scholarly journals The reference liver—CYP450 and UGT enzymes in healthy donor and metastatic livers: the impact of genotype

2021 ◽  
Author(s):  
Mateusz Kurzawski ◽  
Sylwia Szeląg-Pieniek ◽  
Joanna Łapczuk-Romańska ◽  
Maciej Wrzesiński ◽  
Stefan Oswald ◽  
...  
Keyword(s):  
Protein Content ◽  
Mrna Level ◽  
Inverse Correlation ◽  
Control Group ◽  
Drug Metabolizing Enzymes ◽  
Protein Abundance ◽  
Metabolizing Enzymes ◽  
Functional Variants ◽  
Common Genetic Variants ◽  
The Impact

Abstract Background Hepatic enzymes involved in drug metabolism vary markedly in expression, abundance and activity, which affects individual susceptibility to drugs and toxicants. The present study aimed to compare mRNA expression and protein abundance of the most pharmacologically relevant drug-metabolizing enzymes in two main sources of the control liver samples that are used as the reference, i.e. organ donor livers and non-tumorous tissue from metastatic livers. An association analysis of the most common genetic variants with mRNA and protein levels was also performed. Methods The CYP450 and UGT enzymes (CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, CYP3A5, UGT1A1, UGT1A3, UGT2B7 and UGT2B15) were analyzed for mRNA (qPCR) and protein abundance (LC–MS/MS) in healthy donors (n = 11) and metastatic (n = 13) livers. Genotyping was performed by means of TaqMan assays and pyrosequencing. Results Significantly higher protein abundance in the metastatic livers was observed in case of CYP2C9, CYP2D6, and UGT2B7, and for UGT1A3 the difference was only significant at mRNA level. For all the enzymes except CYP2E1 some significant correlation between mRNA and protein content was observed, and for UGT1A1 an inverse correlation with age was noted. CYP2C19, CYP3A5 and CYP2D6 were significantly affected by genotype. Conclusion The selection of a control group for the study on drug-metabolizing enzymes (e.g. in pathological states) may possibly affect its conclusions on differences in mRNA and protein content. Genotyping for common functional variants of CYP450 enzymes should be performed in all studies on drug-metabolizing enzymes.

scholarly journals Hepatic drug-metabolizing enzymes and drug transporters in Wilson’s disease patients with liver failure

2021 ◽  
Author(s):  
Sylwia Szeląg-Pieniek ◽  
Stefan Oswald ◽  
Mariola Post ◽  
Joanna Łapczuk-Romańska ◽  
Marek Droździk ◽  
...  
Keyword(s):  
Gene Expression ◽  
Liver Failure ◽  
Wilson’S Disease ◽  
Drug Transporters ◽  
Wilson's Disease ◽  
Control Group ◽  
Drug Metabolizing Enzymes ◽  
Protein Abundance ◽  
Copper Transporter ◽  
Metabolizing Enzymes

Abstract Background Wilson’s disease is a genetic disorder inherited in a recessive manner, caused by mutations in the copper-transporter ATP7B. Although it is a well-known disease, currently available treatments are far from satisfactory and their efficacy varies in individual patients. Due to the lack of information about drug-metabolizing enzymes and drug transporters profile in Wilson’s disease livers, we aimed to evaluate the mRNA expression and protein abundance of selected enzymes and drug transporters in this liver disorder. Methods We analyzed gene expression (qPCR) and protein abundance (LC–MS/MS) of 14 drug-metabolizing enzymes and 16 drug transporters in hepatic tissue from Wilson’s disease patients with liver failure (n = 7, Child–Pugh class B and C) and metastatic control livers (n = 20). Results In presented work, we demonstrated a downregulation of majority of CYP450 and UGT enzymes. Gene expression of analyzed enzymes ranged between 18 and 65% compared to control group and significantly lower protein content of CYP1A1, CYP1A2, CYP2C8, CYP2C9, CYP3A4 and CYP3A5 enzymes was observed in Wilson’s disease. Moreover, a general decrease in hepatocellular uptake carriers from SLC superfamily (significant at protein level for NTCP and OATP2B1) was observed. As for ABC transporters, the protein abundance of BSEP and MRP2 was significantly lower, while levels of P-gp and MRP4 transporters were significantly higher in Wilson’s disease. Conclusions Altered hepatic expression of drug‐metabolizing enzymes and drug transporters in Wilson’s disease patients with liver failure may result in changes of drug pharmacokinetics in that group of patients.

