RESUMO
Our previous studies show that insulin-like growth factor-1 (IGF-1) can either protect against or increase lipopolysaccharide (LPS)-induced damage in the developing brain, depending on the dose, when it is co-administered with LPS through intracerebral injection. To further explore effects of IGF-1 on central inflammation associated brain injury, IGF-1 was administered through intranasal infusion in the current study. Postnatal day 5 (P5) rats were exposed to LPS at a dose of 1 µg/g body weight or sterile saline through intracerebral injection. Recombinant human insulin-like growth factor-1 (rhIGF-1) at a dose of 50 µg/pup or vehicle was administered intranasally 1 or 2 h after the LPS injection. Neonatal LPS exposure resulted in oligodendrocyte (OL) and white matter injury in the P6 or P21 rat brain. The damages include dilatation of lateral ventricles, pyknotic cell death, loss of OL progenitor cells and mature OLs in the cingulum area, and impairment of myelination at the corpus callosum area. Neurological dysfunctions were observed in juvenile rats with neonatal LPS exposure. Intranasal IGF-1 treatment at either 1 or 2 h after LPS exposure significantly attenuated LPS-induced brain injury and improved some behavioral deficits. Intranasal IGF-1 treatment also reduced infiltration of polymorphonuclear (PMN) leukocytes and activation of microglia in the rat brain 24 h after LPS exposure, but it did not prevent the elevation in concentrations of interleukin-1ß (IL-1ß) and tumor necrosis factor alpha (TNFα) in the LPS-exposed rat brain during the first 24 h. This is an indication that direct anti-inflammation might not be the primary mechanism for the protection of IGF-1, and other mechanisms, such as anti-apoptotic effects, are likely involved in its protective effects.
Assuntos
Química Encefálica/fisiologia , Encéfalo/metabolismo , Mediadores da Inflamação/administração & dosagem , Fator de Crescimento Insulin-Like I/administração & dosagem , Lipopolissacarídeos/administração & dosagem , Degeneração Walleriana/fisiopatologia , Degeneração Walleriana/terapia , Administração Intranasal , Animais , Animais Recém-Nascidos , Encéfalo/crescimento & desenvolvimento , Encéfalo/patologia , Modelos Animais de Doenças , Feminino , Humanos , Mediadores da Inflamação/fisiologia , Fator de Crescimento Insulin-Like I/fisiologia , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/toxicidade , Gravidez , Ratos , Ratos Sprague-Dawley , Degeneração Walleriana/patologiaRESUMO
Dexamethasone (Dex) provides neuroprotection against subsequent hypoxia ischemia (HI) in newborn rats, but the mechanism of this neuroprotection is not well understood. It is known that vascular endothelial growth factor A (VEGF) has neuroprotective effects. The objective of this study was to evaluate the role of the VEGF signaling pathway in the Dex-induced neuroprotection in newborn rats. Seven-day-old rat pups had the right carotid artery permanently ligated followed by 140 or 160 min of hypoxia (8% oxygen). Rat pups received two i.p. injections of either saline or Dex (0.25 mg/kg) at 24 and 4 h before HI exposure. To quantify the effects of a glucocorticoid receptor (GR) blocker, on postnatal day (PD) 6 and 15 min prior to Dex treatment rat pups received s.c. vehicle or RU486 (GR blocker, 60 mg/kg). After 24 h at PD 7, all rat pups had HI as described earlier. To quantify the effects of a VEGFR 2 blocker, at 24 h after Dex/Veh treatment (PD7), SU5416, a VEGFR 2 inhibitor or vehicle was injected intracerebroventricularly in the right hemisphere at 30 min before and 2 h after HI. Dex pre-treatment reduced brain injury and enhanced the HI-induced brain VEGF protein while a GR blocker inhibited these effects. Treatment with VEGFR 2 blocker decreased Dex-induced neuroprotection also. Dex pre-treatment enhanced the HI-induced increase in mRNA expression of VEGF splice variants and decreased the HI-induced reduction of Akt phosphorylation. Additionally, it also decreased HI-induced increase of caspase-3 activity and DNA fragments in neonatal rat brain. We conclude that Dex provides robust neuroprotection against subsequent HI in newborn rats via GR likely with the partial involvement of VEGF signaling pathway.
