RESUMEN
BACKGROUND/OBJECTIVES: Branched-chain amino acids (BCAAs) and purine nucleotide cycle (PNC) are both associated with energy metabolism. The purpose of this study was to explore the influences of BCAA supplementation on the PNC activity of male athletes in response to a bout of endurance running exercise. SUBJECTS/METHODS: Twelve male athletes (20.3±1.4 years) participated in the study. Each of the athletes received 12 g of a BCAA supplement (leucine 54%, isoleucine 19% and valine 27%) per day during the study. They performed two identical 60-min running exercises (65-70% maximum heart rate reserved) before and after receiving the BCAA supplements for 15 days. In addition to body composition measurement, plasma and urinary samples were also collected. Plasma samples were examined for the concentrations of glucose, lactate, BCAAs, alanine, glutamine, aspartate, hypoxanthine and uric acid. Urinary samples were examined for the concentrations of urea nitrogen, hydroxyproline, 3-methylhistidine and creatinine. RESULTS: Body composition and the concentrations of urinary metabolites were not affected by BCAA supplementation, whereas clearance of plasma lactate after recovery from exercise was enhanced by BCAA supplementation (P<0.05). Plasma aspartate concentration was increased (P<0.05), whereas plasma glutamine, hypoxanthine and uric acid concentrations were decreased (P<0.05) by BCAA supplementation. CONCLUSIONS: The findings suggest that BCAA supplements not only provided additional substrate to meet the energy demands of the athletes during endurance exercise but also reduced their PNC activity, and subsequently decreased uric acid production and reduced the incidence of gout in a person engaging in endurance exercise.
Asunto(s)
Aminoácidos de Cadena Ramificada/administración & dosificación , Nucleótidos de Purina/metabolismo , Carrera/fisiología , Fenómenos Fisiológicos en la Nutrición Deportiva , Adolescente , Aminoácidos de Cadena Ramificada/sangre , Antropometría , Atletas , Glucemia/metabolismo , Composición Corporal , Suplementos Dietéticos , Glutamina/sangre , Humanos , Hipoxantina/sangre , Isoleucina/administración & dosificación , Isoleucina/sangre , Leucina/administración & dosificación , Leucina/sangre , Masculino , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/metabolismo , Resistencia Física , Proyectos Piloto , Ácido Úrico/sangre , Urinálisis , Valina/administración & dosificación , Valina/sangre , Adulto JovenRESUMEN
To explore regulation of inosinic acid content in chicken meat as a result of feed additives, 576 one-day-old male Arbor Acres broilers were randomly allotted into 8 dietary treatments including control, purine nucleotide (P), betaine (B), soybean isoflavone (S), purine nucleotide + betaine (PB), purine nucleotide + soybean isoflavone (PS), betaine +soybean isoflavone (BS), and purine nucleotide + betaine + soybean isoflavone (PBS) by a 2 × 2 × 2 factorial arrangement. At d 42 of age, broilers were slaughtered, and growth performance, carcass characteristics, inosinic acid content, and activities of enzyme closely related to inosinic acid metabolism of broilers were measured. The results revealed that these feed additives did not affect ADG and ADFI of the broilers (P > 0.05). However, supplementing purine nucleotides lowered feed/gain of broilers in PS and PBS groups (P < 0.05). There was a significant interaction on feed/gain of broilers between purine nucleotides and soybean isoflavone (P < 0.05). The abdominal fat percentages in groups B, S, BS, and PBS were lower than the control group, respectively (P < 0.05). The thigh muscle percentages of groups P and B were higher than that of group PB (P < 0.05). There were certain interactions on the percentage of thigh muscle (P = 0.05) and abdominal fat (P < 0.05) between P, B, and S groups. Compared with the control group, inosinic acid content in broiler breast meat was improved by using feed additives (P < 0.05). Supplementing purine nucleotides, betaine, soybean isoflavone, and their combinations increased alkaline phosphatase activity in breast meat of broilers (P < 0.05). Purine nucleotides improved the activity of adenosine deaminase, but decreased the activity of 5'-nucleotidase. Soybean isoflavone lowered the activity of alkaline phosphatase. There were no significant interactions on activities of creatine kinase, adenosine deaminase, alkaline phosphatase, and 5'-nucleotidase between these additives (P > 0.05). The umami rating of broiler breast meat increased in conjunction with supplementing these additives. In conclusion, supplementing standard feed with the additives investigated in this study could improve inosinic acid content in chicken meat by increasing synthase activity or inhibiting degradation enzyme activity without inferior growth performance and carcass quality.
