RESUMEN
AIM: Bronchopulmonary dysplasia (BPD), a respiratory complication associated with neonatal prematurity, presents opportunities for pharmacological intervention due to its contributing risk factors. Despite diuretics' controversial usage in BPD treatment and varying institutional practices, this review aims to consolidate evidence from clinical trials regarding diuretic use in BPD. METHODS: We conducted a systematic review following PRISMA guidelines, searching EMBASE, Medline, Web of Science and CINAHL databases (PROSPERO 2022: CRD42022328292). Covidence facilitated screening and data extraction, followed by analysis and formatting in Microsoft Excel. RESULTS: Among 430 screened records, 13 were included for analysis. Three studies assessed spironolactone and chlorothiazide combinations, two studied spironolactone and hydrochlorothiazide, while eight examined furosemide. All studies evaluated drug effects on dynamic pulmonary compliance and pulmonary resistance, serving as comparative measures in our review. CONCLUSION: Diuretics' effectiveness in treating bronchopulmonary dysplasia remains uncertain. The limited number of identified randomised controlled trials (RCTs) hampers high-level evidence-based conclusions when applying the Population, Intervention, Comparison, Outcome (PICO) approach. Conducting large prospective studies of good quality could provide more definitive insights, but the rarity of outcomes and eligible patients poses challenges. Further research, primarily focusing on RCTs assessing diuretics' safety and efficacy in this population, is warranted.
Asunto(s)
Displasia Broncopulmonar , Diuréticos , Recien Nacido Prematuro , Displasia Broncopulmonar/tratamiento farmacológico , Humanos , Recién Nacido , Diuréticos/uso terapéuticoRESUMEN
The Neuropathology of Human Parechovirus (HPeV) is not widely described due to the relatively recent discovery of the virus combined with a limited number of autopsy case reports. We report the case of an infant boy born at 38 weeks who, six days after birth, presented with fever and severe neurological dysfunction. Human Parechovirus Type 3 (HPeV3) RNA was detected in his cerebrospinal fluid (CSF) and blood. He died five days after his initial presentation. Neuropathologic examination demonstrated multicystic encephalomalacia (ME). This case report confirms that white matter pathology is dominant in HPeV3 infection. A unique feature, of HPeV encephalomalacia is absence of CSF pleocytosis and minimal inflammation in the meninges. The findings permit comment on the pathogenesis of brain injury by this virus.
Asunto(s)
Encefalomalacia/patología , Encefalomalacia/virología , Parechovirus , Infecciones por Picornaviridae/patología , Encefalomalacia/diagnóstico , Resultado Fatal , Humanos , Recién Nacido , Masculino , Parechovirus/aislamiento & purificación , Infecciones por Picornaviridae/diagnósticoRESUMEN
Our aim was to assess the utility of serum thyroxine and thyroid stimulating hormone performed at 10-14 days of life in diagnosing congenital hypothyroidism (CH) in babies born to mothers with hypothyroidism. This was a retrospective study of all babies born in a tertiary referral centre for neonatology over a 12-month period. Infants who had thyroid function testing (TFT) checked at 10-14 days of life because of maternal hypothyroidism during the period of study were included. The results of the newborn bloodspot and day 10-14 TFT were recorded along with whether or not patients were subsequently treated. Of the 319 patients included in the study, only two patients were found to have CH and in both cases the newborn blood spot had been abnormal. CONCLUSION: No extra cases of CH were detected from the thyroid test at 10-14 days and this practice should be discontinued due to the robust nature of existing newborn screening programmes. What is Known: ⢠Congenital hypothyroidism(CH) is the commonest preventable cause of childhood intellectual impairment. ⢠Family history of hypothyroidism has been implicated as a risk factor for CH. ⢠CH has formed part of newborn screening since the 1970s. What is New: ⢠There is no research recommending thyroid function testing at 10-14 days of life to detect CH in neonates born to mothers with hypothyroidism. ⢠Thyroid function testing at 10-14 days of life does not improve diagnostic yield for CH in babies born to mothers with hypothyroidism. ⢠Newborn blood spot remains the mainstay for accurate and timely diagnosis of CH.
