RESUMO
Na(+)/K(+)-ATPase is a membrane-bound enzyme responsible for Na(+)/K(+) translocation across cell membranes. It is essential for the generation of electrochemical gradients, which control the ionic environment necessary for electrical activity and water and electrolyte balance. Newborn infants who are at risk of developing bronchopulmonary dysplasia (BPD) are frequently treated with corticosteroids. Although these infants are at risk for neurological, water and electrolyte abnormalities, there is little information regarding the effects of clinically relevant doses of corticosteroids on Na(+)/K(+)-ATPase activity and protein isoform expression in the brain and kidney of newborns. In the present study, we examined the effects of dexamethasone on cerebral cortical and renal cortical Na(+)/K(+)-ATPase activity and alpha1- and beta1-protein isoform expression in newborn lambs. Lambs were given four injections of a placebo (n = 11) or one of three different doses of dexamethasone (0.01 mg kg(-1), n = 9; 0.25 mg kg(-1), n = 11; or 0.50 mg kg(-1), n = 9) 12 h apart on Postnatal Days 3 and 4 up to 18 h before harvest of the cerebral cortex and renal cortex. We selected doses in a range to approximate those used to treat infants with BPD. Na(+)/K(+)-ATPase activity was measured in membrane preparations as ouabain-sensitive inorganic phosphate liberation from ATP and alpha1- and beta1-subunit abundance by Western immunoblot. Postnatal treatment of lambs with dexamethasone resulted in a 21.4% increase in Na(+)/K(+)-ATPase activity and a 30.4% increase in catalytic alpha1-protein expression in the cerebral cortex at a dose of 0.50 mg kg(-1) dexamethasone, but not at the lower doses. Dexamethasone treatment was not associated with changes in beta1-isoform expression in the cerebral cortex. In the kidney, dexamethasone treatment was not associated with significant changes in Na(+)/K(+)-ATPase activity or alpha1- or beta1-isoform expression for the doses we examined. Therefore, clinically relevant corticosteroid treatment exerts dose-related, differential organ-specific effects on Na(+)/K(+)-ATPase activity and protein isoform expression in newborn lambs.
Assuntos
Corticosteroides/administração & dosagem , Animais Recém-Nascidos , Córtex Cerebral/enzimologia , Córtex Renal/enzimologia , Ovinos , ATPase Trocadora de Sódio-Potássio/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Western Blotting , Peso Corporal , Dexametasona/administração & dosagem , Relação Dose-Resposta a Droga , Especificidade de Órgãos , Placebos , Subunidades Proteicas/análiseRESUMO
The aim of the present study was to determine the ontogeny and effects of corticosteroid pretreatment on aquaporin 4 (AQP4) channel mRNA and protein expression in the cerebral cortex of sheep during development. A portion of the cerebral cortex was snap-frozen from fetuses of dexamethasone- and placebo-treated ewes at 60%, 80% and 90% of gestation, dexamethasone- and placebo-treated newborn lambs and adult sheep. Cerebral cortical samples were obtained 18 h after the last of four 6 mg dexamethasone or placebo injections were given over 48 h to the ewes and adult sheep. Lambs were treated with 0.01 mg kg(-1) dexamethasone or placebo in the same schedule as the ewes and adult sheep. Amplification of an ovine AQP4 cDNA fragment was accomplished by reverse transcription-polymerase chain reaction using primers based on a homologous bovine sequence. The resulting cDNA was used to determine AQP4 channel mRNA expression by Northern hybridisation using phosphorimaging. The relative abundance of AQP4 mRNA was normalised to the ovine ribosomal gene L32. A portion of the frontal cortex was also analysed for AQP4 protein expression by Western immunoblot. Densitometry was performed and the results expressed as a ratio to an adult brain pool. Aquaporin 4 channel mRNA and protein were detectable as early as at 60% gestation. There were no changes in AQP4 mRNA expression among the fetal, newborn and adult groups or after dexamethasone pretreatment in any age group. The expression of the AQP4 protein was higher (P < 0.05) in fetuses at 80% and 90% of gestation (2.9- and 3.3-fold, respectively), in lambs (3.2-fold) and in adult sheep (3.8-fold) compared with fetuses at 60% of gestation, as well as in adult sheep (1.3-fold) compared with fetuses at 80% of gestation. Dexamethasone pretreatment resulted in decreases (P < 0.05) in AQP4 protein expression in the lambs and adult sheep, but not in the fetal groups. We conclude that: (1) AQP4 mRNA and protein were expressed early in fetal and throughout ovine development; (2) protein, but not mRNA, expression increased between 60% and 80% of gestation and did not differ from adult levels by 90% of gestation; and (3) dexamethasone pretreatment resulted in decreases in AQP4 protein expression in lambs and adult sheep, but not in fetuses. The maturational increases in AQP4 protein expression and dexamethasone-related decreases in expression were post-transcriptional, because changes in AQP4 mRNA expression were not observed.
