphenol sorghum on BaP‐exposed colon cancer cells, assessing the expression and activity of Phase I and Phase II enzymes in particular. Our hypothesis is that compared to controls, high polyphenol sorghum extracts will decrease expression and activity of Phase I, and increase expression and activity of Phase II enzymes in BaP‐treated colon cancer cells.Human colorectal cancer HCT116 cells were incubated with either 0, 1, 2, or 5 µM BaP alone, Sorghum extracts alone, or in the presence of high‐polyphenol sorghum and Benzo[a] pyrene for up to 48 hours. RNA was extracted and reverse‐transcribed to cDNA. mRNA expression of carcinogen activating and inactivating enzymes was measured using qPCR. BaP alone increased mRNA expression of CYP1A1 as expected. Our high polyphenol Sorghum extract dramatically decreased mRNA expression of the Phase I enzyme CYP1B1, but also moderately increased mRNA expression of Epoxide Hydrolase. We plan to complete assessment of the expression and catalytic activity of Phase I and Phase II enzymes in the presence of high‐polyphenol sorghum and Benzo[a] pyrene. Our initial results suggest that high‐polyphenol Sorghum may be able to shift the balance of Phase I and Phase II enzymes in BaP‐exposed cells, and thus provide an avenue of a dietary approach to support chemoprevention.

Expression of Cytochrome P450 Enzymes in Pediatric Non-Rhabdomyosarcoma Soft Tissue Sarcomas: Possible Role in Carcinogenesis and Treatment Response.

The 5-year relative survival rate estimate of treated patients with non-rhabdomyosarcoma soft tissue sarcomas (NRSTS) is ∼50% since they generally present with tumor progression, relapse, metastasis, and/or chemoresistance. The expression of cytochrome P450 (CYP) enzymes in malignancies can affect the pharmacology of drugs commonly used in chemotherapy or confer susceptibility to development of chemical carcinogenesis; in addition, their specific tumor expression can be used as a therapeutic target. Using qPCR and Western blot assays, the expression of CYP1B1, CYP2E1, CYP3A4, and CYP3A5 were analyzed in a cohort of tumor tissue paired with non-malignant adjacent tissue of patients with NRSTS. The mRNA and protein expression of CYP1B1, CYP2E1, and CYP3A4 were significantly increased in tumor tissue. We propose that the expression of these isoforms is related to carcinogenesis and chemoresistance frequently observed in these neoplasms.

Bird’s eye view analysis of in situ cholesterol metabolic pathways in breast cancer patients and its clinicopathological significance in their subtypes

Obesity has been known to increase the risks of breast cancer (BC) development and also to be associated with adverse clinical outcome of the patients. Abnormalities of cholesterol metabolism are not only related to obesity but also to biological or clinical behavior of BC patients. However, which metabolites or pathways of cholesterol metabolism could represent the characteristics of BC patients have remained virtually unknown. Therefore, in this study, we attempted to perform bird’s eye view or comprehensive analysis of in situ or intra-tumoral cholesterol metabolic pathways using the multimodal approaches in order to elucidate the possible significance of cholesterol metabolites and its metabolic enzymes including CYP27A1, CYP7A1, and CYP46A1. GC-MS study using BC specimens was first performed in 60 BCE patients to evaluate cholesterol metabolism from cholesterol through oxysterols in both BC and normal tissues. Results of those analyses above lead to evaluating immunoreactivity and mRNA expression of CYP27A1, CYP7A1 and CYP46A1 in 213 and 153 BCE cases, respectively. Results of comprehensive GC-MS analysis did reveal that three oxysterols, 27-HC, 7α-HC and 24-HC were all related to malignant phenotypes in BC. 27-HC abundance was significantly associated with higher tumor stage (P = 0.0475) of BC patients. Luminal B type BC patients harboring high CYP27A1, the enzyme responsible for production of 27-HC were significantly associated with worse disease-free survival than those with low CYP27A1 (P = 0.0463). 7α-HC tended to be more abundant in HER2 positive and TNBC subtypes and higher levels of 7α-HC were also significantly associated with higher Ki-67 labeling index (P = 0.0022) and histological grade (P = 0.0286). CYP7A1, the enzyme involved in production of 7α-HC, was significantly more abundant in TNBC than other subtypes (vs Luminal A; P = 0.0321, vs Luminal B; P = 0.0048, vs HER2; P = 0.0103). The levels of 24-HC in BC were lower than normal breast tissues regardless of its subtypes. CYP46A1, the enzyme involved in the production of 24-HC, was detected only in 33 (15.5%) out of 213 BCE cases examined in this study. Results of our bird’s eye view analysis of in situ or intra-tumoral cholesterol metabolism in BC patients did firstly reveal BC subtype dependent involvement of its different pathways. Results also indicated the therapeutic possibility of subtype dependent modification of cholesterol metabolizing pathways in BC patients.