Asunto(s)
Hipotiroidismo Congénito/diagnóstico , Hipotiroidismo , Complicaciones del Embarazo , Pruebas de Función de la Tiroides , Hipotiroidismo Congénito/sangre , Femenino , Humanos , Recién Nacido , Tamizaje Neonatal , Embarazo , Estudios Retrospectivos , Factores de RiesgoRESUMEN
Helicobacter pylori is a risk factor for the development of gastritis, gastroduodenal ulcers, and gastric adenocarcinoma. H. pylori-induced disruption of epithelial adherens junctions (AJs) is thought to promote the development of severe disease; however, the mechanisms whereby H. pylori alters AJ structure remain incompletely understood. The present study demonstrates that H. pylori infection in human patients is associated with elevated serum levels of an 80-kDa E-cadherin ectodomain, whose presence is independent of the presence of serum antibodies against CagA. In vitro, a heat-labile H. pylori surface component activates the host protease calpain in human gastric MKN45 cells independently of the virulence factors CagA and VacA. H. pylori-induced calpain activation results in cleavage of E-cadherin to produce a 100-kDa truncated form and induce relocalization of E-cadherin and ß-catenin. Stimulation of MKN45 cells with the toll-like receptor 2 (TLR2) ligand P3C activated calpain and disrupted E-cadherin and ß-catenin in a pattern similar to that induced by H. pylori. Inhibition of TLR2 prevented H. pylori-induced calpain activation and AJ disassembly. Together, these findings identify a novel pathway whereby H. pylori activates calpain via TLR2 to disrupt gastric epithelial AJ structure.
Asunto(s)
Uniones Adherentes/patología , Calpaína/metabolismo , Mucosa Gástrica/microbiología , Helicobacter pylori/patogenicidad , Interacciones Huésped-Patógeno , Receptor Toll-Like 2/metabolismo , Cadherinas/sangre , Cadherinas/metabolismo , Línea Celular , Línea Celular Tumoral , Activación Enzimática , Células Epiteliales/microbiología , Células Epiteliales/patología , Mucosa Gástrica/citología , Mucosa Gástrica/patología , Infecciones por Helicobacter/sangre , Infecciones por Helicobacter/microbiología , Helicobacter pylori/metabolismo , Humanos , beta Catenina/sangre , beta Catenina/metabolismoRESUMEN
Helicobacter pylori infects more than half of the human population worldwide. In the absence of treatment, this persistent infection leads to asymptomatic gastritis, which in some cases can progress into gastric ulcers and adenocarcinomas. The host-microbial interactions that govern the clinical outcome of infection remain incompletely understood. H. pylori is known to disrupt gastric epithelial tight junctions, which may represent a significant component of disease pathogenesis. The present study demonstrates that H. pylori disrupt epithelial tight junctional claudin-4 in a Rho kinase (ROCK)-dependent manner in human gastric epithelial (HGE-20) cell monolayers, independently of the virulence factors CagA and VacA, and without altering claudin-4 transcription. In the same epithelial cell model, interleukin (IL)-1beta, mediated a similar ROCK-dependent pattern of tight junction disruption. Further experiments revealed that H. pylori infection induced IL-1 receptor type I (IL-1RI) phosphorylation, independently of epithelial secretion of its endogenous ligands IL-1alpha, IL-1beta or IL-18. Finally, inhibition of IL-1RI activation prevented H. pylori-induced ROCK activation and claudin-4 disruption. Taken together, these findings identify a novel pathophysiological mechanism by which H. pylori disrupts gastric epithelial barrier structure via IL-1RI-dependent activation of ROCK, which in turn mediates tight junctional claudin-4 disruption.
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Células Epiteliales/microbiología , Infecciones por Helicobacter/microbiología , Helicobacter pylori/patogenicidad , Proteínas de la Membrana/antagonistas & inhibidores , Receptores de Interleucina-1/metabolismo , Quinasas Asociadas a rho/metabolismo , Antígenos Bacterianos/fisiología , Proteínas Bacterianas/fisiología , Línea Celular , Claudina-4 , Humanos , Interleucina-1beta/metabolismo , Fosforilación , Factores de Virulencia/fisiologíaRESUMEN
The homeostatic balance of the gastrointestinal tract relies on a single layer of epithelial cells, which assumes both digestive and protective functions. Enteric pathogens, including enteropathogenic Escherichia coli (EPEC), have evolved numerous mechanisms to disrupt basic intestinal epithelial functions, promoting the development of gastrointestinal disorders. Despite its non-invasive nature, EPEC inflicts severe damage to the intestinal mucosa, including the dysregulation of water and solute transport and the disruption of epithelial barrier structure and function. Despite the high prevalence and morbidity of disease caused by EPEC infections, the etiology of its pathogenesis remains incompletely understood. This review integrates the newest findings on EPEC-epithelial interactions with established mechanisms of disease in an attempt to give a comprehensive understanding of the cellular processes whereby this common pathogen may cause diarrheal illness.