Assuntos
Encéfalo/efeitos dos fármacos , Dexametasona/farmacologia , Hipóxia-Isquemia Encefálica/prevenção & controle , Fármacos Neuroprotetores/farmacologia , Fator A de Crescimento do Endotélio Vascular/biossíntese , Animais , Animais Recém-Nascidos , Western Blotting , Encéfalo/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Hipóxia-Isquemia Encefálica/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para CimaRESUMO
The use of dexamethasone (Dex) in premature infants to prevent and/or treat bronchopulmonary dysplasia can adversely affect early neurodevelopment and probably result in loss of cerebral volume. Vascular endothelial growth factor A (VEGF), specifically VEGF(164) isoform has neurotrophic, neuroprotective and neurogenesis enhancing effects. Previous studies have demonstrated that Dex usually down-regulates VEGF. In the present study we investigated the effect of Dex on brain growth and VEGF in the neonatal rat brain. The pups in each litter were divided into the vehicle (n=84) or Dex-treated (n=98) groups. Rat pups in the Dex group received one of three different regimens of i.p. Dex which included tapering doses on postnatal days 3-6 (0.5, 0.25, 0.125 and 0.06 mg/kg, respectively), or repeated doses of 0.5 or 1 mg/kg/day on postnatal days 4-6 or single dose of 0.031, 0.06, 0.125, 0.25 or 0.5 mg/kg on postnatal day 6. The total VEGF protein and mRNA expression of the three main VEGF splice variants (VEGF(120), VEGF(164), and VEGF(188)) were measured in the rat pup brain using enzyme-linked immunosorbent assay and real-time reverse transcription polymerase chain reaction, respectively. Treatment with Dex significantly decreased the gain of body and brain weight. The tapering and repeated doses of Dex significantly increased caspase-3 activity, VEGF protein and the expression of mRNA of VEGF(164) and VEGF(188) splice variants but the single dose did not. We conclude that Dex is neurodegenerative in the developing brain but also increases VEGF which may play a neurotrophic and neuroprotective role.
Assuntos
Encéfalo/efeitos dos fármacos , Dexametasona/farmacologia , Glucocorticoides/farmacologia , Neurogênese/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Fatores Etários , Animais , Animais Recém-Nascidos , Peso Corporal/efeitos dos fármacos , Caspase 3/metabolismo , Relação Dose-Resposta a Droga , Feminino , Masculino , Tamanho do Órgão/efeitos dos fármacos , Gravidez , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
There are increasing data in support of the hypothesis that inflammatory cytokines are involved in neonatal white matter damage. Despite extensive study of the proinflammatory cytokines tumor necrosis factor-alpha and interleukin-1beta, the role of interleukin-6 in the development of white matter damage is largely unknown. In the present study, the role(s) of interleukin-6 in mediating lipopolysaccharide-induced brain injury and behavioral changes was investigated by the intracerebral injection of lipopolysaccharide with interleukin-6 neutralizing antibody in the 5-day-old rat brain. Brain injury was examined in brain sections at postnatal day 8 and postnatal day 21. Behavioral tests including righting reflex, wire hanging maneuver, cliff avoidance, locomotor activity, gait analysis, responses in the elevated plus-maze and passive avoidance were performed from postnatal day 3 to postnatal day 21. Changes in astroglia, microglia and oligodendrocytes were studied using immunohistochemistry in the postnatal day 21 rat brain. Our results show that interleukin-6 antibody attenuated lipopolysaccharide-induced brain lateral ventricle dilation and improved neurobehavioral performance. Interleukin-6 antibody also suppressed lipopolysaccharide-induced astrogliosis and microglial activation, and increased the number of oligodendrocytes in white matter. However, no changes of tumor necrosis factor-alpha and interleukin-1beta were detected. In contrast, no histopathological changes and glial activation were observed in rats injected with only interleukin-6. The present study indicates that the contribution to brain injury by interleukin-6 depends on its interaction with other lipopolysaccharide-induced agents and not on interleukin-6 alone.
