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1.
Int J Mol Sci ; 24(16)2023 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-37628918

RESUMO

Previously, we demonstrated in pigs that renal denervation halves glucose release during hypoglycaemia and that a prenatal dexamethasone injection caused increased ACTH and cortisol concentrations as markers of a heightened hypothalamic pituitary adrenal axis (HPAA) during hypoglycaemia. In this study, we investigated the influence of an altered HPAA on renal glucose release during hypoglycaemia. Pigs whose mothers had received two late-gestational dexamethasone injections were subjected to a 75 min hyperinsulinaemic-hypoglycaemic clamp (<3 mmol/L) after unilateral surgical denervation. Para-aminohippurate (PAH) clearance, inulin, sodium excretion and arterio-venous blood glucose difference were measured every fifteen minutes. The statistical analysis was performed with a Wilcoxon signed-rank test. PAH, inulin, the calculated glomerular filtration rate and plasma flow did not change through renal denervation. Urinary sodium excretion increased significantly (p = 0.019). Side-dependent renal net glucose release (SGN) decreased by 25 ± 23% (p = 0.004). At 25 percent, the SGN decrease was only half of that observed in non-HPAA-altered animals in our prior investigation. The current findings may suggest that specimens with an elevated HPAA undergo long-term adaptations to maintain glucose homeostasis. Nonetheless, the decrease in SGN warrants further investigations and potentially caution in performing renal denervation in certain patient groups, such as diabetics at risk of hypoglycaemia.


Assuntos
Hipoglicemia , Hipoglicemiantes , Feminino , Animais , Suínos , Gravidez , Glucose , Sistema Hipotálamo-Hipofisário , Inulina , Sistema Hipófise-Suprarrenal , Ácido p-Aminoipúrico , Dexametasona/efeitos adversos , Denervação
2.
Drug Metab Dispos ; 49(7): 521-529, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33941544

RESUMO

In traditional pharmacokinetic models, blood flow or liquid transit is often expressed as first-order kinetics. When the flow expression by first-order kinetics is used for dynamic simulation, the flow velocity illogically depends on the step size of a solver of ordinary differential equations. In this study, we propose flow modeling using hybrid automata that combine ordinary differential equations and recursive equations, and we have preliminarily applied the constructed models to several examples. The blood concentration-time profiles of p-aminohippurate and propranolol after intravenous administration were successfully reproduced by simple hybrid automata. The simulation results of one-dimensional tube flow have demonstrated that the fluid velocity in the hybrid automata was independent of the step size of the ordinary differential equation solver. A body fluid model coordinated various flows in a human body with scheduled daily activities and could be used as a drug container to describe formulation-dependent disposition of 5-aminosalicylic acid and enterohepatic circulation of a virtual drug. These findings suggested that flow modeling using hybrid automata could avoid the logical inconsistency in the traditional pharmacokinetic modeling and that the hybrid automata have high versatility and a wide range of applicability to pharmacokinetic analysis. Because our method can define various intervals for multiple recursive equations, the resolution of a specific part of a model can be adjusted relatively freely while the whole body is being roughly modeled, which would be beneficial to refine a coarse model into a fine model in future. SIGNIFICANCE STATEMENT: There is a logical inconsistency in flow expression by first-order kinetics in ordinary differential equations used in traditional pharmacokinetic modeling. It is difficult to model a whole human body using flow models in partial differential equations because of the excessive calculation costs. Our simulations on tube flow and body fluids have demonstrated that the flow modeling using hybrid automata could avoid the problems. The preliminary applications of hybrid automata to several examples highlighted its high versatility in pharmacokinetic analysis.


Assuntos
Modelos Cardiovasculares , Administração Intravenosa , Velocidade do Fluxo Sanguíneo , Simulação por Computador , Humanos , Propranolol/administração & dosagem , Propranolol/farmacocinética , Ácido p-Aminoipúrico/administração & dosagem , Ácido p-Aminoipúrico/farmacocinética
3.
CPT Pharmacometrics Syst Pharmacol ; 9(10): 571-583, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32977369

RESUMO

Chronic kidney disease (CKD) has significant effects on renal clearance (CLr ) of drugs. Physiologically-based pharmacokinetic (PBPK) models have been used to predict CKD effects on transporter-mediated renal active secretion and CLr for hydrophilic nonpermeable compounds. However, no studies have shown systematic PBPK modeling of renal passive reabsorption or CLr for hydrophobic permeable drugs in CKD. The goal of this study was to expand our previously developed and verified mechanistic kidney model to develop a universal model to predict changes in CLr in CKD for permeable and nonpermeable drugs that accounts for the dramatic nonlinear effect of CKD on renal passive reabsorption of permeable drugs. The developed model incorporates physiologically-based tubular changes of reduced water reabsorption/increased tubular flow rate per remaining functional nephron in CKD. The final adaptive kidney model successfully (absolute fold error (AFE) all < 2) predicted renal passive reabsorption and CLr for 20 permeable and nonpermeable test compounds across the stages of CKD. In contrast, use of proportional glomerular filtration rate reduction approach without addressing tubular adaptation processes in CKD to predict CLr generated unacceptable CLr predictions (AFE = 2.61-7.35) for permeable compounds in severe CKD. Finally, the adaptive kidney model accurately predicted CLr of para-amino-hippuric acid and memantine, two secreted compounds, in CKD, suggesting successful integration of active secretion into the model, along with passive reabsorption. In conclusion, the developed adaptive kidney model enables mechanistic predictions of in vivo CLr through CKD progression without any empirical scaling factors and can be used for CLr predictions prior to assessment of drug disposition in renal impairment.