Asunto(s)
Betaína/metabolismo , Pollos/fisiología , Dieta/veterinaria , Inosina Monofosfato/metabolismo , Isoflavonas/metabolismo , Músculo Esquelético/química , Nucleótidos de Purina/metabolismo , Alimentación Animal/análisis , Animales , Betaína/administración & dosificación , Pollos/crecimiento & desarrollo , Suplementos Dietéticos/análisis , Isoflavonas/administración & dosificación , Masculino , Carne/análisis , Músculo Esquelético/enzimología , Músculo Esquelético/fisiología , Nucleótidos de Purina/administración & dosificación , Distribución Aleatoria , Glycine max/químicaRESUMEN
Using isolated potato tuber mitochondria possessing uncoupling protein (StUCP), we found that, under non-phosphorylating conditions, the sensitivity of aldehyde (all trans-retinal or 4-hydroxy-2-nonenal)-induced and fatty acid (linoleic acid)-induced StUCP-mediated proton leaks to GTP is controlled by the endogenous ubiquinone (Q) reduction level. The action of StUCP activators was abolished by GTP only when Q was sufficiently oxidized, but no inhibitory effect was observed when Q was highly reduced. Thus, the Q reduction level-dependent regulation of StUCP inhibition functions independently of the type of UCP activation and could be an important physiological factor affecting the efficiency of UCP-catalyzed uncoupling in plant mitochondria.
Asunto(s)
Aldehídos/farmacología , Ácido Linoleico/farmacología , Proteínas de Plantas/metabolismo , Ubiquinona/farmacología , Nucleótidos de Purina/metabolismo , Solanum tuberosum/efectos de los fármacos , Solanum tuberosum/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Kidney-yang deficiency syndrome (KYDS) is a diagnostic pattern in traditional Chinese medicine (TCM) and clinical data showed that the unbalance in adrenal cortical hormone is the key issue in KYDS patients. The processed Ranunculaceae aconitum carmichaeli debx (bai-fu-pian in Chinese, BFP) is one of the most commonly used Chinese herbs for treating KYDS. The present study was conducted to explore the therapeutic biomarkers of the BFP in treating hydrocortisone administration induced KYDS rats. MATERIALS AND METHODS: Thirty male Sprague-Dawley rats were randomly divided into five groups with six in each group. KYDS in rats was induced by i.p. injection of hydrocortisone at the dose of 10mg/kg per day for 15 days as described previously. The rats with KYDS were administered orally, starting from the day of hydrocortisone administration stopped, with BFP extract at the dose of 0.32g/kg, 0.64g/kg and 1.28g/kg per day respectively for 15 days. The blood samples were collected for the liquid chromatography quadruple time-of-flight mass spectrometry (LC-Q-TOF-MS) test, as well as radioimmunoassay to determine the concentrations of cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP) and adrenocorticotrophic hormone (ACTH). The metabolic responses to BFP administration were investigated by using the principal components analysis (PCA) and orthogonal partial least squares analysis (OPLS). Bioinformatics analyses were performed by using the Ingenuity Pathway Analysis (IPA). Variance analysis and linear regression analysis were used in this study. RESULTS: The signs and concentrations of cAMP, cGMP and ACTH in the model rats were similar to those previously described about KYDS rats and BFP treatment can reverse the changes. Seventeen significantly changed metabolites among different groups were identified. Thirteen metabolites were identified in the KYDS rats comparing to healthy rats with nine up-regulated and four down-regulated. After BFP treatment at three dosages, five up-regulated metabolites including phosphate, betaine, (4-hydroxyphenyl) acetaldehyde, 5-hydroxyindol-3-acetic acid and 5'-phosphoribosyl-N-formylglycinamide were dose-dependently reversed. The network analysis with IPA showed that four canonical pathways including superpathway of methionine degradation, purine nucleotides de novo biosynthesis II, tyrosine synthesis and serotonin receptor signaling involved the therapeutic mechanism of BFP in treating the KYDS rats. CONCLUSIONS: Five therapeutic biomarkers (phosphate, betaine, (4-hydroxyphenyl) acetaldehyde, 5-hydroxyindol-3-acetic acid and 5'-phosphoribosyl-N-formylglycinamide) and two corresponding canonical pathways (amino acid metabolism and purine nucleotide metabolism) were identified to be involved in the therapeutic mechanism of BFP treating the KYDS.
Asunto(s)
Aconitum/química , Enfermedades Renales/tratamiento farmacológico , Extractos Vegetales/farmacología , Deficiencia Yang/tratamiento farmacológico , Aminoácidos/metabolismo , Animales , Biomarcadores/metabolismo , Biología Computacional , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Hidrocortisona/toxicidad , Enfermedades Renales/fisiopatología , Modelos Lineales , Masculino , Espectrometría de Masas/métodos , Medicina Tradicional China , Extractos Vegetales/administración & dosificación , Análisis de Componente Principal , Nucleótidos de Purina/metabolismo , Ratas , Ratas Sprague-Dawley , Síndrome , Deficiencia Yang/fisiopatologíaRESUMEN
Vitamin C (ascorbic acid, AA) is a cofactor for many important enzymatic reactions and a powerful antioxidant. AA provides protection against oxidative stress by acting as a scavenger of reactive oxygen species, either directly or indirectly by recycling of the lipid-soluble antioxidant, α-tocopherol (vitamin E). Only a few species, including humans, guinea pigs, and zebrafish, cannot synthesize AA. Using an untargeted metabolomics approach, we examined the effects of α-tocopherol and AA deficiency on the metabolic profiles of adult zebrafish. We found that AA deficiency, compared with subsequent AA repletion, led to oxidative stress (using malondialdehyde production as an index) and to major increases in the metabolites of the purine nucleotide cycle (PNC): IMP, adenylosuccinate, and AMP. The PNC acts as a temporary purine nucleotide reservoir to keep AMP levels low during times of high ATP utilization or impaired oxidative phosphorylation. The PNC promotes ATP regeneration by converting excess AMP into IMP, thereby driving forward the myokinase reaction (2ADP â AMP + ATP). On the basis of this finding, we investigated the activity of AMP deaminase, the enzyme that irreversibly deaminates AMP to form IMP. We found a 47% increase in AMP deaminase activity in the AA-deficient zebrafish, complementary to the 44-fold increase in IMP concentration. These results suggest that vitamin C is crucial for the maintenance of cellular energy metabolism.