Assuntos
Comportamento Animal/efeitos dos fármacos , Lesões Encefálicas/induzido quimicamente , Lesões Encefálicas/fisiopatologia , Interleucina-6/fisiologia , Lipopolissacarídeos/toxicidade , Fatores Etários , Animais , Animais Recém-Nascidos , Anticorpos/administração & dosagem , Aprendizagem da Esquiva/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Lesões Encefálicas/patologia , Contagem de Células/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Imuno-Histoquímica/métodos , Interleucina-1/metabolismo , Interleucina-6/imunologia , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Atividade Motora/efeitos dos fármacos , Desempenho Psicomotor/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Our previous studies have shown that intracerebral administration of endotoxin, lipopolysaccharide (LPS), induces selective white matter injury and hypomyelination in the neonatal rat brain and that the LPS-induced brain injury is associated with activation of microglia. To test the hypothesis that inhibition of microglial activation may protect against LPS-induced white matter injury, we examined roles of minocycline, a putative suppressor of microglial activation, on LPS-induced brain injury in the neonatal rat. A stereotactic intracerebral injection of LPS (1 mg/kg) was performed in postnatal day 5 Sprague-Dawley rats and control rats were injected with sterile saline. Minocycline (45 mg/kg) was administered intraperitoneally 12 h before and immediately after LPS injection and then every 24 h for 3 days. Inflammatory responses, activation of microglia and brain injury were examined 1 and 3 days after LPS injection. LPS injection resulted in brain injury in selective brain areas, including bilateral ventricular enlargement, cell death at the sub- and periventricular areas, loss of O4+ and O1+ oligodendrocyte (OL) immunoreactivity and hypomyelination, as indicated by decreased myelin basic protein immunostaining, in the neonatal rat brain. Minocycline administration significantly attenuated LPS-induced brain injury in these rat brains. The protective effect of minocycline was associated with suppressed microglial activation as indicated by the decreased number of activated microglial cells following LPS stimulation and with consequently decreased elevation of interleukin 1beta and tumor necrosis factor-alpha concentrations induced by LPS and a reduced number of inducible nitric oxide synthase expressing cells. Protection of minocycline was also linked with the reduction in LPS-induced oxidative stress, as indicated by 4-hydroxynonenal positive OLs. The overall results suggest that reduction in microglial activation may protect the neonatal brain from LPS-induced white matter injury and inhibition of microglial activation might be an effective approach for the therapeutic treatment of infection-induced white matter injury.
Assuntos
Antibacterianos/farmacologia , Encefalopatias/induzido quimicamente , Encefalopatias/prevenção & controle , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/toxicidade , Minociclina/farmacologia , Fármacos Neuroprotetores , Animais , Animais Recém-Nascidos , Encéfalo/crescimento & desenvolvimento , Encefalopatias/patologia , Ventrículos Cerebrais/patologia , Ensaio de Imunoadsorção Enzimática , Feminino , Imuno-Histoquímica , Injeções , Interleucina-1/análise , Interleucina-1/biossíntese , Ativação de Macrófagos/efeitos dos fármacos , Masculino , Microglia/efeitos dos fármacos , Óxido Nítrico Sintase/biossíntese , Óxido Nítrico Sintase Tipo II , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Chronic ischemic brain injuries were studied in 7- and 14-day-old rat pups, which were subjected to bilateral carotid artery occlusion (BCAO) on postnatal day 1. BCAO preferentially injured white matter in the corpus callosum, subcortex and internal capsule areas while largely spared cortical neurons. White matter rarefaction in the corpus callosum was observed in 12 out of the 17 BCAO rat brains and significantly enlarged lateral ventricles were found in five out of seven P14 BCAO rat brains. These white matter changes were similar to injuries found in newborn infants with periventricular leukomalacia (PVL). White matter injuries in the 7-day-old BCAO rat brain were accompanied with increased activation of microglia/macrophages, as indicated by ED1 and OX42 positive immunostaining. Immature oligodendrocytes in the 7-day-old BCAO rat brain, as indicated by O4+/O1+ staining, were much fewer than in the sham-operated rat brain. Immunostaining for myelin basic protein (MBP) at the fimbria hippocampus and the internal capsule areas in the 7-day-old BACO rat brain was also much less than in the control rat brain. Consistent with the immunostaining data, MBP mRNA expression in the 7-day-old, but not in the 14-day-old, BCAO rat brain was significantly less than in the control rat brain. The overall results suggest that pre-oligodendrocytes and immature oligodendrocytes might be major targets for chronic ischemic insults and activated microglia/macrophages are possibly involved in the process of white matter injury.