scholarly journals A Pilot Study towards the Impact of Type 2 Diabetes on the Expression and Activities of Drug Metabolizing Enzymes and Transporters in Human Duodenum

2019 ◽  
Vol 20 (13) ◽  
pp. 3257 ◽  
Author(s):  
Sophie Gravel ◽  
Benoit Panzini ◽  
Francois Belanger ◽  
Jacques Turgeon ◽  
Veronique Michaud
Keyword(s):  
Type 2 Diabetes ◽  
Mrna Expression ◽  
Drug Metabolizing Enzymes ◽  
Mrna Levels ◽  
Metabolizing Enzymes ◽  
Expression Levels ◽  
The Impact ◽  
Duodenal Biopsies ◽  
Mrna Expression Levels

To characterize effects of type 2 diabetes (T2D) on mRNA expression levels for 10 Cytochromes P450 (CYP450s), two carboxylesterases, and three drug transporters (ABCB1, ABCG2, SLCO2B1) in human duodenal biopsies. To compare drug metabolizing enzyme activities of four CYP450 isoenzymes in duodenal biopsies from patients with or without T2D. mRNA levels were quantified (RT-qPCR) in human duodenal biopsies obtained from patients with (n = 20) or without (n = 16) T2D undergoing a scheduled gastro-intestinal endoscopy. CYP450 activities were determined following incubation of biopsy homogenates with probe substrates for CYP2B6 (bupropion), CYP2C9 (tolbutamide), CYP2J2 (ebastine), and CYP3A4/5 (midazolam). Covariables related to inflammation, T2D, demographic, and genetics were investigated. T2D had no major effects on mRNA levels of all enzymes and transporters assessed. Formation rates of metabolites (pmoles mg protein−1 min−1) determined by LC-MS/MS for CYP2C9 (0.48 ± 0.26 vs. 0.41 ± 0.12), CYP2J2 (2.16 ± 1.70 vs. 1.69 ± 0.93), and CYP3A (5.25 ± 3.72 vs. 5.02 ± 4.76) were not different between biopsies obtained from individuals with or without T2D (p > 0.05). No CYP2B6 specific activity was measured. TNF-α levels were higher in T2D patients but did not correlate with any changes in mRNA expression levels for drug metabolizing enzymes or transporters in the duodenum. T2D did not modulate expression or activity of tested drug metabolizing enzymes and transporters in the human duodenum. Previously reported changes in drug oral clearances in patients with T2D could be due to a tissue-specific disease modulation occurring in the liver and/or in other parts of the intestines.

scholarly journals Metabolic Acidosis Alters Expression of Slc22 Transporters in Mouse Kidney

2020 ◽  
Vol 45 (2) ◽  
pp. 263-274
Author(s):  
Janine Gottier Nwafor ◽  
Marta Nowik ◽  
Naohiko Anzai ◽  
Hitoshi Endou ◽  
Carsten A. Wagner
Keyword(s):  
Metabolic Acidosis ◽  
Mrna Level ◽  
The Body ◽  
Large Set ◽  
Protein Abundance ◽  
Waste Products ◽  
Anion Transporters ◽  
Solute Carrier ◽  
The Impact ◽  
Oxonic Acid

Introduction: The kidneys play a central role in eliminating metabolic waste products and drugs through transporter-mediated excretion along the proximal tubule. This task is mostly achieved through a variety of transporters from the solute carrier family 22 (SLC22) family of organic cation and anion transporters. Metabolic acidosis modulates metabolic and renal functions and also affects the clearance of metabolites and drugs from the body. We had previously shown that induction of metabolic acidosis in mice alters a large set of transcripts, among them also many transporters including transporters from the Slc22 family. Objective: Here we further investigated the impact of acidosis on Slc22 family members. Methods: Metabolic acidosis was induced for 2 or 7 days with NH4Cl, some animals also received the uricase inhibitor oxonic acid for comparison. Expression of transporters was studied by qPCR and immunoblotting. Results: NH4Cl induced no significant changes in plasma or urine uric acid levels but caused downregulation of Slc22a1 (Oct1), Slc22a6 (Oat1), Slc22a19 (Oat5), and ­Slc22a12 (Urat1) at mRNA level. In contrast, Slc22a4 mRNA (Octn1) was upregulated. On protein level, NH4Cl increased Octn1 (after 7 days) and Urat1 (after 2 days) abundance and decreased Oat1 (after 2 days) and Urat1 (after 7 days). Oxonic acid had no impact on protein abundance of any of the transporters tested. Conclusion: In summary, metabolic acidosis alters expression of several transporters involved in renal excretion of metabolic waste products and drugs. This may have implications for drug kinetics and clearance of waste metabolites.