Asunto(s)
Antioxidantes/farmacología , Deficiencia de Ácido Ascórbico/metabolismo , Metabolismo Energético/efectos de los fármacos , Nucleótidos de Purina/metabolismo , Pez Cebra/metabolismo , alfa-Tocoferol/farmacología , Animales , Ácido Ascórbico/farmacología , Cobayas , HumanosRESUMEN
Besides regulating meristem formation and maintenance in vivo, SHOOTMERISTEMLESS (STM) has been shown to affect embryogenesis. While the over-expression of Brassica napus (Bn)STM enhances the number of microspore-derived embryos produced in culture and their ability to regenerate viable plants, a down-regulation of this gene represses the embryogenic process (Elhiti et al., J Exp Bot, 61:4069-4085, 2010). Synthesis and degradation of pyrimidine and purine nucleotides were measured in developing microspore-derived embryos (MDEs) generated from B. napus lines ectopically expressing or down-regulating BnSTM. Pyrimidine metabolism was investigated by following the metabolic fate of exogenously supplied (14)C-uridine, uracil and orotic acid, whereas purine metabolism was estimated by using (14)C-adenine, adenosine and inosine. The improvement in embryo number and quality affected by the ectopic expression of BnSTM was linked to the increased pyrimidine and purine salvage activity during the early phases of embryogenesis and the enlargement of the adenylate pool (ATP + ADP) required for the active growth of the embryos. This was due to an increase in transcriptional and enzymatic activity of several salvage enzymes, including adenine phosphoribosyltransferase (APRT) and adenosine kinase (ADK). The highly operative salvage pathway induced by the ectopic expression of BnSTM was associated with a slow catabolism of nucleotides, suggesting the presence of an antagonist mechanism controlling the rate of salvage and degradation pathways. During the second half of embryogenesis utilization of uridine for UTP + UDPglucose (UDPG) synthesis increased in the embryos over-expressing BnSTM, and this coincided with a better post-germination performance. All these events were precluded by the down-regulation of BnSTM which repressed the formation of the embryos and their post-embryonic performance. Overall, this work provides evidence that precise metabolic changes are associated with proper embryo development in culture.
Asunto(s)
Brassica napus/embriología , Brassica napus/fisiología , Proteínas de Plantas/genética , Nucleótidos de Purina/metabolismo , Nucleósidos de Pirimidina/metabolismo , Adenina Fosforribosiltransferasa/metabolismo , Adenosina Quinasa/metabolismo , Transporte Biológico , Brassica napus/citología , Brassica napus/genética , Isótopos de Carbono/análisis , Regulación hacia Abajo , Expresión Génica/genética , Germinación , Meristema/citología , Meristema/embriología , Meristema/genética , Meristema/fisiología , Ácido Orótico/metabolismo , Proteínas de Plantas/metabolismo , Polen/fisiología , Factores de Tiempo , Uracilo/metabolismo , Uridina/metabolismoRESUMEN
To determine the metabolic profiles of purine nucleotides and related compounds in leaves and roots of tea (Camellia sinensis), we studied the in situ metabolic fate of 10 different (14)C-labeled precursors in segments from tea seedlings. The activities of key enzymes in tea leaf extracts were also investigated. The rates of uptake of purine precursors were greater in leaf segments than in root segments. Adenine and adenosine were taken up more rapidly than other purine bases and nucleosides. Xanthosine was slowest. Some adenosine, guanosine and inosine was converted to nucleotides by adenosine kinase and inosine/guanosine kinase, but these compounds were easily hydrolyzed, and adenine, guanine and hypoxanthine were generated. These purine bases were salvaged by adenine phosphoribosyltransferase and hypoxanthine/guanine phosphoribosyltransferase. Salvage activity of adenine and adenosine was high, and they were converted exclusively to nucleotides. Inosine and hypoxanthine were salvaged to a lesser extent. In situ (14)C-tracer experiments revealed that xanthosine and xanthine were not salvaged, although xanthine phosphoribosyltransferase activity was found in tea extracts. Only some deoxyadenosine and deoxyguanosine was salvaged and utilized for DNA synthesis. However, most of these deoxynucleosides were hydrolyzed to adenine and guanine and then utilized for RNA synthesis. Purine alkaloid biosynthesis in leaves is much greater than in roots. In situ experiments indicate that adenosine, adenine, guanosine, guanine and inosine are better precursors than xanthosine, which is a direct precursor of a major pathway of caffeine biosynthesis. Based on these results, possible routes of purine metabolism are discussed.