Apelin and Apelin Receptor in Follicular Granulosa Cells of Buffalo Ovaries: Expression and Regulation of Steroidogenesis.

Apelin (APLN), as a ligand for APJ (an orphan G-protein-coupled receptor), is an adipokine with pleiotropic effects in many physiological processes of the body. It has an important role in the control of reproduction particularly in females (mainly in control of ovarian function). This study was carried out to investigate the mRNA and protein amounts of APLN/APJ in granulose cells (GCs) of ovarian follicles with small (SF), medium (MF), and large (LF) sizes of buffalo (Bubalus bubalis) and the effect of IGF1 and follicle-stimulating hormone (FSH) on the expression levels of APLN/APJ. In addition, we evaluated the effect of various doses of APLN (isoforms -13 and -17) singly or in combination with IGF1 and FSH on estradiol (E2) and progesterone (P4) secretion in GCs. The mRNA and protein abundance of APLN was the highest in GCs of LF while the APJ expression enhanced with follicle enlargement in GCs (p-value <0.01). IGF1 and FSH elevated the mRNA and protein amounts of APLN and FSH, and IGF1 increased the expression of APJ in buffalo GCs (p-value <0.01). Both isoforms of APLN (-13/-17) singly or in the presence of IGF1 or FSH increased the secretion of E2 and P4 with or without preincubation of cells with APJ antagonist (ML221 10 µM), although we had some variation in the effects. Concurrently, APLN-13/-17 significantly increased the mRNA and protein expression of CYP19A1 and StAR (p-value <0.01). ML221 substantially diminished the secretion of E2 and P4 and also the expression of CY19A1 and StAR in buffalo GCs (p-value <0.01). We also revealed that APLN-13/-17 (10-9 M), singly or in response to IGF1 and FSH, increased the production of E2 and P4 in different times of stimulation. In conclusion, APLN may play a crucial role in steroidogenesis and follicular development in ovarian GCs of buffalo.

Gene Expression Profiling of 1α,25(OH)2 D3 Treatment in 2D/3D Human Hepatocyte Models Reveals CYP3A4 Induction but Minor Changes in Other Xenobiotic-Metabolizing Genes.

CYP3A4 is the most important drug-metabolizing enzyme regulated via the vitamin D receptor(VDR) in the intestine. However, less is known about VDR in the regulation of CYP3A4 and other drug-metabolizing enzymes in the liver. This study investigates whether 1α,25-dihydroxyvitamin D3 (1α,25(OH)2 D3 ) regulates major cytochrome P450 enzymes, selected phase I and II enzymes, and transporters involved in xenobiotic and steroidal endobiotic metabolism in 2D and 3D cultures of human hepatocytes. The authors found that 1α,25(OH)2 D3 increases hepatic CYP3A4 expression and midazolam 1'-hydroxylation activity in 2D hepatocytes. The results are confirmed in 3D spheroids, where 1α,25(OH)2 D3 has comparable effect on CYP3A4 mRNA expression as 1α-hydroxyvitamin D3 , an active vitamin D metabolite. Other regulated genes such as CYP1A2, AKR1C4, SLC10A1, and SLCO4A1 display only mild changes in mRNA levels after 1α,25(OH)2 D3 treatment in 2D hepatocytes. Expression of other cytochrome P450, phase I and phase II enzyme, or transporter genes are not significantly influenced by 1α,25(OH)2 D3 . Additionally, the effect of VDR activation on CYP3A4 mRNA expression is abolished by natural dietary compound sulforaphane, a common suppressor of pregnane X receptor(PXR) and constitutive androstane receptor(CAR). This study proposes that VDR or vitamin D supplementation is unlikely to significantly influence liver detoxification enzymes apart from CYP3A4.

CYP1A2 mRNA Expression Rather than Genetic Variants Indicate Hepatic CYP1A2 Activity.