Assuntos
Isquemia Encefálica/patologia , Encéfalo/patologia , Animais , Animais Recém-Nascidos/fisiologia , Biomarcadores , Encéfalo/metabolismo , Isquemia Encefálica/metabolismo , Doença Crônica , Imuno-Histoquímica/métodos , Macrófagos/patologia , Microglia/patologia , Proteína Básica da Mielina/genética , Proteína Básica da Mielina/metabolismo , Oligodendroglia/patologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Coloração e RotulagemRESUMO
Oligodendrocytes are the primary cells injured in periventricular leukomalacia (PVL), a predominant form of brain white matter lesion in preterm infants. To explore the possible linkage between white matter injury and maternal infection, purified rat O-2A progenitor (Oligodendrocyte-type 2 astrocyte progenitor) cell cultures were used as a model in studying the effects of lipopolysaccharide (LPS), an endotoxin, on survival and differentiation of oligodendrocytes and the involvement of other glial cells in the effects of LPS. O-2A progenitor cells were cultured from optic nerves of 7-day-old rat pups in a chemically defined medium (CDM). Astrocyte and microglia cell cultures were prepared from the cortex of 1-day-old rat brains in the CDM. Direct treatment of LPS (1 microg/ml) to O-2A cells had no effect on viability or differentiation of these cells. When O-2A progenitor cells were cultured in the conditioned medium obtained from either astrocyte or microglial cell cultures for 48 hr, survival rate and differentiation of O-2A cells into mature oligodendrocytes were greatly enhanced as measured by the MTT assay and immunocytochemistry. The conditioned medium obtained from astrocytes or microglia treated with LPS for 48 hr, however, failed to show such a promotional effect on viability and differentiation of O-2A cells. When 5 microg/ml LPS was used to stimulate astrocytes or microglia, the conditioned medium from these glial cell cultures caused O-2A cell injury. The overall results indicate that astrocytes and microglia may promote viability and differentiation of O-2A progenitor cells under physiological conditions, but they may also mediate cytotoxic effects of LPS on oligodendrocytes under an infectious disease biochemical environment.
Assuntos
Astrócitos/efeitos dos fármacos , Leucomalácia Periventricular/fisiopatologia , Lipopolissacarídeos/farmacologia , Microglia/efeitos dos fármacos , Oligodendroglia/efeitos dos fármacos , Células-Tronco/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Astrócitos/metabolismo , Infecções Bacterianas/complicações , Comunicação Celular/efeitos dos fármacos , Comunicação Celular/fisiologia , Morte Celular/efeitos dos fármacos , Morte Celular/fisiologia , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Células Cultivadas/efeitos dos fármacos , Células Cultivadas/metabolismo , Meios de Cultivo Condicionados/farmacologia , Citocinas/efeitos adversos , Endotoxinas/efeitos adversos , Feminino , Humanos , Recém-Nascido , Transmissão Vertical de Doenças Infecciosas , Leucomalácia Periventricular/etiologia , Leucomalácia Periventricular/patologia , Lipopolissacarídeos/metabolismo , Microglia/metabolismo , Oligodendroglia/metabolismo , Gravidez , Efeitos Tardios da Exposição Pré-Natal , Ratos , Ratos Sprague-Dawley , Células-Tronco/citologia , Células-Tronco/metabolismoRESUMO
Induction of proinflammatory cytokines has been proposed to be a link between prenatal maternal intrauterine infection and neonatal brain damage. It is known that the endotoxin, lipopolysaccharide (LPS), released during bacterial infection crosses the placenta. Cytokine induction in the fetal rat brain after maternal administration of LPS was determined by reverse transcriptase-polymerase chain reaction method. LPS suspension in pyrogen-free saline was administered (i.p.) to pregnant rats at 18 d of gestation. The control group was treated with pyrogen-free saline. Expression of the proinflammatory cytokines, tumor necrosis factor-alpha and IL-1beta mRNA, in the fetal rat brain was increased in a dose-dependent manner at 1 h after LPS administration. The great increase in expression of IL-1beta mRNA was only observed at 1 h after injection of LPS (4 mg/kg), whereas the increased expression of tumor necrosis factor-alpha was still detectable from 4 to 24 h after LPS administration. Brain injuries were examined by immunohistochemistry in 8-d-old rat pups born to the dams that were consecutively treated with LPS (500 microg/kg) or pyrogen-free saline on gestation d 18 and 19. No apparent necrotic tissue damage was found in either the LPS group or the control group. Myelin basic protein staining, as a marker of myelin, was clearly observed in the internal capsule and the fimbria hippocampus in the rat brain from the control group. Myelin basic protein staining was much less and weaker in the brains of the LPS-treated group. Glial fibrillary acidic protein-positive astrocytes were observed in both the control and the LPS-treated groups. The LPS-treated group appeared to have more glial fibrillary acidic protein-positive astrocytes in the hippocampal and the cortex areas of the brain than the control group. Immunoblotting data showed that glial fibrillary acidic protein content in the