Asunto(s)
Diarrea/microbiología , Infecciones por Escherichia coli/microbiología , Escherichia coli/fisiología , Interacciones Huésped-Patógeno/fisiología , Mucosa Intestinal/microbiología , Animales , Diarrea/fisiopatología , Modelos Animales de Enfermedad , Células Epiteliales/metabolismo , Células Epiteliales/microbiología , Escherichia coli/patogenicidad , Infecciones por Escherichia coli/metabolismo , Infecciones por Escherichia coli/fisiopatología , Humanos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/fisiopatología , Transporte Iónico/fisiología , Simportadores/fisiología , Virulencia , Agua/metabolismoRESUMEN
The high efficiency of protein deposition during the neonatal period is driven by high rates of protein synthesis, which are maximally stimulated after feeding. In the current study, we examined the individual roles of amino acids and insulin in the regulation of protein synthesis in peripheral and visceral tissues of the neonate by performing pancreatic glucose-amino acid clamps in overnight-fasted 7-day-old pigs. We infused pigs (n = 8-12/group) with insulin at 0, 10, 22, and 110 ng kg(-0.66) min(-1) to achieve approximately 0, 2, 6 and 30 muU ml(-1) insulin so as to simulate below fasting, fasting, intermediate, and fed insulin levels, respectively. At each insulin dose, amino acids were maintained at the fasting or fed level. In conjunction with the highest insulin dose, amino acids were also allowed to fall below the fasting level. Tissue protein synthesis was measured using a flooding dose of L: -[4-(3)H] phenylalanine. Both insulin and amino acids increased fractional rates of protein synthesis in longissimus dorsi, gastrocnemius, masseter, and diaphragm muscles. Insulin, but not amino acids, increased protein synthesis in the skin. Amino acids, but not insulin, increased protein synthesis in the liver, pancreas, spleen, and lung and tended to increase protein synthesis in the jejunum and kidney. Neither insulin nor amino acids altered protein synthesis in the stomach. The results suggest that the stimulation of protein synthesis by feeding in most tissues of the neonate is regulated by the post-prandial rise in amino acids. However, the feeding-induced stimulation of protein synthesis in skeletal muscles is independently mediated by insulin as well as amino acids.
Asunto(s)
Aminoácidos/metabolismo , Insulina/metabolismo , Músculo Esquelético/metabolismo , Biosíntesis de Proteínas , Porcinos/metabolismo , Vísceras/metabolismo , Animales , Animales Recién Nacidos , Ayuno/metabolismo , Músculo Esquelético/crecimiento & desarrolloRESUMEN
This prospective Canadian pilot study assesses the platelet response rate in H. pylori positive and negative patients and evaluates potential mechanisms to explain response. Patients with ITP received H. pylori eradication therapy and platelet counts at day 56 were used to assess response. Gastric permeability, stool H. pylori antigen and serum CagA anti-body were done at baseline and at day 60. Twenty-two patients were enrolled with an overall response rate of 27% (6/22). The prevalence of H. pylori was 18% (4/22). Seventy-five percent of the H. pylori positive patients (3/4) achieved a response compared to 17% (3/18) of the H. pylori negative patients (P < 0.05). Seventy-five percent of complete responders have demonstrated long-term ongoing responses at 48 months of follow-up. A trend towards lower post-eradication gastric permeability in responders was seen. Although the prevalence of H. pylori is low, H. pylori positive Canadian patients with ITP may benefit from a trial of H. pylori eradication therapy as a safe and effective means to achieve long term platelet response.
Asunto(s)
Antibacterianos/uso terapéutico , Plaquetas/inmunología , Infecciones por Helicobacter/tratamiento farmacológico , Infecciones por Helicobacter/inmunología , Helicobacter pylori , Púrpura Trombocitopénica Idiopática/inmunología , Antígenos Bacterianos/sangre , Antígenos Bacterianos/inmunología , Canadá , Femenino , Infecciones por Helicobacter/epidemiología , Humanos , Masculino , Persona de Mediana Edad , Proyectos Piloto , Púrpura Trombocitopénica Idiopática/epidemiología , Estudios Seroepidemiológicos , Estómago/microbiología , Factores de TiempoRESUMEN
GH treatment increases protein deposition and the efficiency of dietary protein used for growth. To identify the mechanisms that regulate tissue protein synthesis in response to exogenous GH treatment, fully fed, growing swine were treated with GH for 7 d. Fasted and fed pigs were infused with [1-(13)C]leucine to determine protein synthesis rates, and translation initiation factor activity levels were measured in skeletal muscle and liver. Feeding increased protein synthesis and translational efficiency in both muscle and liver of control and GH-treated pigs, and this was associated with increased 4E-BP1 and S6 kinase 1 phosphorylation, decreased association of eukaryotic initiation factor (eIF) 4E with 4E-BP1, and increased association of eIF4E with eIF4G. GH increased muscle protein synthesis and translational efficiency in fed pigs. GH increased liver protein synthesis of fasted and fed pigs in association with increased ribosome number. In muscle, but not liver, GH increased eIF2B activity and 4E-BP1 phosphorylation in both the fasted and fed state and increased the association of eIF4E with eIF4G in the fed state. We conclude that GH increases muscle protein synthesis in the fed state, in part, via mechanisms that enhance the binding of mRNA and methionyl-tRNA to the 40S ribosomal subunit, whereas GH increases liver protein synthesis in the fasted and fed states by increasing ribosome number. The results further indicate that the GH-induced protein synthetic response is dependent upon nutritional state and is tissue specific.