Assuntos
Rim/metabolismo , Taxa de Depuração Metabólica/efeitos dos fármacos , Insuficiência Renal Crônica/sangue , Reabsorção Renal/efeitos dos fármacos , Transporte Biológico , Progressão da Doença , Taxa de Filtração Glomerular/efeitos dos fármacos , Humanos , Rim/fisiologia , Memantina/metabolismo , Modelos Biológicos , Farmacocinética , Valor Preditivo dos Testes , Insuficiência Renal/metabolismo , Insuficiência Renal Crônica/classificação , Insuficiência Renal Crônica/diagnóstico , Insuficiência Renal Crônica/fisiopatologia , Ácido p-Aminoipúrico/metabolismo
4.
Diabetes ; 69(12): 2700-2708, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32737116

RESUMO

The objective of this study was to compare the ratio of renal oxygen availability (RO2) to glomerular filtration rate (GFR), a measure of relative renal hypoxia, in adolescents with and without type 1 diabetes (T1D) and relate the ratio to albuminuria, renal plasma flow (RPF), fat mass, and insulin sensitivity (M/I). RO2 was estimated by blood oxygen level-dependent MRI; fat mass was estimated by DXA; GFR and RPF were estimated by iohexol and p-aminohippurate clearance; albuminuria was estimated by urine albumin-to-creatinine ratio (UACR); and M/I was estimated from steady-state glucose infusion rate/insulin (mg/kg/min) by hyperglycemic clamp in 50 adolescents with T1D (age 16.1 ± 3.0 years, HbA1c 8.6 ± 1.2%) and 20 control patients of similar BMI (age 16.1 ± 2.9 years, HbA1c 5.2 ± 0.2%). The RO2:GFR (ms/mL/min) was calculated as RO2 (T2*, ms) divided by GFR (mL/min). Whole-kidney RO2:GFR was 25% lower in adolescents with T1D versus control patients (P < 0.0001). In adolescents with T1D, lower whole-kidney RO2:GFR was associated with higher UACR (r = -0.31, P = 0.03), RPF (r = -0.52, P = 0.0009), and fat mass (r = -0.33, P = 0.02). Lower medullary RO2:GFR was associated with lower M/I (r = 0.31, P = 0.03). In conclusion, adolescents with T1D exhibited relative renal hypoxia that was associated with albuminuria and with increased RPF, fat mass, and insulin resistance. These data suggest a potential role of renal hypoxia in the development of diabetic kidney disease.


Assuntos
Diabetes Mellitus Tipo 1/complicações , Hipóxia , Oxigênio/metabolismo , Adiposidade , Adolescente , Composição Corporal , Criança , Diabetes Mellitus Tipo 1/metabolismo , Nefropatias Diabéticas , Feminino , Furosemida , Taxa de Filtração Glomerular , Técnica Clamp de Glucose , Hemoglobinas Glicadas/genética , Hemoglobinas Glicadas/metabolismo , Humanos , Insulina , Iohexol/farmacologia , Masculino , Adulto Jovem , Ácido p-Aminoipúrico/farmacologia
5.
Am J Physiol Gastrointest Liver Physiol ; 319(2): G133-G141, 2020 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-32538141

RESUMO

Xenometabolites from microbial and plant sources are thought to confer beneficial as well as deleterious effects on host physiology. Studies determining absorption and tissue uptake of xenometabolites are limited. We utilized a conscious catheterized pig model to evaluate interorgan flux of annotated known and suspected xenometabolites, derivatives, and bile acids. Female pigs (n = 12, 2-3 mo old, 25.6 ± 2.2 kg) had surgically implanted catheters across portal-drained viscera (PDV), splanchnic compartment (SPL), liver, kidney, and hindquarter muscle. Overnight-fasted arterial and venous plasma was collected simultaneously in a conscious state and stored at -80°C. Thawed samples were analyzed by liquid chromatography-mass spectrometry. Plasma flow was determined with para-aminohippuric acid dilution technology and used to calculate net organ balance for each metabolite. Significant organ uptake or release was determined if net balance differed from zero. A total of 48 metabolites were identified in plasma, and 31 of these had at least one tissue with a significant net release or uptake. All bile acids, indole-3-acetic acid, indole-3-arylic acid, and hydrocinnamic acid were released from the intestine and taken up by the liver. Indole-3-carboxaldehyde, p-cresol glucuronide, 4-hydroxyphenyllactic acid, dodecanendioic acid, and phenylacetylglycine were also released from the intestines. Liver or kidney uptake was noted for indole-3-acetylglycine, p-cresol glucuronide, atrolactic acid, and dodecanedioic acid. Indole-3-carboxaldehyde, atrolactic acid, and dodecanedioic acids showed net release from skeletal muscle. The results confirm gastrointestinal origins for several known xenometabolites in an in vivo overnight-fasted conscious pig model, whereas nongut net release of other putative xenometabolites suggests a more complex metabolism.NEW & NOTEWORTHY Xenometabolites from microbe origins influence host health and disease, but absorption and tissue uptake of these metabolites remain speculative. Results herein are the first to demonstrate in vivo organ uptake and release of these metabolites. We used a conscious catheterized pig model to confirm gastrointestinal origins for several xenometabolites (e.g., indolic compounds, 4-hydroxyphenyllactic acid, dodecanendioic acid, and phenylacetylgycine). Liver and kidney were major sites for xenometabolite uptake, likely highlighting liver conjugation metabolism and renal excretion.