CYP1A2, one of the most abundant hepatic cytochrome P450 enzymes, is involved in metabolism of several drugs and carcinogenic compounds. Data on the significance of CYP1A2 genetic polymorphisms in enzyme activity are highly inconsistent; therefore, the impact of CYP1A2 genetic variants (−3860G>A, −2467delT, −739T>G, −163C>A, 2159G>A) on mRNA expression and phenacetin O-dealkylation selective for CYP1A2 was investigated in human liver tissues and in psychiatric patients belonging to Caucasian populations. CYP1A2*1F, considered to be associated with high CYP1A2 inducibility, is generally identified by the presence of −163C>A polymorphism; however, we demonstrated that −163C>A existed in several haplotypes (CYP1A2*1F, CYP1A2*1L, CYP1A2*1M, CYP1A2*1V, CYP1A2*1W), and consequently, CYP1A2*1F was a much rarer allelic variant (0.4%) than reported in Caucasian populations. Of note, −163C>A polymorphism was found to result in an increase of neither mRNA nor the activity of CYP1A2. Moreover, hepatic CYP1A2 activity was associated with hepatic or leukocyte mRNA expression rather than genetic polymorphisms of CYP1A2. Consideration of non-genetic phenoconverting factors (co-medication with CYP1A2-specific inhibitors/inducers, tobacco smoking and non-specific factors, including amoxicillin+clavulanic acid therapy or chronic alcohol consumption) did not much improve genotype–phenotype estimation. In conclusion, CYP1A2-genotyping is inappropriate for the prediction of CYP1A2 function; however, CYP1A2 mRNA expression in leukocytes can inform about patients’ CYP1A2-metabolizing capacity.

Sorafenib is an antagonist of the aryl hydrocarbon receptor

Sorafenib is an orally administered inhibitor of several tyrosine protein kinases. Treatment with sorafenib induces autophagy, which may suppress the growth of hepatocellular carcinoma (HCC) and other cancers. Aryl hydrocarbon receptor (AhR) is activated by xenbiotics and is involved in detoxification, but also plays other physiological roles. The following results were obtained. ITE and β-NF are endogenous and synthetic AhR ligands, respectively. One μM sorafenib can strongly suppress baseline as well as 0.5 μM ITE- and 1 μM β-NF-induced transcriptional activity of the aryl hydrocarbon response element (AHRE) in both human and mouse cells. Cytochrome p450 (CYP) 1A1 is mainly transcribed by activated AhR. Sorafenib (2−15 μM) strongly and dose-dependently suppressed baseline as well as 2 μM ITE- and 10 μM β-NF-induced CYP1A1 mRNA and protein expression. Ligand-activated AhR translocates from the cytoplasm to the nucleus. While sorafenib was found to suppress AhR activity, the drug alone was able to induce AhR translocation into the nucleus. Sorafenib’s antagonistic action on AhR was comparable to that of the known AhR antagonist CH-223191 in human liver and ovarian cell lines. In summary, we demonstrate that sorafenib is a potent AhR antagonist and likely endocrine disruptor of the AhR. Moreover, sorafenib offers potential benefit for diseases treatable through AhR suppression strategies. Further investigation is warranted into sorafenib’s AhR antagonistic behavior.

Gene expression of glutathione S-transferase alpha, glutathione S-transferase rho, glutathione peroxidase, uncoupling protein 2, cytochrome P450 1A, heat shock protein 70 in liver of Oreochromis niloticus upon exposure of microcystin-LR, microcystin-RR and toxic cyanobacteria crude

ObjectiveMicrocystins (MCs) are considered to be one of the most hazard groups of cyanobacteria and are known as a group of cyclic polypeptide hepatotoxins of varying potency. The study determined the variation of gene expression of GSTA, GSTR, GPX, UCP2, CYP1A1 and HSP70 genes in liver of tilapia fish (Oreochromis niloticus) under the effect of MC-LR or MC-RR toxins. MethodsTilapia fish adult male fishes (n = 20; 120–178 g) obtained from fish farm. Fishes injected i.p. with a single dose of 50 μg/kg MC-LR, and 20 fishes injected with 500 μg/kg MC-RR. Whereas the control group was only received the vehicle solution. Total mRNA was isolated from 50 mg of hepatic sections. The relative levels of GSTA, GSTR, GPX, UCP2, HSP70/β actin mRNA were determined by RT-real time PCR. ResultsSignificantly increased in GSTA mRNA expression was observed in the liver of fish after exposure to all exposure groups. Variable alterations in the levels of GPx, GSTR, UCP2 and CYP1A mRNA expression were observed post exposure groups. Whereas, the relative level of HSP70 mRNA was increased at 12 h after MCs exposure and was elevated gradually at 24 h until 7 days post treatment. ConclusionThe presence of inducible expression of GSTA, GSTR and GPx, and no significant changes of CYP1A mRNA expression in the liver of tilapia post MCs exposure, indicates that it should be the phase II detoxification enzyme in the liver of tilapia, but not the phase I detoxification enzyme. In addition to restrain over production of ROS, increasing of UCP2 and HSP70 expression may play an important role as a molecular chaperone against oxidative stress consequently, preventing hepatic cells from apoptosis.