Asunto(s)
Hormona del Crecimiento/farmacología , Hígado/fisiología , Músculo Esquelético/fisiología , Biosíntesis de Proteínas/efectos de los fármacos , Biosíntesis de Proteínas/fisiología , Proteínas Serina-Treonina Quinasas , Animales , Glucemia , Peso Corporal/efectos de los fármacos , Proteínas Portadoras/metabolismo , Ingestión de Alimentos , Factor 2 Eucariótico de Iniciación/metabolismo , Factor 2B Eucariótico de Iniciación/metabolismo , Factor 4E Eucariótico de Iniciación/metabolismo , Factor 4G Eucariótico de Iniciación/metabolismo , Ayuno , Femenino , Glucagón/sangre , Insulina/sangre , Factor I del Crecimiento Similar a la Insulina/metabolismo , Fosfoproteínas/metabolismo , Fosforilación , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Proto-Oncogénicas c-akt , Proteínas Quinasas S6 Ribosómicas/metabolismo , Sus scrofaRESUMEN
Patients with ulcerative colitis and Crohn's disease are at increased risk of developing intestinal cancers via mechanisms that remain incompletely understood. However, chronic inflammation and repeated events of inflammatory relapse in inflammatory bowel disease (IBD) expose these patients to a number of signals known to have tumorigenic effects including persistent activation of the nuclear factor-kappaB and cyclooxygenase-2/prostaglandin pathways, release of proinflammatory mediators such as tumor necrosis factor-alpha and interleukin-6, and enhanced local levels of reactive oxygen and nitrogen species. These inflammatory signals can contribute to carcinogenesis via 3 major processes: 1) by increasing oxidative stress, which promotes DNA mutagenesis thus contributing to tumor initiation; 2) by activating prosurvival and antiapoptotic pathways in epithelial cells, thereby contributing to tumor promotion; and 3) by creating an environment that supports sustained growth, angiogenesis, migration, and invasion of tumor cells, thus supporting tumor progression and metastasis. The present review integrates clinical and basic research observations in an attempt to provide a comprehensive understanding of how inflammatory processes may contribute to intestinal cancer development in IBD patients.
Asunto(s)
Mediadores de Inflamación/metabolismo , Enfermedades Inflamatorias del Intestino/complicaciones , Neoplasias Intestinales/etiología , Animales , Humanos , Enfermedades Inflamatorias del Intestino/inmunología , Neoplasias Intestinales/inmunología , Ratones , Mutación , Estrés Oxidativo , Especies de Nitrógeno Reactivo/metabolismo , Especies Reactivas de Oxígeno/metabolismo , RiesgoRESUMEN
Skeletal muscle protein synthesis is elevated in neonates in part due to an enhanced response to the rise in insulin and amino acids after eating. In vitro studies suggest that glucose plays a role in protein synthesis regulation. To determine whether glucose, independently of insulin and amino acids, is involved in the postprandial rise in skeletal muscle protein synthesis, pancreatic-substrate clamps were performed in neonatal pigs. Insulin secretion was inhibited with somatostatin and insulin was infused to reproduce fasting or fed levels, while glucose and amino acids were clamped at fasting or fed levels. Fractional protein synthesis rates and translational control mechanisms were examined. Raising glucose alone increased protein synthesis in fast-twitch glycolytic muscles but not in other tissues. The response in muscle was associated with increased phosphorylation of protein kinase B (PKB) and enhanced formation of the active eIF4E.eIF4G complex but no change in phosphorylation of AMP-activated protein kinase (AMPK), tuberous sclerosis complex 2 (TSC2), mammalian target of rapamycin (mTOR), 4E-binding protein-1 (4E-BP1), ribosomal protein S6 kinase (S6K1), or eukaryotic elongation factor 2 (eEF2). Raising glucose, insulin, and amino acids increased protein synthesis in most tissues. The response in muscle was associated with phosphorylation of PKB, mTOR, S6K1, and 4E-BP1 and enhanced eIF4E.eIF4G formation. The results suggest that the postprandial rise in glucose, independently of insulin and amino acids, stimulates protein synthesis in neonates, and this response is specific to fast-twitch glycolytic muscle and occurs by AMPK- and mTOR-independent pathways.