Asunto(s)
Camellia sinensis/metabolismo , Nucleótidos de Purina/metabolismo , Plantones/metabolismo , Camellia sinensis/enzimología , Redes y Vías Metabólicas , Hojas de la Planta/metabolismo , Raíces de Plantas/metabolismo , Nucleótidos de Purina/biosíntesis , Plantones/enzimologíaRESUMEN
To find general metabolic profiles of purine ribo- and deoxyribonucleotides in potato (Solanum tuberosum L.) plants, we looked at the in situ metabolic fate of various (14)C-labelled precursors in disks from growing potato tubers. The activities of key enzymes in potato tuber extracts were also studied. Of the precursors for the intermediates in de novo purine biosynthesis, [(14)C]formate, [2-(14)C]glycine and [2-(14)C]5-aminoimidazole-4-carboxyamide ribonucleoside were metabolised to purine nucleotides and were incorporated into nucleic acids. The rates of uptake of purine ribo- and deoxyribonucleosides by the disks were in the following order: deoxyadenosine > adenosine > adenine > guanine > guanosine > deoxyguanosine > inosine > hypoxanthine > xanthine > xanthosine. The purine ribonucleosides, adenosine and guanosine, were salvaged exclusively to nucleotides, by adenosine kinase (EC 2.7.1.20) and inosine/guanosine kinase (EC 2.7.1.73) and non-specific nucleoside phosphotransferase (EC 2.7.1.77). Inosine was also salvaged by inosine/guanosine kinase, but to a lesser extent. In contrast, no xanthosine was salvaged. Deoxyadenosine and deoxyguanosine, was efficiently salvaged by deoxyadenosine kinase (EC 2.7.1.76) and deoxyguanosine kinase (EC 2.7.1.113) and/or non-specific nucleoside phosphotransferase (EC 2.7.1.77). Of the purine bases, adenine, guanine and hypoxanthine but not xanthine were salvaged for nucleotide synthesis. Since purine nucleoside phosphorylase (EC 2.4.2.1) activity was not detected, adenine phosphoribosyltransferase (EC 2.4.2.7) and hypoxanthine/guanine phosphoribosyltransferase (EC 2.4.2.8) seem to play the major role in salvage of adenine, guanine and hypoxanthine. Xanthine was catabolised by the oxidative purine degradation pathway via allantoin. Activity of the purine-metabolising enzymes observed in other organisms, such as purine nucleoside phosphorylase (EC 2.4.2.1), xanthine phosphoribosyltransferase (EC 2.4.2.22), adenine deaminase (EC 3.5.4.2), adenosine deaminase (EC 3.5.4.4) and guanine deaminase (EC 3.5.4.3), were not detected in potato tuber extracts. These results suggest that the major catabolic pathways of adenine and guanine nucleotides are AMP --> IMP --> inosine --> hypoxanthine --> xanthine and GMP --> guanosine --> xanthosine --> xanthine pathways, respectively. Catabolites before xanthosine and xanthine can be utilised in salvage pathways for nucleotide biosynthesis.
Asunto(s)
Tubérculos de la Planta/metabolismo , Purinas/metabolismo , Solanum tuberosum/metabolismo , Adenina/metabolismo , Desoxirribonucleósidos/metabolismo , Guanina/metabolismo , Guanosina/metabolismo , Hipoxantina Fosforribosiltransferasa/metabolismo , Inosina/metabolismo , Redes y Vías Metabólicas , Modelos Biológicos , Pentosiltransferasa/metabolismo , Nucleótidos de Purina/metabolismo , Purina-Nucleósido Fosforilasa/metabolismo , Purinas/biosíntesis , Ribonucleósidos/metabolismo , Xantina/metabolismo , XantinasRESUMEN
Hyperbaric oxygen treatment has been suggested as able to reduce hypoxia induced neuronal damage. The aim of the study was to compare the impact of different reoxygenation strategies on early metabolical (purine nucleotide content determined by HPLC) and morphological changes (index of cell injury after celestine blue/acid fuchsin staining) of hypoxically damaged rat neocortical brain slices. For this purpose slices (300 microm and 900 microm) were subjected to either 5 or 30 min of hypoxia by gassing the incubation medium with nitrogen. During the following reoxygenation period treatment groups were administered either 100% oxygen (O) or room air (A) at normobaric (1 atm absolute, NB-O; NB-A) or hyperbaric (2.5 atm absolute, HB-O; HB-A) conditions. After 5 min of hypoxia, both HB-O and NB-O led to a complete nucleotide status restoration (ATP/ADP; GTP/GDP) in 300 microm slices. However, reoxygenation after 30 min of hypoxia was less effective, irrespective of the oxygen pressure. Furthermore, administering hyperbaric room air resulted in no significant posthypoxic nucleotide recovery. In 900 microm slices, both control incubation as well as 30 min of hypoxia resulted in significantly lower trinucleotide and higher dinucleotide levels compared to 300 microm slices. While there was no significant difference between HB-O and NB-O on the nucleotide status, morphological evaluation revealed a better recovery of the index of cell injury (profoundly injured/intact cell-ratio) in the HB-O group. Conclusively, the posthypoxic recovery of metabolical characteristics was dependent on the duration of hypoxia and slice thickness, but not on the reoxygenation pressure. A clear restorative effect on purine nucleotides was found only in early-administered HB-O as well as NB-O in contrast to room air treated slices. However, these pressure independent metabolic changes were morphologically accompanied by a significantly improved index of cell injury, indicating a possible neuroprotective role of HB-O in early posthypoxic reoxygenation.