Alteration of murine cytochrome P450 profiles in fatty liver disease by hesperidin and myricetin

Background: Hesperidin and myricetin are anti-inflammatory and anti-oxidant flavonoids that have beneficial effects in fatty liver disease (FLD), but information regarding their effects on cytochrome P450 (CYP450) enzymes in FLD is limited. Objectives: This study determined the impacts of hesperidin and myricetin on CYP450 profiles in FLD mice. Materials and Methods: Adult female mice were fed a high fat diet (HFD, 60 kcal % fat of total food) with daily intragastrically administered ethanol (0.5 g/kg/day) in combination with either fenofibrate (40 mg/kg/day), hesperidin (50 and 200 mg/kg/day), or myricetin (50 and 200 mg/kg/day) for 60 consecutive days. Liver histomorphology was examined by oil red O staining. Hepatic enzyme activity and mRNA expression of CYP1A2, CYP2E1, CYP3A11, CYP3A13, and CYP4A11 were assessed. Results: HFD-induced hepatocellular damage was prevented by low dose hesperidin and myricetin. Expression of Cyp1a2, Cyp2e1, and Cyp4a11 mRNAs as well as CYP2E1 and CYP3A activities was significantly induced by HFD plus ethanol (HE) while Cyp3a13 expression was slightly increased. Fenofibrate and low dose hesperidin prevented HE-induced Cyp1a2 and Cyp2e1 expression while HE-induced Cyp4a11 expression was prevented by all treatments. The expression of Cyp3a13 was extensively suppressed by high-dose hesperidin and myricetin, and CYP2E1 and CYP3A activities were significantly decreased by all treatments. Conclusion: Alteration of CYP450 profiles by high doses of hesperidin and myricetin could lead to drug interactions. Nevertheless, low dose hesperidin prevented dysregulation of CYP450 expression in FLD and is a promising candidate for FLD treatment.

Dietary Resveratrol Alleviates AFB1-Induced Ileum Damage in Ducks via the Nrf2 and NF-κB/NLRP3 Signaling Pathways and CYP1A1/2 Expressions

The aim of this study was to explore the mechanism underlying the protective effects of resveratrol against Aflatoxin B1-induced ileum injury in ducks. A corn–soybean meal-basal diet and two test diets (500 mg/kg resveratrol +0.2 mg Aflatoxin B1/kg, 0.2 mg AFB1/kg) were used in a 10-wk design trial (n = 15 ducks/group). These results showed that the toxicity of Aflatoxin B1 significantly reduced the antioxidant capacity of duck ileum and induced inflammation, oxidative stress, mitochondrial dysfunction and DNA damage in ducks. The expression of genes, including CYP1A2, CYP2A6, and CYP3A4, at the mRNA level was significantly upregulated (p &lt; 0.05) by AFB1. The level of Nrf2 was suppressed (p &lt; 0.05) and the mRNA and protein level of NF-κB was activated (p &lt; 0.05) in the AFB1 group. However, supplementation with 500 mg/kg dietary resveratrol in Aflatoxin B1-induced ducks significantly ameliorated these alterations and decreased the mRNA expression of CYP1A1 and CYP1A2 (p &lt; 0.05) and the production of AFB1-DNA adducts (p &lt; 0.05). The results proved that resveratrol alleviated ileum injury induced by AFB1, decreased the production of AFB1-DNA adducts by downregulating the expression of CYP1A1 and CYP1A2, and reduced DNA damage and oxidative stress via the Nrf2/ Keap1 and NF-κB/NLRP3 signaling pathways.