Assuntos
Intestinos/fisiologia , Rim/fisiologia , Fígado/metabolismo , Músculo Esquelético/fisiologia , Ácido p-Aminoipúrico/farmacocinética , Animais , Transporte Biológico , Feminino , Fenóis/sangue , Fenóis/metabolismo , Sistema Porta , Suínos , Ácido p-Aminoipúrico/sangue
6.
Mol Pharmacol ; 97(6): 384-391, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32234809

RESUMO

Organic anion transporter 1 (OAT1), expressed at the basolateral membrane of renal proximal tubule epithelial cells, mediates the renal excretion of many clinically important drugs. Previous study in our laboratory demonstrated that ubiquitin conjugation to OAT1 leads to OAT1 internalization from the cell surface and subsequent degradation. The current study showed that the ubiquitinated OAT1 accumulated in the presence of the proteasomal inhibitors MG132 and ALLN rather than the lysosomal inhibitors leupeptin and pepstatin A, suggesting that ubiquitinated OAT1 degrades through proteasomes. Anticancer drugs bortezomib and carfilzomib target the ubiquitin-proteasome pathway. We therefore investigate the roles of bortezomib and carfilzomib in reversing the ubiquitination-induced downregulation of OAT1 expression and transport activity. We showed that bortezomib and carfilzomib extremely increased the ubiquitinated OAT1, which correlated well with an enhanced OAT1-mediated transport of p-aminohippuric acid and an enhanced OAT1 surface expression. The augmented OAT1 expression and transport activity after the treatment with bortezomib and carfilzomib resulted from a reduced rate of OAT1 degradation. Consistent with this, we found decreased 20S proteasomal activity in cells that were exposed to bortezomib and carfilzomib. In conclusion, this study identified the pathway in which ubiquitinated OAT1 degrades and unveiled a novel role of anticancer drugs bortezomib and carfilzomib in their regulation of OAT1 expression and transport activity. SIGNIFICANCE STATEMENT: Bortezomib and carfilzomib are two Food and Drug Administration-approved anticancer drugs, and proteasome is the drug target. In this study, we unveiled a new role of bortezomib and carfilzomib in enhancing OAT1 expression and transport activity by preventing the degradation of ubiquitinated OAT1 in proteasomes. This finding provides a new strategy in regulating OAT1 function that can be used to accelerate the clearance of drugs, metabolites, or toxins and reverse the decreased expression under disease conditions.


Assuntos
Antineoplásicos/farmacologia , Transporte Biológico Ativo/efeitos dos fármacos , Bortezomib/farmacologia , Oligopeptídeos/farmacologia , Proteína 1 Transportadora de Ânions Orgânicos/metabolismo , Complexo de Endopeptidases do Proteassoma/metabolismo , Inibidores de Proteassoma/farmacologia , Células HEK293 , Humanos , Leupeptinas/farmacologia , Proteólise , Ubiquitinação , Ácido p-Aminoipúrico/metabolismo
7.
Spectrochim Acta A Mol Biomol Spectrosc ; 228: 117805, 2020 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-31787536

RESUMO

Herein, we report a method for the synthesis of 4-aminohippuric acid (4-AHA) reduced/stabilized gold nanoparticles and their peroxidase mimicking properties for the colorimetric detection of Fe3+ and Hg2+. The synthesis of nanoparticles was evidenced by appearance of bright red color and an absorption peak at 518 nm. Transmission electron microscopic (TEM) characterization revealed the nanoparticles to be spherical with average size of about 5.9 ± 1.7 nm. X-ray diffraction (XRD) analysis established highly crystalline nature of the nanoparticles. The synthesized nanoparticles have shown very good peroxidase mimicking property; exhibiting the catalytic oxidation of the chromogen 3,3',5,5'-tetramethyl benzidine (TMB) to a blue color product, in the presence of hydrogen peroxide. The peroxidase mimicking activity of the nanoparticles was found to be selectivity enhanced in the presence of Fe3+ and Hg2+ while there was no change in the activity in the presence of other concomitant ions. The mechanism studies revealed that the synthesized gold nanoparticles assisted in electron transfer during the catalytic process however the stimulation of peroxidase-like activity in the presence of Fe3+ and Hg2+ is owed to both generation of hydroxyl radical and accelerated electron transfer. The assay was made selective for iron by the addition of cysteine in acetate buffer; whereas the selective detection of mercury was achieved by carrying out the assay in citrate buffer. The linear ranges for the determination of Fe3+ and Hg2+ in deionized water were found to be: 5-50 ppb and 5-200 ppb respectively. The limits of quantification (LOQ) for Fe3+ and Hg2+ were 4.0 and 2.5 ppb respectively. The assay was applied for the determination of Fe3+ and Hg2+ in drinking and ground water samples. The method holds potential for the on-field screening of these metal ions in real environmental samples.