Asunto(s)
Química Encefálica/fisiología , Encéfalo/patología , Oxigenoterapia Hiperbárica , Hipoxia-Isquemia Encefálica/metabolismo , Hipoxia-Isquemia Encefálica/patología , Terapia por Inhalación de Oxígeno , Nucleótidos de Purina/metabolismo , Adenosina Difosfato/metabolismo , Adenosina Trifosfato/metabolismo , Animales , Recuento de Células , Supervivencia Celular/fisiología , Cromatografía Líquida de Alta Presión , Colorantes , Metabolismo Energético/fisiología , Guanosina Difosfato/metabolismo , Guanosina Trifosfato/metabolismo , Técnicas In Vitro , Masculino , Neocórtex/metabolismo , Neocórtex/patología , Ratas , Ratas WistarRESUMEN
Protective strategies to minimize the hematological toxicity in connection with bone marrow transplantation (BMT) have been successful, but toxicity to the gastrointestinal tract prevents further dose escalation and therefore limits the application of the treatment. As it is known that chemotherapy leads to disruption of the intestinal barrier and morphological changes of mitochondria in enterocytes, this study was conducted in order to investigate intestinal energy metabolism and permeability after intensive cytotoxic therapy in rats. Intestinal damage was produced by intraperitoneal administration of the cytostatic etoposide. Intestinal permeability was assessed by a [51Cr]EDTA absorption test and intestinal purine nucleotide content by a high-performance liquid chromatography (HPLC) technique. Four hours after the administration of etoposide, and the next 48 hr, there was a significant increase in the intestinal permeability (P < 0.05) and a significant reduction of the purine nucleotide content in the intestinal epithelial cells (P < 0.01) as compared to control animals. This early disturbance in enterocyte energy metabolism may be a key event in the development of the intestinal damage, induced by chemotherapy, and an explanation for the early disruption of the intestinal barrier demonstrable before morphological changes are evident.
Asunto(s)
Antineoplásicos/efectos adversos , Modelos Animales de Enfermedad , Etopósido/efectos adversos , Enfermedades Intestinales/inducido químicamente , Enfermedades Intestinales/metabolismo , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/metabolismo , Nucleótidos de Purina/análisis , Nucleótidos de Purina/metabolismo , Animales , Antineoplásicos/administración & dosificación , Permeabilidad Capilar/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Evaluación Preclínica de Medicamentos , Metabolismo Energético/efectos de los fármacos , Enterocitos/efectos de los fármacos , Enterocitos/metabolismo , Etopósido/administración & dosificación , Inyecciones Intraperitoneales , Mucosa Intestinal/química , Recuento de Leucocitos , Masculino , Ratas , Ratas Endogámicas Lew , Factores de TiempoRESUMEN
In this study we have tested the hypothesis that precursors of the purine base backbone--glutamine, glycine, aspartic acid, and formate--promote meiotic arrest when included in medium containing established meiotic inhibitors and that this occurs in glucose-dependent fashion. An initial experiment established that in medium supplemented with 4 mM hypoxanthine and containing no purine precursors, very little meiotic arrest was maintained in cumulus cell-enclosed oocytes after 17-18 hr (90% germinal vesicle breakdown; GVB). Increasing concentrations of glucose reduced the maturation percentage such that only 57% had matured at 0.55 mM. The addition of 2 mM glutamine (Gln) alone reduced the maturation percentage in the absence of glucose (70% GVB), and the further addition of glucose revealed an additive inhibitory effect between these two supplements. Dose response experiments with Gln, glycine (Gly), aspartic acid and formate showed that in medium supplemented with hypoxanthine, very little inhibitory action was observed in the absence of glucose but that upon addition of this hexose, a dramatic decrease in maturation percentage was observed in the Gln and Gly groups. Results of experiments using combinations of precursors showed that when Gln and Gly were added together, greater augmentation of meiotic arrest maintained by either hypoxanthine or dibutyryl cAMP was achieved in the presence of glucose than with either amino acid alone. The addition of purine precursors significantly increased the extent of purine nucleotide production by oocyte-cumulus cell complexes, and this was accentuated by glucose. It is concluded that the presence of purine precursors can augment the meiosis-arresting action of established meiotic inhibitors in glucose-dependent fashion, and that this is due, at least in part, to their incorporation into purine nucleotides via the de novo synthetic pathway.