cortex or the hippocampus of the LPS-treated rat brain was higher than in the control group. OX-42-positive staining (a marker of the type 3 complement receptors) of microglial cells was greatly reduced in the 8-d-old rat brain after maternal LPS administration. However, histochemistry with tomato lectin showed that staining of both amoeboid and ramified microglial cells in the LPS-treated rat brain was similar to that in the control group. The overall results indicate that maternal LPS administration induces an increased expression of IL-1beta and tumor necrosis factor-alpha mRNA in the fetal brain. Maternal LPS administration also increases glial fibrillary acidic protein-positive astrocytes, decreases myelin basic protein and alters immunoreactivity of microglia in the brain of offspring. Although results from the current study do not provide direct evidence linking LPS-induced cytokines with the abnormalities in the neonatal rat brain, our animal model may be appropriate for exploring the mechanisms involved in the effects of maternal infection on glial cells in the brains of offspring.
Assuntos
Encéfalo/efeitos dos fármacos , Interleucina-1/biossíntese , Lipopolissacarídeos/farmacologia , Fator de Necrose Tumoral alfa/biossíntese , Animais , Sequência de Bases , Western Blotting , Encéfalo/embriologia , Encéfalo/metabolismo , Primers do DNA , Feminino , Feto/efeitos dos fármacos , Proteína Glial Fibrilar Ácida/metabolismo , Imuno-Histoquímica , Interleucina-1/genética , Gravidez , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Taxa de Sobrevida , Fator de Necrose Tumoral alfa/genéticaRESUMO
The effects of intrauterine hypoxia-ischemia (HI) on nitric oxide synthase (NOS) activity and on expression of NOS isoforms were investigated in fetal and neonatal rat brains. Rat fetuses were subjected to either a 30-min intrauterine HI insult or a sham operation (SH) on gestational day 17 (G17). NOS activity in the homogenate of the rat brain was detectable on G17 and increased with age. NOS activity in the HI group was 20-30% higher than in the SH group from 6 to 48 h after the HI, but was 30% lower than in the SH group from postnatal day 8 to 14. Expression of the inducible NOS (iNOS) mRNA, as examined by RT-PCR, was increased as compared to the SH group from 6 to 24 h after the HI surgery. Expression of the constitutive neuronal NOS (nNOS) mRNA was reduced in the HI group from 24 h after the HI surgery up to postnatal day 14. Immunoblotting data have shown that alterations in NOS isoform protein expression caused by the intrauterine HI were consistent with the mRNA expression data. The overall results indicate that prenatal HI has long-lasting effects on function and expression of NOS in fetal and neonatal rat brains and that the altered NOS activity may be associated with prenatal HI-induced neurological abnormalities.
Assuntos
Química Encefálica/fisiologia , Encéfalo/crescimento & desenvolvimento , Hipóxia Encefálica/enzimologia , Ataque Isquêmico Transitório/enzimologia , Proteínas do Tecido Nervoso/biossíntese , Óxido Nítrico Sintase/biossíntese , Animais , Encéfalo/embriologia , DNA/biossíntese , Proteínas do Tecido Nervoso/genética , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase Tipo I , Reação em Cadeia da Polimerase , RNA Mensageiro/biossíntese , RatosRESUMO
Effects of intrauterine hypoxia-ischemia (HI) on receptor-stimulated phosphoinositide (PPI) hydrolysis were studied in rat cerebellar granule cell cultures prepared from an in utero HI model. On gestation day 17, HI conditions were achieved by complete clamping of the uterine vasculature for 30 min followed by removal of the clamps to permit reperfusion. Sham operation (SH, surgery without vasculature ligation) was performed as the control. Intrauterine HI did not affect the basal level of PPI hydrolysis (in the absence of stimulants) in cells prepared from either the SH or the HI group. PPI hydrolysis stimulated by quisqualate (QA) or trans-(1S,3R)-1-amino-1,3-cyclo-pentanedicarboxylic acid (trans-ACPD) was significantly reduced in cells prepared from the HI group, whereas intrauterine HI did not affect the PPI hydrolysis induced by ionotropic glutamate receptor agonists or by norepinephrine or serotonin. At a dose range of 100-300 microM, QA-stimulated PPI hydrolysis in cells prepared from the SH group increased by 3-to 4.5-fold, while this increase was only 2- to 2.5-fold in cells prepared from the HI group. Presence of L-NG-monomethyl-arginine (L-NMMA), a nitric oxide (NO) synthase inhibitor, did not increase QA-stimulated PPI hydrolysis in cells prepared from either the SH or the HI group, indicating that stimulation of NO formation is unlikely involved in the suppressive effects of intrauterine HI on QA-induced PPI hydrolysis. The QA-stimulated PPI hydrolysis in cells prepared from the HI group, but not from the SH group, was further inhibited by L-(+)-2-amino-3-phosphono-propionic acid (L-AP3). The overall results suggest that intrauterine HI has long-lasting suppressive effects on metabotropic glutamate receptor agonist-stimulated PPI hydrolysis and these effects might be associated with alterations in expression of metabotropic glutamate receptor subtypes.