Assuntos
Ouro/química , Água Subterrânea/análise , Ferro/análise , Mercúrio/análise , Nanopartículas Metálicas/química , Peroxidase/química , Ácido p-Aminoipúrico/química , Colorimetria
8.
Artigo em Inglês | MEDLINE | ID: mdl-31004849

RESUMO

In order to study the renal function, in terms of glomerular filtration and effective renal plasma flow, in broiler chickens and pigs, an ultra-high performance liquid chromatography-tandem mass spectrometry method for the simultaneous determination of iohexol, p-aminohippuric acid (PAH) and exogenously administered creatinine in plasma was developed and validated. Sample preparation consisted of a deproteinization step using methanol for porcine plasma and an Ostro™ Protein Precipitation & Phospholipid Removal Plate was used for broiler chicken plasma. Chromatographic separation was achieved on a Hypersil Gold aQ column using 0.1% formic acid in water and 0.1% formic acid in methanol as mobile phases. The total run time was limited to 10 min. Matrix-matched calibration curves for iohexol and PAH were prepared and good linearity (r ≥ 0.9973; gof ≤ 6.17%) was achieved over the concentration range tested (0.25-90 µg/mL). Limits of quantification were 0.25 µg/mL for iohexol and PAH. Water was used as surrogate matrix for analysis of creatinine in plasma. This surrogate calibration curve showed good linearity over the concentration range tested (0.25-90 µg/mL) (r ≥ 0.9979; gof ≤ 5.66%). For creatinine, the relative lower limit of quantification was 201.03 ±â€¯49.20% and 60.14 ±â€¯7.64% for chicken and porcine plasma, respectively. The results for within-day and between-day precision and accuracy fell within the specified ranges. This straightforward, cost-effective and rapid method, determining iohexol, PAH and creatinine within one single chromatographic run, has been successfully used for the analysis in porcine and broiler chicken plasma samples in order to determine the renal function of these species.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Creatinina/sangue , Iohexol/análise , Espectrometria de Massas em Tandem/métodos , Ácido p-Aminoipúrico/sangue , Animais , Galinhas , Creatinina/farmacocinética , Iohexol/farmacocinética , Testes de Função Renal , Limite de Detecção , Modelos Lineares , Reprodutibilidade dos Testes , Suínos , Ácido p-Aminoipúrico/farmacocinética
9.
J Asian Nat Prod Res ; 21(8): 754-771, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30606060

RESUMO

Andrographolide, a major bioactive compound isolated from Andrographis paniculata (Burm. F.) Nees, was evaluated for its effects on the hOAT1 membrane transporter. Substrate determination and inhibition of hOAT1-mediated uptake transport assay was carried out using recombinant CHO-hOAT1 cells. The results showed that the uptake ratio of andrographolide was less than 2.0 at all concentrations tested, indicating that andrographolide is not a hOAT1 substrate. Andrographolide has no significant effects on the p-aminohippuric acid uptake and on the mRNA and protein expression of hOAT1. In conclusion, andrographolide may not pose a drug-herb interaction risk related to hOAT1.


Assuntos
Diterpenos/farmacologia , Proteína 1 Transportadora de Ânions Orgânicos/antagonistas & inibidores , Animais , Células CHO , Proliferação de Células/efeitos dos fármacos , Cricetulus , Diterpenos/farmacocinética , Interações Ervas-Drogas , Humanos , Simulação de Acoplamento Molecular , Proteína 1 Transportadora de Ânions Orgânicos/análise , Proteína 1 Transportadora de Ânions Orgânicos/química , Proteína 1 Transportadora de Ânions Orgânicos/genética , Probenecid/química , Probenecid/farmacologia , Ácido p-Aminoipúrico/farmacocinética
10.
Br J Clin Pharmacol ; 85(5): 900-907, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30666700

RESUMO

AIMS: Reduced nitric oxide (NO) availability may adversely affect renal perfusion and glomerular filtration. The aim of the present study was to characterize in detail the pharmacological effects of VAS203, an inhibitor of NO synthase, on renal haemodynamics in humans. METHODS: This double-blind, randomized, placebo-controlled, cross-over phase-I-study comprised 18 healthy men. Renal haemodynamics were assessed with constant-infusion input-clearance technique with p-aminohippurate and inulin for renal plasma flow (RPF) and glomerular filtration rate (GFR), respectively. After baseline measurement, a constant infusion of the tetrahydrobiopterin analogue ronopterin (VAS203, total 10 mg/kg body weight) or placebo was administered at random order for 6 hours additionally. After a wash-out phase of 28 days, the second course was applied. In parallel, markers of early kidney injury and renal function were assessed repeatedly up to 48 hours after starting VAS203/placebo-infusion. RESULTS: VAS203-infusion resulted in a significant decrease of RPF (P < .0001) and GFR (P < .001) compared to placebo, but magnitude was within the physiological range. RPF and GFR recovered partly 2 hours after end of VAS203-infusion and was normal at beginning of the second infusion period. Compared to placebo, preglomerular resistance (P < .0001), and to lesser extent postglomerular resistance (P < .0001) increased, resulting in a decrease of intraglomerular pressure (P < .01). No treatment related effect on markers of early kidney injury, and on renal function (P for all >.20) have been observed. CONCLUSIONS: Our phase-I-study in healthy humans indicates that VAS203 (10 mg/kg body weight) reduces renal perfusion and glomerular function within the physiological range mainly due to vasoconstriction at the preglomerular site.