Assuntos
Corticosteroides/administração & dosagem , Aquaporina 4/efeitos dos fármacos , Córtex Cerebral/química , Córtex Cerebral/embriologia , Desenvolvimento Fetal , Ovinos/embriologia , Animais , Aquaporina 4/análise , Aquaporina 4/genética , Western Blotting , Dexametasona/administração & dosagem , Idade Gestacional , Hidrocortisona/sangue , RNA Mensageiro/análiseRESUMO
The two extremes of abnormal fetal growth are restricted growth and excessive growth, both of which originate from alterations in the uterine metabolic milieu. The fetus must adapt to these conditions to survive. In both instances, however, the inciting insult and the subsequent adaptation of the fetus carry long-term health consequences. In some instances, these changes may have generational implications. Counseling and care by pediatricians should be directed at the continuum of age ranges, including the expectant mother, the newborn, the child and adolescent, and future generations.
Assuntos
Peso ao Nascer , Desenvolvimento Embrionário e Fetal/fisiologia , Retardo do Crescimento Fetal/etiologia , Retardo do Crescimento Fetal/fisiopatologia , Macrossomia Fetal/etiologia , Macrossomia Fetal/fisiopatologia , Humanos , Recém-Nascido , Recém-Nascido Pequeno para a Idade Gestacional , Fatores de RiscoRESUMO
This study examined the utility of serum uric acid concentrations in the first day of life to identify infants with severe brain injury (grade III or IV intraventricular hemorrhage and/or periventricular leukomalacia). The serum uric acid concentrations in infants with severe brain injury were compared to those without. Severe brain injury was assessed in 151 infants with birthweight < or = 1,251 g admitted before 24 h of life. The risk of severe brain injury was related to 5-min Apgar scores (odds ratio 0.79, CI 0.63-0.98, p < 0.05) and seizures in the first day of life (odds ratio 4.44, CI 1.004-19.66, p < 0.05), but the mean uric acid levels did not differ between those with and without severe brain injury [5.11 +/- 1.88 mg/dl (303.9 +/- 111.8 micromol/l) vs. 5.77 +/- 2.13 mg/dl (343.2 +/- 126.7 micromol/l), p = 0.200]. Uric acid levels were related to serum creatinine (p < 0.001), time of uric acid sample (p < 0.001), maternal hypertension (p < 0.001), and base deficit (p = 0.032). Of the 80 infants seen in neurodevelopmental follow-up at a median 11 months postconceptional age, uric acid concentrations did not differ between the abnormal (n = 20) and normal subjects [5.38 +/- 1.72 mg/dl (320.0 +/- 102.3 micromol/l) vs. 6.02 +/- 2.55 mg/dl (358.1 +/- 151.8 micromol/l), p = 0.197]. Uric acid may not be a useful early marker for premature infants with severe brain injury.