Asunto(s)
Glucosa/farmacología , Complejos Multienzimáticos/fisiología , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/metabolismo , Biosíntesis de Proteínas/efectos de los fármacos , Proteínas Quinasas/fisiología , Proteínas Serina-Treonina Quinasas/fisiología , Proteínas Quinasas Activadas por AMP , Aminoácidos/administración & dosificación , Animales , Animales Recién Nacidos , Factores Eucarióticos de Iniciación/metabolismo , Femenino , Glucosa/administración & dosificación , Bombas de Infusión , Insulina/administración & dosificación , Masculino , Transducción de Señal , Porcinos , Serina-Treonina Quinasas TORRESUMEN
Sepsis promotes insulin resistance and reduces protein synthesis in skeletal muscle of adults. The effect of sepsis on insulin-stimulated muscle protein synthesis has not been determined in neonates, a highly anabolic population that is uniquely sensitive to insulin. Overnight fasted neonatal pigs were infused for 8 h with endotoxin [lipopolysaccharide (LPS), 0 and 10 mug.kg(-1).h(-1)]. Glucose and amino acids were maintained at fasting levels, insulin was clamped at either fasting or fed (2 or 10 muU/ml) levels, and fractional protein synthesis rates were determined at the end of the infusion. LPS infusion induced a septic-like state, as indicated by a sustained elevation in body temperature, heart rate, and cortisol. At fasting insulin levels, LPS reduced fractional protein synthesis rates in gastrocnemius muscle (-26%) but had no effect on the masseter and heart. By contrast, LPS stimulated liver protein synthesis (+28%). Increasing insulin to fed levels accelerated protein synthesis rates in gastrocnemius (controls by +38%, LPS by +60%), masseter (controls by +50%, LPS by +43%), heart (controls by +34%, LPS by +40%), and diaphragm (controls by +54%, LPS by +29%), and the response to insulin was similar in LPS and controls. Insulin did not alter protein synthesis in liver, kidney, or jejunum in either group. These findings suggest that acute endotoxemia lowers basal fasting muscle protein synthesis in neonates but does not alter the response of protein synthesis to insulin.
Asunto(s)
Endotoxemia/metabolismo , Insulina/farmacología , Proteínas Musculares/biosíntesis , Músculo Esquelético/metabolismo , Aminoácidos de Cadena Ramificada/sangre , Aminoácidos de Cadena Ramificada/metabolismo , Animales , Animales Recién Nacidos , Glucemia/metabolismo , Endotoxemia/sangre , Femenino , Técnica de Clampeo de la Glucosa , Hidrocortisona/sangre , Insulina/sangre , Lipopolisacáridos , Proteínas Musculares/metabolismo , Músculo Esquelético/fisiopatología , Embarazo , Biosíntesis de Proteínas/efectos de los fármacos , Distribución Aleatoria , PorcinosRESUMEN
Infusion of physiological levels of insulin and/or amino acids reproduces the feeding-induced stimulation of muscle protein synthesis in neonates. To determine whether insulin and amino acids independently stimulate skeletal muscle protein synthesis in neonates, insulin secretion was blocked with somatostatin in fasted 7-day-old pigs (n = 8-12/group) while glucose and glucagon were maintained at fasting levels and insulin was infused to simulate either less than fasting, fasting, intermediate, or fed insulin levels. At each dose of insulin, amino acids were clamped at either the fasting or fed level; at the highest insulin dose, amino acids were also reduced to less than fasting levels. Skeletal muscle protein synthesis was measured using a flooding dose of l-[4-(3)H]phenylalanine. Hyperinsulinemia increased protein synthesis in skeletal muscle during hypoaminoacidemia and euaminoacidemia. Hyperaminoacidemia increased muscle protein synthesis during hypoinsulinemia and euinsulinemia. There was a dose-response effect of both insulin and amino acids on muscle protein synthesis. At each insulin dose, hyperaminoacidemia increased muscle protein synthesis. The effects of insulin and amino acids on muscle protein synthesis were largely additive until maximal rates of protein synthesis were achieved. Amino acids enhanced basal protein synthesis rates but did not enhance the sensitivity or responsiveness of muscle protein synthesis to insulin. The results suggest that insulin and amino acids independently stimulate protein synthesis in skeletal muscle of the neonate.
Asunto(s)
Aminoácidos/farmacología , Animales Recién Nacidos/metabolismo , Insulina/farmacología , Proteínas Musculares/biosíntesis , Músculo Esquelético/efectos de los fármacos , Porcinos/metabolismo , Aminoácidos/sangre , Aminoácidos de Cadena Ramificada/sangre , Animales , Interacciones Farmacológicas , Ayuno , Alimentos , Insulina/sangre , Insulina/metabolismo , Secreción de Insulina , Cinética , Músculo Esquelético/metabolismo , Fenilalanina/administración & dosificación , Somatostatina/farmacología , TritioRESUMEN
The high efficiency of protein deposition during the neonatal period is driven by high rates of protein synthesis, which are maximally stimulated after feeding. Infusion of amino acids, but not insulin, reproduces the feeding-induced stimulation of liver protein synthesis. To determine whether amino acid-stimulated liver protein synthesis is independent of insulin in neonates, and to examine the role of amino acids and insulin in the regulation of translation initiation in neonatal liver, we performed pancreatic glucose-amino acid clamps in overnight-fasted 7-day-old pigs. Pigs (n = 9-12/group) were infused with insulin at 0, 10, 22, and 110 ng.kg(-0.66).min(-1) to achieve 0, 2, 6, and 30 microU/ml insulin, respectively. At each insulin dose, amino acids were maintained at fasting or fed levels or, in conjunction with the highest insulin dose, allowed to fall to below fasting levels. Insulin had no effect on the fractional rate of protein synthesis in liver. Amino acids increased fractional protein synthesis rates in liver at each dose of insulin, including the 0 microU/ml dose. There was a dose-response effect of amino acids on liver protein synthesis. Amino acids and insulin increased protein S6 kinase and 4E-binding protein 1 (4E-BP1) phosphorylation; however, only amino acids decreased formation of the inactive 4E-BPI.eukaryotic initiation factor-4E (eIF4E) complex. The results suggest that amino acids regulate liver protein synthesis in the neonate by modulating the availability of eIF4E for 48S ribosomal complex formation and that this response does not require insulin.