Assuntos
Injúria Renal Aguda/induzido quimicamente , Glomérulos Renais/efeitos dos fármacos , Eliminação Renal/efeitos dos fármacos , Injúria Renal Aguda/diagnóstico , Adulto , /efeitos adversos , Estudos Cross-Over , Método Duplo-Cego , Voluntários Saudáveis , Humanos , Infusões Intravenosas , Inulina/administração & dosagem , Inulina/metabolismo , Glomérulos Renais/irrigação sanguínea , Glomérulos Renais/fisiologia , Masculino , Fluxo Sanguíneo Regional/efeitos dos fármacos , Vasoconstrição/efeitos dos fármacos , Adulto Jovem , Ácido p-Aminoipúrico/administração & dosagem , Ácido p-Aminoipúrico/metabolismo
11.
J Am Heart Assoc ; 7(16): e008455, 2018 08 21.
Artigo em Inglês | MEDLINE | ID: mdl-30369310

RESUMO

Background The management of the cardiorenal syndrome in advanced heart failure is challenging, and the role of inotropic drugs has not been fully defined. Our aim was to compare the renal effects of levosimendan versus dobutamine in patients with heart failure and renal impairment. Methods and Results In a randomized double-blind study, we assigned patients with chronic heart failure (left ventricular ejection fraction <40%) and impaired renal function (glomerular filtration rate <80 mL/min per 1.73 m2) to receive either levosimendan (loading dose 12 µg/kg+0.1 µg/kg per minute) or dobutamine (7.5  µg/kg per minute) for 75 minutes. A pulmonary artery catheter was used for measurements of systemic hemodynamics, and a renal vein catheter was used to measure renal plasma flow by the infusion clearance technique for PAH (para-aminohippurate) corrected by renal extraction of PAH . Filtration fraction was measured by renal extraction of chromium ethylenediamine tetraacetic acid. A total of 32 patients completed the study. Following treatment, the levosimendan and dobutamine groups displayed similar increases in renal blood flow (22% and 26%, respectively) with no significant differences between groups. Glomerular filtration rate increased by 22% in the levosimendan group but remained unchanged in the dobutamine group ( P=0.012). Filtration fraction was not affected by levosimendan but decreased by 17% with dobutamine ( P=0.045). Conclusions In patients with chronic heart failure and renal impairment, levosimendan increases glomerular filtration rate to a greater extent than dobutamine and thus may be the preferred inotropic agent for treating patients with the cardiorenal syndrome. Clinical Trial Registration URL: https://www.clinicaltrials.gov . Unique identifier: NCT 02133105.


Assuntos
Síndrome Cardiorrenal/tratamento farmacológico , Cardiotônicos/uso terapêutico , Dobutamina/uso terapêutico , Insuficiência Cardíaca/tratamento farmacológico , Insuficiência Renal/metabolismo , Simendana/uso terapêutico , Idoso , Método Duplo-Cego , Feminino , Taxa de Filtração Glomerular , Insuficiência Cardíaca/complicações , Monitorização Hemodinâmica , Hemodinâmica , Humanos , Masculino , Pessoa de Meia-Idade , Circulação Renal , Insuficiência Renal/complicações , Fluxo Plasmático Renal , Ácido p-Aminoipúrico
12.
J Appl Toxicol ; 38(7): 968-977, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29460972

RESUMO

The kidney plays a major part in the elimination of many drugs and their metabolites, and drug-induced kidney injury commonly alters either glomerular filtration or tubular transport, or both. However, the renal excretion pathway of drugs has not been fully elucidated at different stages of renal injury. This study aimed to evaluate the alteration of renal excretion pathways in gentamicin (GEN)-induced renal injury in rats. Results showed that serum cystatin C, creatinine and urea nitrogen levels were greatly increased by the exposure of GEN (100 mg kg-1 ), and creatinine concentration was increased by 39.7% by GEN (50 mg kg-1 ). GEN dose-dependently upregulated the protein expression of rOCT1, downregulated rOCT2 and rOAT1, but not affected rOAT2. Efflux transporters, rMRP2, rMRP4 and rBCRP expressions were significantly increased by GEN(100), and the rMATE1 level was markedly increased by GEN(50) but decreased by GEN(100). GEN(50) did not alter the urinary excretion of inulin, but increased metformin and furosemide excretion. However, GEN(100) resulted in a significant decrease of the urinary excretion of inulin, metformin and p-aminohippurate. In addition, urinary metformin excretions in vivo were significantly decreased by GEN(100), but slightly increased by GEN(50). These results suggested that GEN(50) resulted in the induction of rOCTs-rMATE1 and rOAT3-rMRPs pathway, but not changed the glomerular filtration rate, and GEN(100)-induced acute kidney injury caused the downregulated function of glomerular filtration -rOCTs-rMATE1 and -rOAT1-rMRPs pathway.


Assuntos
Injúria Renal Aguda/metabolismo , Gentamicinas , Rim/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Eliminação Renal , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Injúria Renal Aguda/induzido quimicamente , Injúria Renal Aguda/fisiopatologia , Animais , Antiporters/metabolismo , Modelos Animais de Doenças , Furosemida/metabolismo , Taxa de Filtração Glomerular , Inulina/urina , Rim/fisiopatologia , Masculino , Metformina/farmacocinética , Metformina/urina , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , Proteínas de Transporte de Cátions Orgânicos/metabolismo , Ratos Wistar , Ácido p-Aminoipúrico/metabolismo
13.
Pharmacology ; 101(3-4): 176-183, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29353278