Assuntos
Isquemia Encefálica/fisiopatologia , Cerebelo/citologia , Hipóxia Fetal/fisiopatologia , Hipóxia Encefálica/fisiopatologia , Fosfatidilinositóis/metabolismo , Receptores de Glutamato Metabotrópico/agonistas , Alanina/análogos & derivados , Alanina/farmacologia , Animais , Isquemia Encefálica/complicações , Contagem de Células , Células Cultivadas/química , Células Cultivadas/metabolismo , Cerebelo/irrigação sanguínea , Cicloleucina/análogos & derivados , Cicloleucina/farmacologia , Inibidores Enzimáticos/farmacologia , Agonistas de Aminoácidos Excitatórios/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Feminino , Hidrólise , Hipóxia Encefálica/complicações , Neurônios/química , Neurônios/metabolismo , Neurotoxinas/farmacologia , Gravidez , Proteínas/análise , Ácido Quisquálico/farmacologia , Ratos , Ratos Sprague-Dawley , ômega-N-Metilarginina/farmacologiaRESUMO
Perinatal hypoxic-ischemic (HI) insult is known to cause cellular and molecular disturbances leading to functional and behavioral abnormalities during brain development. In this study, we examined the effects of an in utero HI insult on poly-phosphoinositide turnover in vivo in the cerebrum and cerebellum as well as cholinergic-stimulated turnover in cortical slices from developing rat brain. In utero HI treatment was carried out by clamping the uterine blood vessels of near-term fetuses for 5, 10 and 15 min followed by resuscitation of the newborn pups. The in vivo protocol for examining poly-PI signaling activity in 2 week-old pup brain involved intracerebral injection of [3H]inositol for 16 hr and subsequent intraperitoneal injection with lithium (8 meq/kg) for 4 hr prior to decapitation. In the control pups, lithium elicited a 2.6 fold increase in labeled inositol phosphate (IP) in the cerebrum as compared to a 1.3 fold increase in the cerebellum. In utero HI insult (5 to 15 min) resulted in a small increase in labeled IP in the cerebrum but not in the cerebellum. Carbachol stimulation of poly-PI turnover was examined in brain slices prelabeled with [3H]inositol in vivo. Incubation of the prelabeled slices with carbachol in the presence of LiCl (10 mM) resulted in a time-, dose- and age-dependent increase in labeled IP. Brain slices from 2 week-old pups that experienced in utero HI-treatment for 10 and 15 min (but not 5 min) showed a significant decrease in carbachol-stimulation of labeled IP as compared with control pups. These results indicate the effects of in utero HI on the choninergic-stimulated poly-PI signaling pathway and its implication on related functional deficits in the developing brain.