Asunto(s)
Aminoácidos/farmacología , Hipoglucemiantes/farmacología , Insulina/farmacología , Hígado/efectos de los fármacos , Hígado/metabolismo , Factores de Edad , Animales , Animales Recién Nacidos , Glucemia/metabolismo , Factor 4E Eucariótico de Iniciación/metabolismo , Femenino , Técnica de Clampeo de la Glucosa , Hígado/crecimiento & desarrollo , Biosíntesis de Proteínas/efectos de los fármacos , Proteínas Ribosómicas/biosíntesis , Proteínas Ribosómicas/metabolismo , Sus scrofaRESUMEN
Protein synthesis in skeletal muscle is reduced by as much as 50% as early as 4 h after a septic challenge in adults. However, the effect of sepsis on muscle protein synthesis has not been determined in neonates, a highly anabolic population whose muscle protein synthesis rates are elevated and uniquely sensitive to insulin and amino acid stimulation. Neonatal piglets (n = 10/group) were infused for 8 h with endotoxin [lipopolysaccharide (LPS), 0 and 10 microg. kg(-1). h(-1)]. Plasma amino acid and glucose concentrations were kept at the fed level by infusion of dextrose and a balanced amino acid mixture. Fractional protein synthesis rates were determined by use of a flooding dose of [(3)H]phenylalanine. LPS infusion produced a septic-like state, as indicated by an early and sustained elevation in body temperature, heart rate, and plasma tumor necrosis factor-alpha, interleukin-1, cortisol, and lactate concentrations. Plasma levels of insulin increased, whereas glucose and amino acids decreased, suggesting the absence of insulin resistance. LPS significantly reduced protein synthesis in longissimus dorsi muscle by only 11% and in gastrocnemius by only 15%, but it had no significant effect in masseter and cardiac muscles. LPS increased protein synthesis in the liver (22%), spleen (28%), kidney (53%), jejunum (19%), diaphragm (21%), lung (50%), and skin (13%), but not in the stomach, pancreas, or brain. These findings suggest that, when substrate supply is maintained, skeletal muscle protein synthesis in neonates compared with adults is relatively resistant to the catabolic effects of sepsis.
Asunto(s)
Endotoxemia/metabolismo , Proteínas Musculares/biosíntesis , Músculo Esquelético/metabolismo , Factores de Edad , Aminoácidos/metabolismo , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Animales Recién Nacidos , Femenino , Insulina/sangre , Interleucina-1/sangre , Lipopolisacáridos/farmacología , Embarazo , Ribosomas/metabolismo , Sepsis/metabolismo , Porcinos , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Previous studies have shown that intravenous infusion of insulin and/or amino acids reproduces the feeding-induced stimulation of muscle protein synthesis in neonates and that insulin and amino acids act independently to produce this effect. The goal of the present study was to delineate the regulatory roles of insulin and amino acids on muscle protein synthesis in neonates by examining translational control mechanisms, specifically the eukaryotic translation initiation factors (eIFs), which enable coupling of initiator methionyl-tRNAi and mRNA to the 40S ribosomal subunit. Insulin secretion was blocked by somatostatin in fasted 7-day-old pigs (n = 8-12/group), insulin was infused to achieve plasma levels of approximately 0, 2, 6, and 30 microU/ml, and amino acids were clamped at fasting or fed levels or, at the high insulin dose, below fasting. Both insulin and amino acids increased the phosphorylation of ribosomal protein S6 kinase (S6K1) and the eIF4E-binding protein (4E-BP1), decreased the binding of 4E-BP1 to eIF4E, increased eIF4E binding to eIF4G, and increased fractional protein synthesis rates but did not affect eIF2B activity. In the absence of insulin, amino acids had no effect on these translation initiation factors but increased the protein synthesis rates. Raising insulin from below fasting to fasting levels generally did not alter translation initiation factor activity but raised protein synthesis rates. The phosphorylation of S6K1 and 4E-BP1 and the amount of 4E-BP1 bound to eIF4E and eIF4E bound to eIF4G were correlated with insulin level, amino acid level, and protein synthesis rate. Thus insulin and amino acids regulate muscle protein synthesis in skeletal muscle of neonates by modulating the availability of eIF4E for 48S ribosomal complex assembly, although other processes also must be involved.