RESUMO

Lansoprazole, a proton pump inhibitor, potently inhibits human organic anion transporter, hOAT3 (SLC22A8). Lansoprazole has an asymmetric atom in its structure and is clinically administered as a racemic mixture of (R)-and (S)-enantiomers. However, little is known about the stereoselective inhibitory potencies of lansoprazole against hOAT3 and its homolog, hOAT1. In the present study, the stereoselective inhibitory effect of lansoprazole was evaluated using hOAT1-and hOAT3-expressing cultured cells. hOAT1 and hOAT3 transported [14C]p-aminohippurate and [3H]estrone-3-sulfate (ES) with Michaelis-Menten constants of 29.8 ± 4.0 and 30.1 ± 9.0 µmol/L respectively. Lansoprazole enantiomers inhibited hOAT1- and hOAT3-mediated transport of each substrate in a concentration-dependent manner. The IC50 value of (S)-lansoprazole against hOAT3-mediated transport of [3H]ES (0.61 ± 0.08 µmol/L) was significantly lower than that of (R)-lansoprazole (1.75 ± 0.31 µmol/L). In contrast, stereoselectivity was not demonstrated for the inhibition of hOAT1. Furthermore, (S)-lansoprazole inhibited hOAT3-mediated transport of pemetrexed and methotrexate (hOAT3 substrates) more strongly than the corresponding (R)-lansoprazole. This study is the first to demonstrate that the stereoselective inhibitory potency of (S)-lansoprazole against hOAT3 is greater than that of (R)-lansoprazole. The present findings provide novel information about the drug interactions associated with lansoprazole.


Assuntos
Lansoprazol/química , Lansoprazol/farmacologia , Transportadores de Ânions Orgânicos Sódio-Independentes/antagonistas & inibidores , Inibidores da Bomba de Prótons/química , Inibidores da Bomba de Prótons/farmacologia , Interações Medicamentosas , Estrona/análogos & derivados , Estrona/farmacologia , Células HEK293 , Humanos , Rim/metabolismo , Metotrexato/farmacologia , Proteína 1 Transportadora de Ânions Orgânicos/antagonistas & inibidores , Proteína 1 Transportadora de Ânions Orgânicos/genética , Proteína 1 Transportadora de Ânions Orgânicos/metabolismo , Transportadores de Ânions Orgânicos Sódio-Independentes/genética , Transportadores de Ânions Orgânicos Sódio-Independentes/metabolismo , Pemetrexede/farmacologia , Probenecid/farmacologia , Estereoisomerismo , Ácido p-Aminoipúrico/farmacologia
14.
Int J Pharm ; 537(1-2): 172-182, 2018 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-29277663

RESUMO

This study aimed to demonstrate that organic anion transporters (OATs) mediate the drug-drug interaction (DDI) between piperacillin and tazobactam. After co-administration with piperacillin in rats, the AUC of tazobactam in plasma was significantly increased, and t1/2ß was prolonged with significant reduction in plasma clearance, renal clearance and cumulative urinary excretion. In rat and human kidney slices, probenecid, p-aminohippurate and benzylpenicillin inhibited the uptake of piperacillin and tazobactam. Piperacillin significantly inhibited the uptake of tazobactam. Moreover, the uptakes of piperacillin, tazobactam and sulbactam in hOAT1/3-HEK293 cells were significantly higher compared with mock-HEK293 cells, respectively. Piperacillin significantly inhibited the uptake of tazobactam in hOAT1/3-HEK293 cells. The Km values of tazobactam (431 ±â€¯67 µM for hOAT1, 377 ±â€¯63 µM for hOAT3) were significantly higher than those of piperacillin (37 ±â€¯5 µM for hOAT1, 172 ±â€¯28 µM for hOAT3). This suggested that piperacillin has a stronger affinity to hOAT1/3 than tazobactam. Meanwhile, the Km values of tazobactam were increased in the presence of piperacillin with unchanged Vmax. This indicated that piperacillin inhibited the uptake of tazobactam in a competitive manner. In conclusion, piperacillin and tazobactam are the substrates of hOAT1/3, and OAT1/3 mediate the DDI between piperacillin and tazobactam.


Assuntos
Interações Medicamentosas/fisiologia , Proteína 1 Transportadora de Ânions Orgânicos/metabolismo , Transportadores de Ânions Orgânicos Sódio-Independentes/metabolismo , Ácido Penicilânico/análogos & derivados , Piperacilina/farmacologia , Animais , Linhagem Celular , Células HEK293 , Humanos , Rim/efeitos dos fármacos , Rim/metabolismo , Masculino , Ácido Penicilânico/farmacologia , Penicilina G/metabolismo , Probenecid/metabolismo , Ratos , Ratos Wistar , Sulbactam/farmacologia , Tazobactam , Ácido p-Aminoipúrico/metabolismo
15.
Animal ; 12(2): 232-238, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28712380

RESUMO

In nutrition studies, para-aminohippuric acid (PAH) is a marker frequently used to measure blood flow in pigs, which is essential for estimating portal-drained viscera (PDV) flux of nutrients. The aim of this study was to evaluate the PAH analytical method by means of qualimetric statistical procedures to estimate the matrix effect and the accuracy and limits of quantitation of the method. Net PDV flux of nutrients was determined in five multi-catheterized pigs using water, plasma or commercial serum as standard matrix. A proportional systematic error due to matrix effect was found for plasma and serum. Mean recovery was 99.4%, and intra- and inter-day precision of the method was 2.4% and 3.8% relative standard deviation, respectively. The limit of quantification was 0.22 mg PAH/l. Use of water for the PAH standard curves underestimated portal blood flow compared with PAH standards prepared with plasma or commercial serum (706, 954 and 927 ml/min; P<0.05, respectively). Consequently, PDV O2 consumption, glucose and amino acids fluxes were underestimated by 33% (P<0.001). In conclusion, our results stress the importance of using plasma from pigs not infused with PAH or alternatively commercial pig serum to prepare PAH standards to determine blood flow in pigs to avoid underestimation of blood flow.