Asunto(s)
Bucladesina/farmacología , Hipoxantina/farmacología , Oocitos/efectos de los fármacos , Purinas/química , Animales , Ácido Aspártico/farmacología , Bucladesina/química , Relación Dosis-Respuesta a Droga , Formiatos/farmacología , Glucosa/administración & dosificación , Glucosa/farmacología , Glutamina/farmacología , Glicina/farmacología , Hipoxantina/química , Meiosis/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Modelos Químicos , Oocitos/crecimiento & desarrollo , Oocitos/metabolismo , Nucleótidos de Purina/metabolismo , Relación Estructura-ActividadRESUMEN
The effect of intravenously administered ascorbate on the ischemic and reperfused rat skeletal muscle was investigated. Purine nucleotides and phospholipids in skeletal muscle from rats subjected to 4 h of ischemia followed by 1-h reperfusion were analyzed by high-performance liquid chromatography. In addition, ATP, phosphocreatine (PCr), Pi, and phosphomonoesters (PME) were analyzed by 31P-nuclear magnetic resonance at 202.4 MHz, and individual PME such as glucose-6-phosphate and IMP were quantified. PCr and ATP were exhausted after 4 h of ischemia and recovered poorly upon reperfusion in the soleus and tibialis muscle of untreated rats. Postischemic reperfusion resulted in significant loss of cardiolipin. Treatment with 55 mM ascorbate resulted in total restoration of PCr during reperfusion, and ATP recovered to 42% of control in the soleus. Recovery was improved in the tibialis as well, and the cardiolipin decrease was limited. A lower ascorbate concentration (5 mM) did not enhance postischemic recovery. Our findings show that a high dose of ascorbate improves the energetic state of rat skeletal muscle during postischemic reperfusion, probably due to its antioxidant function.
Asunto(s)
Ácido Ascórbico/farmacología , Isquemia/metabolismo , Músculo Esquelético/irrigación sanguínea , Fosfolípidos/metabolismo , Nucleótidos de Purina/metabolismo , Reperfusión , Animales , Cardiolipinas/metabolismo , Bovinos , Cromatografía Líquida de Alta Presión , Metabolismo Energético , Femenino , Espectroscopía de Resonancia Magnética , Músculo Esquelético/efectos de los fármacos , Fosfatos/metabolismo , Fosfatidiletanolaminas/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Activity of NADP-dependent isocitrate- and malate dehydrogenases and the rate of NADP reduction into NAD+ were studied in various rat brain compartments during the hypoglycemic coma and within various periods after the coma cupping using glucose administration. Severe hypoglycemia was accompanied by distinct decrease in content of NADH x H+ in nervous tissue, while the enzymatic activity studied was not altered and content of NADPH x H+ was not decreased. Within the early restoration period activity of cytoplasmic NADP-malate dehydrogenase and content of NADPH x H+ were increased in brain tissue. Production of NADPH x H+ during hypoglycemic coma and after its cupping was maintained in nervous tissue at the adequately high level and did not affect the restoration-related synthesis.
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Encéfalo/enzimología , Hipoglucemia/enzimología , Insulina/efectos adversos , NADPH Deshidrogenasa/metabolismo , Nucleótidos de Purina/metabolismo , Animales , Encéfalo/metabolismo , Hipoglucemia/etiología , Isocitrato Deshidrogenasa/metabolismo , Malato Deshidrogenasa/metabolismo , Masculino , NADP/metabolismo , Oxidación-Reducción , RatasRESUMEN
The gastric mucosal microcirculation and purine nucleotide metabolism were studied in rats after hemorrhagic shock and retransfusion. The mucosal surface density of perfused vessels (SDPV) and the mucosal levels of ATP, ADP, AMP, IMP, hypoxanthine and uric acid were measured following 15 min of hemorrhagic shock and 10 and 30 min after retransfusion, and the effects of pretreatment with allopurinol or ascorbate were studied. During shock there was a dephosphorylation of nucleotides and a decline in the SDPV. Retransfusion led to an additional reduction in the SDPV, but a complete restoration of preshock nucleotide levels 30 min after retransfusion. Allopurinol accelerated early rephosphorylation of nucleotides without effects upon SDPV while ascorbate completely preserved the mucosal level of energy-rich nucleotides 15 min after hemorrhagic shock and increased SDPV during early reperfusion. The results showed that there was a renewal of energy stores in gastric mucosa after hemorrhagic shock and retransfusion although parts of the vascular bed were not reperfused. The mucosal energy depletion after 15 min of hemorrhagic shock and part of the mucosal vessel injury after retransfusion were prevented by pretreatment with ascorbate.
Asunto(s)
Alopurinol/uso terapéutico , Ácido Ascórbico/uso terapéutico , Mucosa Gástrica/irrigación sanguínea , Mucosa Gástrica/metabolismo , Daño por Reperfusión/prevención & control , Choque Hemorrágico/tratamiento farmacológico , Animales , Transfusión de Sangre Autóloga , Masculino , Microcirculación , Nucleótidos de Purina/análisis , Nucleótidos de Purina/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Gastric mucosal microcirculation and purine nucleotide metabolism were studied after hemorrhagic shock and retransfusion in rats. The number of perfused microvessels and the concentration of adenosine triphosphate, adenosine monophosphate, inosine monophosphate, inosine, hypoxanthine, xanthine, and uric acid were investigated in mucosal biopsy specimens after 15 or 45 min of hemorrhagic shock and after 15 min of shock followed by 30 min of retransfusion. During shock a dephosphorylation of nucleotides and a decrease in the number of perfused microvessels occurred, the more the longer the duration of the shock period. Retransfusion led to an additional reduction in the number of perfused microvessels, but there was a partial restoration of high-energy phosphate metabolites in those areas of the mucosa which maintained a blood flow. The results indicate that there is a renewal of energy stores in gastric mucosa after hemorrhagic shock and reperfusion, although parts of the vascular bed are not reperfused.