Asunto(s)
Aminoácidos/farmacología , Animales Recién Nacidos/fisiología , Hipoglucemiantes/farmacología , Insulina/farmacología , Proteínas Musculares/biosíntesis , Músculo Esquelético/metabolismo , Biosíntesis de Proteínas/efectos de los fármacos , Algoritmos , Aminoácidos/metabolismo , Aminoácidos de Cadena Ramificada/metabolismo , Animales , Factor 4E Eucariótico de Iniciación/metabolismo , Factor 4F Eucariótico de Iniciación/biosíntesis , Factor 4F Eucariótico de Iniciación/genética , Femenino , Técnica de Clampeo de la Glucosa , Immunoblotting , Músculo Esquelético/efectos de los fármacos , Pancrelipasa/fisiología , Fosforilación , ARN Mensajero/biosíntesis , ARN Mensajero/genética , PorcinosRESUMEN
Feeding stimulates protein synthesis in skeletal muscle and liver of neonates and this response can be reproduced in muscle by the infusion of insulin or amino acids and in liver by the infusion of amino acids, but not insulin. Activation of insulin signaling components leading to translation initiation is associated with the feeding-induced stimulation of muscle protein synthesis in neonates. In this study, we examined the individual roles of insulin and amino acids in the activation of insulin signaling components leading to translation initiation, specifically, the insulin receptor (IR), insulin receptor substrate 1 (IRS-1), phosphatidylinositol 3-kinase (PI 3-kinase), protein kinase B (PKB) and ribosomal protein S6. Insulin secretion was blocked by somatostatin in food-deprived, 7-d-old pigs (n=8-12/group); insulin was infused to achieve plasma levels of approximately 0, 17, 52, and 255 pmol/L (approximately 0, 2, 6, 30 microU/mL), and amino acids were clamped at food-deprived or fed levels. In skeletal muscle, insulin increased the activation of IR, IRS-1, PI 3-kinase, PKB and S6 and stimulated protein synthesis. In liver, insulin increased the activation of IR, IRS-1, PI 3-kinase, PKB and S6, but had no effect on protein synthesis. Raising amino acids from the food-deprived to the fed level did not alter the insulin-induced activation of IR, IRS-1, PI 3-kinase and PKB but increased S6 phosphorylation and protein synthesis in skeletal muscle and liver. The results suggest that the stimulation of protein synthesis in muscle by insulin involves activation of insulin signaling components, and the stimulation of protein synthesis in muscle and liver by amino acids occurs by mechanisms independent of the early steps of this pathway. Furthermore, amino acids do not alter the insulin-stimulated activation of early steps in the insulin signaling pathway.
Asunto(s)
Aminoácidos/administración & dosificación , Animales Recién Nacidos/metabolismo , Insulina/metabolismo , Insulina/farmacología , Proteínas Serina-Treonina Quinasas , Transducción de Señal/efectos de los fármacos , Porcinos/metabolismo , Aminoácidos/sangre , Animales , Activación Enzimática , Técnica de Clampeo de la Glucosa , Técnicas de Inmunoadsorción , Insulina/sangre , Proteínas Sustrato del Receptor de Insulina , Hígado/metabolismo , Proteínas Musculares/biosíntesis , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Fosfoproteínas/metabolismo , Fosforilación , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Proto-Oncogénicas c-akt , Receptor de Insulina/metabolismo , Proteína S6 Ribosómica/metabolismoRESUMEN
To differentiate the effect of somatotropin (ST) treatment on protein metabolism in the hindquarter (HQ) and portal-drained viscera (PDV), growing swine (n = 20) treated with ST (0 or 150 microg x kg(-1) x day(-1)) for 7 days were infused intravenously with NaH(13)CO(3) and [(2)H(5)]phenylalanine and enterally with [1-(13)C]phenylalanine while in the fed state. Arterial, portal venous, and vena cava whole blood samples, breath samples, and blood flow measurements were obtained for determination of tissue and whole body phenylalanine kinetics under steady-state conditions. In the fed state, ST treatment decreased whole body phenylalanine flux, oxidation, and protein degradation without altering protein synthesis, resulting in an improvement in whole body net protein balance. Blood flow to the HQ (+80%), but not to the PDV, was increased with ST treatment. In the HQ and PDV, ST increased phenylalanine uptake (+44 and +23%, respectively) and protein synthesis (+43 and +41%, respectively), with no effect on protein degradation. In ST-treated and control pigs, phenylalanine was oxidized in the PDV (34-43% of enteral and arterial sources) but not the HQ. In both treatment groups, dietary (40%) rather than arterial (10%) extraction of phenylalanine predominated in gut amino acid metabolism, whereas localized blood flow influenced HQ amino acid metabolism. The results indicate that ST increases protein anabolism in young, growing swine by increasing protein synthesis in the HQ and PDV, with no effect on protein degradation. Differing results between the whole body and the HQ and PDV suggest that the effect of ST treatment on protein metabolism is tissue specific.