Effect of total flavonoids of Hippophae rhamnoides L. on the activity and mRNA expression of CYP450 in rats

Objectives: In this study, we aimed to study the effect of total flavonoids extracted from Hippophae rhamnoides L. (TFH) on the activity of five CYP450 enzymes in rats by high-performance liquid chromatography (HPLC). We also studied the effect of TFH on the expression of CYP450 enzyme mRNA in rat liver using quantitative real-time PCR (qRT-PCR). Materials and Methods: TFH was orally administered to male rats once daily at doses of 100, 200, and 400 mg/kg/day for 14 days. We established HPLC method to detect the concentration of the analytes in the rat plasma. We used DAS software to calculate the pharmacokinetic parameters. Then, qRT-PCR was used to study the effect of TFH on the expression of CYP450 mRNA. Results: The pharmacokinetic analysis of theophylline in the TFH group revealed a short half-life (t1/2), a decrease in the area under the curve (AUC), and an increase in the value of plasma clearance (CL) (P < 0.05, P < 0.01). In the case of midazolam, t1/2 was prolonged, Cmax was increased (P < 0.05). Results from the PCR analysis indicate that TFH downregulated the expression of CYP3A4 mRNA and upregulated the expression of CYP1A2 mRNA. Conclusion: In conclusion, TFH might induce the activity of CYP1A2 and inhibit the activity of CYP3A4. Herb–drug interaction should be carefully considered when TFH is being used in combination with drugs metabolized by CYP1A2 and CYP3A4.SUMMARYSeabuckthorn is a kind of drug and food homologous substance. Care should be taken when using drugs metabolized by CYP1A2 and CYP3A4 in combination.[INLINE:1]Abbreviations used: TFH: Total flavonoids of Hippophae rhamnoides L.; t1/2: Half-life; CYP450: Cytochrome P450; IS: Internal standard; AUC: Area under the curve; qRT-PCR: Quantitative Real-time PCR; CL: Plasma clearance; DDIs: Drug-drug interactions; Cmax: Maximum plasma concentration.

An In Vitro Study on the Effects of Aromatic Hydrocarbons 1- Naphthol and Dibenz[a,h]anthracene in HepG2 Cells

Liver cells (HepG2) were treated with well known toxic aromatic hydrocarbons 1-Naphthol and Dibenz[a,h]anthracene. Naphthol at 20-40µg/ml affected significant increase in the levels of reactive oxygen species in the cells.. Reactive nitrogen intermediates increased to an extent of 2.4 to 3.8 fold in cells treated with naphthol over a range of 20-80 µg/ml. Expression of antioxidant enzyme Glutathione peroxidase was found to increase two fold, while catalase expression was found to decrease by 21% on exposure to 1-Naphthol. Dibenz[a,h]anthracene did not affect discernible changes in the expression of these enzymes Induction of CYP1A1 mRNA by Naphthol at 60µg/ml was around 45 fold higher than untreated cells. Dibenz anthracene at a dose of 5µg/ml was found to be a potent inducer of CYP1A1 as it elevated expression of CYP1A1 mRNA to an extent of 350 fold. . Naphthol exhibited lipogenic potential in HepG2 cells. Concommitant increase in Sterol regulatory element binding protein 1c was observed.

التأثيرات الناتجة عن استخدام عقاري الديكلوفيناك والسيبروفلوكساسين على الكبد والكلى لدى الجرذان

The toxic effect of diclofenac (DCF) sodium and Ciprofloxacin (CIP) on gene expression of cytochrome P450 oxidase (CYPs) and the histology of liver and kidney of male albino rat has been evaluated in this study. DCF and CIP were chosen since they are inhibitors for specific CYP enzymes. Thirty-five adult male albino rats were divided into 7 groups of 5 animals each (A, B, C, D, E, F and G) and were treated orally with drugs for 21 consecutive days. Group A served as the control while B and C were treated with 5.3, 10.6 mg/kg body weight (bw) DCF sodium and groups D and E were treated with 40 and 80 mg/kg bw CIP, respectively. Groups F and G were treated with a mixture of the low and the high doses of both drugs, respectively. Both drugs significantly downregulated the mRNA expression of CYP1a2, CYP3a4 and CYP2c9. They caused hepatorenal histological changes. In the liver, massive fibrosis, necrosis, inflammatory cell infiltration with hemorrhages and hydrophilic degeneration have been observed. A massive tissue injury with glomerular and tubular damages due to sever necrosis, degeneration of concomitant inflammatory cells and blood vessels congestion have been shown in renal tissues. Although DCF and CIP are still used as therapeutic drugs, their use should be limited as their chronic administration induces a toxic effect on human health.