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Mucosa Gástrica/irrigación sanguínea , Mucosa Gástrica/metabolismo , Nucleótidos de Purina/metabolismo , Reperfusión , Choque Hemorrágico/metabolismo , Choque Hemorrágico/fisiopatología , Animales , Transfusión de Sangre Autóloga , Mucosa Gástrica/patología , Masculino , Microcirculación , Ratas , Ratas Sprague-Dawley , Choque Hemorrágico/terapiaRESUMEN
The effect of carbohydrate (CHO) ingestion on metabolic responses to exercise has been investigated. Subjects cycled at approximately 70% of maximal oxygen uptake to fatigue [135 +/- 17 (+/- SE) min] on the first occasion (control, CON) and at the same work load and duration on the second occasion but with addition of ingestion of CHO during the exercise. Biopsies were taken from the quadriceps femoris muscle before and after exercise. The sum of the hexose monophosphates (HMP), as well as lactate and alanine, in muscle was higher after CHO exercise (P less than or equal to 0.05, P less than or equal to 0.05, and P less than or equal to 0.01, respectively). Acetylcarnitine increased during exercise but was not significantly different between treatments after exercise (CON, 6.6 +/- 1.7; CHO, 10.0 +/- 1.2 mmol/kg dry wt; P = NS). The sum of the tricarboxylic acid cycle intermediates (TCAI; citrate + malate + fumarate) was increased during exercise and was higher after CHO exercise (2.34 +/- 0.32 vs. 1.68 +/- 0.17 mmol/kg dry wt; P less than or equal to 0.05). IMP was less than 0.1 mmol/kg dry wt at rest and increased to 0.77 +/- 0.26 (CON) and 0.29 +/- 0.11 mmol/kg dry wt (CHO) (P less than or equal to 0.05) during exercise. It was recently found that during prolonged exercise there is initially a rapid and large expansion of TCAI and glycogenolytic intermediates in human muscle followed by a continuous decline in TCAI and glycogenolytic intermediates [K. Sahlin, A. Katz, and S. Broberg. Am. J. Physiol. 259 (Cell Physiol. 28): C834-C841, 1990].(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Carbohidratos de la Dieta/administración & dosificación , Inosina Monofosfato/metabolismo , Músculos/metabolismo , Esfuerzo Físico/fisiología , Adulto , Glucemia/metabolismo , Carnitina/metabolismo , Carbohidratos de la Dieta/metabolismo , Ácidos Grasos/sangre , Femenino , Glucógeno/metabolismo , Frecuencia Cardíaca , Humanos , Insulina/sangre , Masculino , Persona de Mediana Edad , Contracción Muscular , NAD/metabolismo , Oxidación-Reducción , Fosfatos/metabolismo , Nucleótidos de Purina/metabolismo , Respiración , Ácidos Tricarboxílicos/metabolismoRESUMEN
Although several different enzymes with 5'-nucleotidase activity have been described in mammalian cells, their functions in nucleotide metabolism have not been clearly distinguished. In the present experiments, a mutant human T lymphoblastoid cell line (CEM-dAdoR) was selected specifically for resistance to deoxyadenosine toxicity. Compared to parental CEM cells, the variant had 4-fold elevated ATP-activated cytosolic 5'-nucleotidase activity. Other enzymes of potential importance for deoxyadenosine metabolism were indistinguishable in the two cell types. In medium supplemented with the adenosine deaminase inhibitor deoxycoformycin, the T cells with increased 5'-nucleotidase accumulated less nucleotides from exogenously added deoxyadenosine, or 9-beta-D-arabinofuranosyladenine, than did parental T lymphocytes. These metabolic changes were associated with resistance to the growth inhibitory effects of these nucleosides, and also to deoxyguanosine and to 9-beta-D-arabinofuranosylguanine. The T cells with elevated 5'-nucleotidase activity formed more 2',3'-dideoxyadenosine than did parental cells, in deoxycoformycin-supplemented medium. The accumulation of 2',3'-dideoxyadenosine 5'-triphosphate from 2',3'-dideoxyinosine was similarly augmented in the mutant. These data establish the importance of the cytosolic 5'-nucleotidase for the metabolism of purine 2'-deoxyribonucleosides, arabinonucleosides and 2',3'-dideoxyribonucleosides in T lymphoblasts.