Assuntos
Estudos de Avaliação como Assunto , Suínos/metabolismo , Ácido p-Aminoipúrico/análise , Aminoácidos/metabolismo , Animais , Calibragem , Glucose/metabolismo , Indicadores e Reagentes/análise , Fígado/metabolismo , Oxigênio/metabolismo , Consumo de Oxigênio , Veia Porta/metabolismo , Suínos/sangue , Vísceras/metabolismo
16.
Nephron ; 136(2): 151-157, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28329736

RESUMO

BACKGROUND/AIMS: Little information is available about the tubular functions and the renal adjustments that take place in obese subjects after a protein meal. How the excess fat may affect renal response to dietary proteins is currently only partially understood. This paper aims to address (i) whether severe obesity, in the absence of other comorbidities, is responsible of kidney dysfunction at either the glomerular or the tubular level and (ii) whether it compromises renal adaptations to a large protein meal. METHODS: Twenty-eight obese subjects without albuminuria, along with 20 control subjects, age and gender matched, have been studied. The glomerular filtration rate (GFR; inulin clearance), renal plasma flow (p-aminohippurate clearance), the proximal tubular function (lithium clearance), the fractional excretion of sodium (FPRNa) have been measured at the basal level (steady state) and after a protein meal (perturbation). RESULTS: Under steady state conditions, filtration fraction, proximal tubular sodium handling and the FPRNa were not significantly different in non proteinuric obese subjects compared with controls. However, a protein meal led to a delayed glomerular hyperfiltration in obese patients compared with controls. CONCLUSION: This study shows that obese patients, in the absence of significant comorbidities, have a normal proximal tubule Na+ absorption at basal; conversely, these subjects showed a different response to a protein meal compared with normal subjects in terms of changes of GFR. Overall, these results suggest that the modified hemodynamic response to a protein meal might be the earliest hallmark of future kidney dysfunction in obese subjects.


Assuntos
Nefropatias/fisiopatologia , Refeições , Carne/efeitos adversos , Obesidade Mórbida/fisiopatologia , Circulação Renal , Adulto , Albuminúria , Proteínas na Dieta/efeitos adversos , Feminino , Taxa de Filtração Glomerular , Hemodinâmica , Humanos , Testes de Função Renal , Túbulos Renais Proximais/fisiopatologia , Masculino , Pessoa de Meia-Idade , Ácido p-Aminoipúrico/urina
17.
Am J Physiol Renal Physiol ; 312(6): F992-F997, 2017 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-28249837

RESUMO

Hyperuricemia has been reported to affect renal hemodynamics. In a recent study, both low and high levels of serum uric acid (SUA) were found to be associated with loss of kidney function. The goal of this study was to evaluate the relationship between SUA levels and intrarenal hemodynamic parameters in healthy subjects, using plasma clearance of para-aminohippurate (CPAH) and inulin (Cin). Renal and glomerular hemodynamics were evaluated by simultaneous measurements of CPAH and Cin in 48 healthy subjects (54.6 ± 13.4 yr). Intrarenal hemodynamic parameters, including efferent and afferent (Ra) arteriolar resistance, were calculated using Gómez's formulas. Relationships of SUA levels with these intrarenal hemodynamic parameters were examined. In quadratic regression analysis, SUA levels had a significant inverse U-shaped relationship with Cin (P < 0.0001, R2 = 0.350) and CPAH (P = 0.0093, R2 = 0.188) and a U-shaped relationship with Ra (P = 0.0011, R2 = 0.262). In multiple regression analysis with normal (3.5-6.0 mg/dl) and mildly low or high (<3.5 or >6.0 mg/dl) SUA levels entered as dummy variables of zero and one, respectively, mildly low or high SUA levels were significantly and independently associated with Ra (ß = 0.230, P = 0.0403) after adjustment for several factors (R2 = 0.597, P < 0.0001). Both mild hyperuricemia and mild hypouricemia are significantly associated with increased Ra, although weakly. The increase in Ra in subjects with mild hyperuricemia or hypouricemia may be related to renal hemodynamic abnormalities, possibly leading to a decline in renal function.


Assuntos
Hemodinâmica , Hiperuricemia/sangue , Hiperuricemia/fisiopatologia , Rim/irrigação sanguínea , Circulação Renal , Ácido Úrico/sangue , Adulto , Idoso , Biomarcadores/sangue , Feminino , Taxa de Filtração Glomerular , Voluntários Saudáveis , Humanos , Infusões Intravenosas , Inulina/administração & dosagem , Inulina/metabolismo , Masculino , Pessoa de Meia-Idade , Modelos Biológicos , Análise de Regressão , Fluxo Plasmático Renal , Ácido p-Aminoipúrico/administração & dosagem , Ácido p-Aminoipúrico/metabolismo
18.
Anal Biochem ; 522: 53-60, 2017 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-28111303

RESUMO

The present work demonstrates the implementation of aptamers as capture molecules for a wide range of target classes in lateral flow assay applications. The targets were chosen in order to cover a wide range of target classes (small sized - metabolite, medium sized - protein, and large sized - whole cell/spore). For each target class one target molecule was selected as representative and appropriate aptamers were used for lateral flow assay development. The work points out that the implementation of aptamers as capture molecules in a universal lateral flow test platform was successful independent form target size. Furthermore, the limit of detection for p-aminohippuric acid in urine (200 ppm), lysozyme in white wine (20 ppm), and Alicyclobacillus spores in buffered orange juice (>8 CFU/mL) were determined using aptamers as capture molecules. The whole approach is considered as a proof of concept, regarding the ability of aptamers as an alternative to antibodies (in conjunction with directive 2010/63/EU on the protection of animals used for scientific purposes) in lateral flow applications.