Asunto(s)
Hormona del Crecimiento/farmacología , Intestino Delgado/metabolismo , Músculo Esquelético/metabolismo , Biosíntesis de Proteínas , Animales , Peso Corporal/efectos de los fármacos , Isótopos de Carbono , Femenino , Miembro Posterior , Intestino Delgado/efectos de los fármacos , Intestino Delgado/crecimiento & desarrollo , Modelos Biológicos , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/crecimiento & desarrollo , Fenilalanina/farmacocinética , Sistema Porta , Porcinos , Tritio , Tirosina/metabolismo , Vísceras/efectos de los fármacos , Vísceras/crecimiento & desarrollo , Vísceras/metabolismoRESUMEN
In the present study, differential responses of regulatory proteins involved in translation initiation in skeletal muscle and liver during sepsis were studied in neonatal pigs treated with lipopolysaccharide (LPS). LPS did not alter eukaryotic initiation factor (eIF) 2B activity in either tissue. In contrast, binding of eIF4G to eIF4E to form the active mRNA-binding complex was repressed in muscle and enhanced in liver. Phosphorylation of eIF4E-binding protein, 4E-BP1, and ribosomal protein S6 kinase, S6K1, was reduced in muscle during sepsis but increased in liver. Finally, changes in 4E-BP1 and S6K1 phosphorylation were associated with altered phosphorylation of the protein kinase mammalian target of rapamycin (mTOR). Overall, the results suggest that translation initiation in both skeletal muscle and liver is altered during neonatal sepsis by modulation of the mRNA-binding step through changes in mTOR activation. Moreover, the LPS-induced changes in factors that regulate translation initiation are more profound than previously reported changes in global rates of protein synthesis in the neonate. This finding suggests that the initiator methionyl-tRNA-rather than the mRNA-binding step in translation initiation may play a more critical role in maintaining protein synthesis rates in the neonate during sepsis.
Asunto(s)
Lipopolisacáridos/farmacología , Hígado/metabolismo , Músculo Esquelético/metabolismo , Proteínas Quinasas/metabolismo , Animales , Animales Recién Nacidos , Proteínas Portadoras/metabolismo , Factor 4G Eucariótico de Iniciación/metabolismo , Hígado/efectos de los fármacos , Músculo Esquelético/efectos de los fármacos , Fosfoproteínas/metabolismo , Fosforilación , Biosíntesis de Proteínas/efectos de los fármacos , Biosíntesis de Proteínas/fisiología , Proteínas Quinasas/genética , ARN Mensajero/genética , Proteínas Quinasas S6 Ribosómicas/metabolismo , Sepsis/metabolismo , Sepsis/fisiopatología , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Porcinos , Serina-Treonina Quinasas TORRESUMEN
In neonatal pigs, the feeding-induced stimulation of protein synthesis in skeletal muscle, but not liver, can be reproduced by insulin infusion when essential amino acids and glucose are maintained at fasting levels. In the present study, 7- and 26-day-old pigs were studied during 1) fasting, 2) hyperinsulinemic-euglycemic-euaminoacidemic clamps, 3) euinsulinemic-euglycemic-hyperaminoacidemic clamps, and 4) hyperinsulinemic-euglycemic-hyperaminoacidemic clamps. Amino acids were clamped using a new amino acid mixture enriched in nonessential amino acids. Tissue protein synthesis was measured using a flooding dose of L-[4-(3)H]phenylalanine. In 7-day-old pigs, insulin infusion alone increased protein synthesis in various skeletal muscles (from +35 to +64%), with equivalent contribution of myofibrillar and sarcoplasmic proteins, as well as cardiac muscle (+50%), skin (+34%), and spleen (+26%). Amino acid infusion alone increased protein synthesis in skeletal muscles (from +28 to +50%), also with equivalent contribution of myofibrillar and sarcoplasmic proteins, as well as liver (+27%), pancreas (+28%), and kidney (+10%). An elevation of both insulin and amino acids did not have an additive effect. Similar qualitative results were obtained in 26-day-old pigs, but the magnitude of the stimulation of protein synthesis by insulin and/or amino acids was lower. The results suggest that, in the neonate, the stimulation of protein synthesis by feeding is mediated by either amino acids or insulin in most tissues; however, the feeding-induced stimulation of protein synthesis in skeletal muscle is uniquely regulated by both insulin and amino acids.