Asunto(s)
5'-Nucleotidasa/análisis , Desoxiadenosinas/farmacología , Linfocitos T/efectos de los fármacos , Linfocitos T/enzimología , Adenosina Trifosfato/fisiología , Antivirales/metabolismo , Arabinonucleósidos/metabolismo , Línea Celular , Desoxiadenosinas/toxicidad , Didesoxinucleósidos/metabolismo , Didesoxinucleósidos/farmacología , Resistencia a Medicamentos , Activación Enzimática , VIH/efectos de los fármacos , Humanos , Nucleósidos de Purina/metabolismo , Nucleósidos de Purina/farmacología , Nucleótidos de Purina/metabolismoRESUMEN
Patients with McArdle's disease (myophosphorylase deficiency) cannot use muscle glycogen as an energy source during exercise. They therefore are an ideal model to learn about the metabolic adaptations which develop during endurance exercise leading to glycogen depletion. This review summarizes the current knowledge of ammonia and amino acid metabolism in these patients and also adds several new data. During incremental exercise tests in patients with McArdle's disease, forearm venous plasma ammonia concentration rises to a value between 200 and 500 microM. Femoral arteriovenous difference studies show that muscle produces the ammonia. The leg release of both ammonia and glutamine (in mumol/min) has been estimated to be five- to tenfold larger in one of these patients than in healthy individuals exercising at comparable relative work load. Patients with McArdle's disease have a larger uptake of branched-chain amino acids (BCAA) by exercising leg muscles and show a more rapid activation of the muscle branched-chain 2-oxo acid dehydrogenase complex, a key enzyme in the degradation of the BCAA. In general, supplements of BCAA taken before the exercise test lead to a deterioration of exercise performance and a higher increase in heart rate and plasma ammonia during exercise, whereas supplements of branched-chain 2-oxo acids improve exercise performance and lead to a smaller increase in heart rate and plasma ammonia. At constant power output, patients with McArdle's disease show a rapid increase in heart rate and exertion perceived in the exercising muscles, which peak within 10 min after the start of exercise and then fall again ("second wind"). Peak heart rate and peak exertion coincide with a peak in plasma ammonia. Ammonia production during exercise in these patients is estimated to exceed the reported breakdown of ATP to IMP and therefore most likely originates from the metabolism of amino acids. Deamination of amino acids via the reactions of the purine nucleotide cycle and glutamate dehydrogenase are possible pathways. Deamination of glutamine, released by muscle, by glutaminase present in the endothelial cells of the vascular system may also contribute to the ammonia production. The observations made in these patients have led to the hypothesis that excessive acceleration of the metabolism of BCAA drains 2-oxoglutarate in the primary aminotransferase reaction and thus reduces flux in the citric acid cycle and impedes aerobic oxidation of glucose and fatty acids. This draining effect is normally counteracted by the anaplerotic conversion of muscle glycogen to citric acid cycle intermediates, a reaction which is severely hampered in these patients due to the glycogen breakdown defect.(ABSTRACT TRUNCATED AT 400 WORDS)
Asunto(s)
Aminoácidos de Cadena Ramificada/metabolismo , Amoníaco/metabolismo , Ejercicio Físico/fisiología , Enfermedad del Almacenamiento de Glucógeno Tipo V/metabolismo , Aminoácidos de Cadena Ramificada/sangre , Amoníaco/sangre , Animales , Desaminación , Enfermedad del Almacenamiento de Glucógeno Tipo V/sangre , Humanos , Masculino , Contracción Muscular , Músculos/enzimología , Músculos/metabolismo , Resistencia Física/fisiología , Nucleótidos de Purina/metabolismo , RatasRESUMEN
The purine precursor AICAR (5-amino-4-imidazolecarboxamide) has been advocated as a substrate for myocardial adenine nucleotide repletion during postischemic reperfusion. The purpose of this study was to investigate the acute effects of this agent on adenine nucleotides, inosine monophosphate, and postischemic ventricular function in an isolated rat heart preparation. The hearts were perfused at constant flow, either continuously for 90 minutes or for a 30 minute period followed by 10 minutes of global normothermic (37 degrees C) ischemia. The ischemic hearts were then reperfused for 15, 30, and 60 minutes. Both groups were treated with AICAR in a concentration of 100 mumol/L throughout the perfusion protocols. In the nonischemic time control group there was no effect on the levels of adenosine nucleotides or developed pressure over 90 minutes of perfusion. In contrast, AICAR treatment increased tissue inosine monophosphate content four-fold and sevenfold at 60 and 90 minutes, respectively (p less than 0.05), but had no effect on tissue adenosine monophosphate levels. During ischemia, there was a 50% decrease in adenosine triphosphate content in the AICAR-treated hearts and a thirteen-fold increase in adenosine monophosphate levels (p less than 0.05). After 60 minutes of reperfusion, adenosine triphosphate and monophosphate levels in the AICAR-treated hearts recovered to only 52% and 59% of preischemic values, respectively. These findings were similar to those observed in the untreated ischemic hearts. In contrast, tissue inosine monophosphate content in the AICAR-treated hearts during reperfusion remained significantly elevated and was fivefold greater than the reperfusion values in the untreated group. Concurrently, AICAR failed to enhance the recovery of postischemic left ventricular developed pressure. These results suggest that inhibition of the conversion of inosine monophosphate to adenosine monophosphate limits the usefulness of the agent in evaluating the temporal relationships between postischemic adenosine triphosphate repletion and recovery of myocardial function in the acute setting.