Assuntos
Alicyclobacillus , Aptâmeros de Nucleotídeos/química , Sucos de Frutas e Vegetais/análise , Muramidase/análise , Esporos Bacterianos , Vinho/análise , Ácido p-Aminoipúrico/urina , Humanos
19.
Nephrology (Carlton) ; 22(8): 642-648, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27282888

RESUMO

AIM: Furosemide is a loop diuretic. Different authors demonstrated that continuous administration of furosemide modulates the expression of organic anion transporters. This study was undertaken to simultaneously evaluate the effects of furosemide pretreatment on organic anion transporter 1 (Oat1) and multidrug resistance protein 2 (Mrp2) renal expressions, on p-aminohippurate (PAH) pharmacokinetics and on renal and urinary PAH levels in rats. METHODS: Male Wistar rats were treated with furosemide (6 mg/100 g body weight per day, subcutaneously, 4 days) (treated group) or saline (control group). On the fifth day, PAH was administered as a bolus infusion in the femoral vein, and plasma samples were obtained from femoral artery at different time points. PAH levels in renal tissue and urine were also assessed. Renal Oat1 and Mrp2 expressions were evaluated by western blotting. RESULTS: Furosemide pretreatment increased both the expression of Oat1 and Mrp2. PAH plasma concentrations decreased following a biexponential function. The furosemide-treated group showed higher PAH plasma levels, a lower systemic clearance and elimination rate constant from the peripheral compartment, indicating that PAH renal elimination was decreased. PAH levels in renal tissue were significantly elevated and in urine appeared to be significantly lower as compared with control animals. CONCLUSIONS: Furosemide pretreatment caused a significant decrease of PAH renal elimination, despite Oat1 and Mrp2 augmented renal expression. The goal of the present study is the addition of important information in the wide gap of knowledge that exists about drug-drug interactions. Because of furosemide worldwide use, the data obtained are interesting and useful in terms of translation to clinical practice.


Assuntos
Furosemida/farmacologia , Rim/efeitos dos fármacos , Proteína 1 Transportadora de Ânions Orgânicos/efeitos dos fármacos , Inibidores de Simportadores de Cloreto de Sódio e Potássio/farmacologia , Ácido p-Aminoipúrico/farmacocinética , Transportadores de Cassetes de Ligação de ATP/efeitos dos fármacos , Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Interações Medicamentosas , Furosemida/administração & dosagem , Injeções Intravenosas , Injeções Subcutâneas , Rim/metabolismo , Masculino , Taxa de Depuração Metabólica , Modelos Biológicos , Proteína 1 Transportadora de Ânions Orgânicos/metabolismo , Ratos Wistar , Eliminação Renal/efeitos dos fármacos , Inibidores de Simportadores de Cloreto de Sódio e Potássio/administração & dosagem , Regulação para Cima , Ácido p-Aminoipúrico/administração & dosagem , Ácido p-Aminoipúrico/sangue , Ácido p-Aminoipúrico/urina
20.
Nutrients ; 9(1)2016 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-28036018

RESUMO

Supplementation of branched-chain amino acids (BCAA) has been demonstrated to promote skeletal muscle mass gain, but the mechanisms underlying this observation are still unknown. Since the regulation of muscle mass depends on a dynamic equilibrium (fasted losses-fed gains) in protein turnover, the aim of this study was to investigate the effects of BCAA supplementation on muscle protein synthesis and degradation in fed/fasted states and the related mechanisms. Fourteen 26- (Experiment 1) and 28-day-old (Experiment 2) piglets were fed reduced-protein diets without or with supplemental BCAA. After a four-week acclimation period, skeletal muscle mass and components of anabolic and catabolic signaling in muscle samples after overnight fasting were determined in Experiment 1. Pigs in Experiment 2 were implanted with carotid arterial, jugular venous, femoral arterial and venous catheters, and fed once hourly along with the intravenous infusion of NaH13CO3 for 2 h, followed by a 6-h infusion of [1-13C]leucine. Muscle leucine kinetics were measured using arteriovenous difference technique. The mass of most muscles was increased by BCAA supplementation. During feeding, BCAA supplementation increased leucine uptake, protein synthesis, protein degradation and net transamination. The greater increase in protein synthesis than in protein degradation resulted in elevated protein deposition. Protein synthesis was strongly and positively correlated with the intramuscular net production of α-ketoisocaproate (KIC) and protein degradation. Moreover, BCAA supplementation enhanced the fasted-state phosphorylation of protein translation initiation factors and inhibited the protein-degradation signaling of ubiquitin-proteasome and autophagy-lysosome systems. In conclusion, supplementation of BCAA to reduced-protein diet increases fed-state protein synthesis and inhibits fasted-state protein degradation, both of which could contribute to the elevation of skeletal muscle mass in piglets. The effect of BCAA supplementation on muscle protein synthesis is associated with the increase in protein degradation and KIC production in the fed state.


Assuntos
Aminoácidos de Cadeia Ramificada/administração & dosagem , Dieta com Restrição de Proteínas , Suplementos Nutricionais , Jejum , Proteínas Musculares/biossíntese , Músculo Esquelético/metabolismo , Aminoácidos de Cadeia Ramificada/sangue , Animais , Insulina/sangue , Cetoácidos/sangue , Leucina/sangue , Fosforilação , Biossíntese de Proteínas , Suínos , Ácido p-Aminoipúrico/sangue
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