The impact of genetic polymorphisms of drug metabolizing enzymes on the pharmacodynamics of clopidogrel under steady state conditions

2015 ◽  
Vol 30 (4) ◽  
pp. 295-304 ◽  
Author(s):  
Nontaya Nakkam ◽  
Somsak Tiamkao ◽  
Sirimas Kanjanawart ◽  
Siriporn Tiamkao ◽  
Suda Vannaprasaht ◽  
...  
Keyword(s):  
Steady State ◽  
Genetic Polymorphisms ◽  
Drug Metabolizing Enzymes ◽  
Metabolizing Enzymes ◽  
The Impact

scholarly journals Influence ofβS-Globin Haplotypes and Hydroxyurea on Arginase I Levels in Sickle Cell Disease

2016 ◽  
Vol 2016 ◽  
pp. 1-5 ◽  
Author(s):  
J. A. Moreira ◽  
R. P. G. Machado ◽  
M. R. Laurentino ◽  
Romelia Pinheiro Gonçalves Lemes ◽  
M. C. Barbosa ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Clinical Symptoms ◽  
Inverse Correlation ◽  
Control Group ◽  
Cell Disease ◽  
Hemoglobin F ◽  
Significance Level ◽  
The Impact ◽  
Arginase I

Introduction.Sickle cell disease (SCD) is characterized by hemoglobin S homozygosity, leading to hemolysis and vasoocclusion. The hemolysis releases arginase I, an enzyme that decreases the bioavailability of nitric oxide, worsening the symptoms. The different SCD haplotypes are related to clinical symptoms and varied hemoglobin F (HbF) concentration. The aim of this study was to evaluate the impact of theβS gene haplotypes and HbF concentration on arginase I levels in SCD patients.Methods.Fifty SCD adult patients were enrolled in the study and 20 blood donors composed the control group. Arginase I was measured by ELISA. TheβS haplotypes were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Statistical analyses were performed with GraphPad Prism program and the significance level wasp<0.05.Results.Significant increase was observed in the arginase I levels in SCD patients compared to the control group (p<0.0001). The comparison between the levels of arginase I in three haplotypes groups showed a difference between the Bantu/Bantu×Bantu/Benin groups; Bantu/Bantu×Benin/Benin, independent of HU dosage. An inverse correlation with the arginase I levels and HbF concentration was observed.Conclusion.The results support the hypothesis that arginase I is associated with HbF concentration, also measured indirectly by the association with haplotypes.

scholarly journals Effects of Synergistic Massage and Physical Exercise on the Expression of Angiogenic Markers in Rat Tendons

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Waldemar Andrzejewski ◽  
Krzysztof Kassolik ◽  
Piotr Dziegiel ◽  
Bartosz Pula ◽  
Katarzyna Ratajczak-Wielgomas ◽  
...  
Keyword(s):  
Physical Exercise ◽  
Mrna Level ◽  
Molecular Techniques ◽  
Control Group ◽  
Key Factors ◽  
Entire Study ◽  
The Third ◽  
Physical Exercises ◽  
Varied Duration ◽  
The Impact

Physical exercise and massage are regarded as key factors in regulating tendon structure. However, information on the mechanism through which massage influences the structure and biology of a tendon is scarce. In this study, we attempted to define the impact of these two activities on rat tendons by using morphological and molecular techniques, determining the expression of VEGF-A, FGF-2, and CD34 in the tendons of rats subjected to 10 weeks of physical exercise (running) with massage of varied duration. The group of rats that was trained and massaged during the entire study was characterized by the highest expression of these markers, compared to the rats subjected to massage before training and to the control group subjected to physical exercises only. The greatest significant differences, compared to the control, were noted in the expression of all the studied markers at mRNA level, and in the case of VEGF-A, at protein level, in the third and fifth weeks of the experiment. The results of this study could point to the synergistic impact of simultaneous massage and physical exercise on the expression of angiogenesis markers in rat tendons.

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