Assuntos
Encéfalo/crescimento & desenvolvimento , Encéfalo/metabolismo , Hipóxia/metabolismo , Isquemia/metabolismo , Fosfatos de Fosfatidilinositol/metabolismo , Útero/irrigação sanguínea , Animais , Encéfalo/embriologia , Carbacol/farmacologia , Constrição , Feminino , Inositol/metabolismo , Cloreto de Lítio/farmacologia , Parassimpatomiméticos/farmacologia , Gravidez , Ácido Quisquálico/farmacologia , Ratos , Ratos Sprague-Dawley , Cloreto de Sódio/farmacologia , TrítioRESUMO
Effects of intrauterine hypoxia-ischemia (HI) on brain functional development in the term rat were examined in cerebellar granule cell cultures obtained from an in utero HI model. On gestation d 17, HI conditions were achieved by complete clamping of the uterine vasculature for designated durations followed by removal of the clamps to permit reperfusion. Sham operation (surgery without vasculature clamping) was performed as the control. After surgery, the uterine horns were returned to dam's abdomen to let the pups deliver naturally. Severe HI insult from the surgical manipulation was evident in that the lactate levels of fetal brain increased, and fetal blood pH decreased with the duration of vasculature clamping up to 1 h. The experimental HI insult up to 1 h did not affect fetal survival rate, but retarded growth of fetuses or newborns was observed in the 1 h HI group. N-Methyl-D-aspartate (NMDA)- and kainate (KA)-stimulated cGMP formation and glutamate accumulation were measured in cerebellar granule cell cultures from 8-d-old pups suffering from various durations of antenatal HI insult. NMDA (100 microM)-induced elevation of cGMP was further augmented by a 10-35-min HI insult as compared with the control cells (62.4-78.2 versus 49 pmol/mg protein). In the presence of L-NG-monomethyl-arginine (L-NMMA, 150 microM), a nitric oxide synthase inhibitor, NMDA-induced cGMP formation was blocked, and the blockade of cGMP formation by L-NMMA (10 microM) could be reversed by simultaneous application of 1 mM arginine in both control and HI cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Isquemia Encefálica/metabolismo , Doenças Cerebelares/metabolismo , GMP Cíclico/biossíntese , Agonistas de Aminoácidos Excitatórios/farmacologia , Hipóxia Fetal/metabolismo , Hipóxia Encefálica/metabolismo , Animais , Arginina/análogos & derivados , Arginina/farmacologia , Isquemia Encefálica/complicações , Células Cultivadas , Doença Crônica , Inibidores Enzimáticos/farmacologia , Idade Gestacional , Ácido Glutâmico/metabolismo , Hipóxia Encefálica/complicações , Ácido Caínico/farmacologia , N-Metilaspartato/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Ratos , Ratos Sprague-Dawley , ômega-N-MetilargininaRESUMO
Prenatal ethanol exposure-induced alteration in poly-phosphoinositide (PPI) hydrolysis stimulated by excitatory amino acids (EAA) was studied in rat cerebellar granule cells previously labeled with [3H]myoinositol. The prenatal exposure to ethanol was achieved via maternal consumption of a Sustacal (chocolate flavored) liquid diet containing either 5% ethanol (w/v, 35% of calories) or isocaloric sucrose (pair-fed) substituted for ethanol from gestation d 11 until the day of parturition. The ionotropic glutamate receptor agonists, N-methyl-D-aspartate, kainate or (+/-)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) (100 microM each) induced a two- to four-fold increase in PPI hydrolysis over the basal level, regardless of the liquid dietary treatment. Stimulation with quisqualate (QA), an agonist activating both metabotropic and ionotropic glutamate receptors, resulted in a much stronger and dose-dependent response in PPI hydrolysis and exposure in utero to ethanol significantly reduced this response. Tetrodotoxin, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), or (+/-)-3-(2-carboxypiperazine-4-yl)-propyl-1-phosphonic acid (CPP) had no effect on QA-stimulated PPI hydrolysis nor on the suppression of this hydrolysis by ethanol. Exposure in utero to ethanol did not affect PPI hydrolysis stimulated by a selective metabotropic glutamate receptor agonist, trans-(+/-)-l-amino-1,3-cyclopentanedicarboxylic acid (t-ACPD). Although the PPI hydrolysis stimulated by t-ACPD could be blocked by (RS)-alpha-methyl-4-carboxyphenylglycine (MCPG), an antagonist of the metabotropic glutamate receptor, MCPG was incapable of affecting QA-induced PPI hydrolysis and the suppressive effects of prenatal ethanol exposure on this hydrolysis. Taken together, the data suggest that the long-lasting suppressive effects of prenatal ethanol exposure on QA-stimulated PPI hydrolysis in cerebellar granule cell cultures is through a metabotropic QA receptor pathway that may be different from the one activated by t-ACPD.
Assuntos
Cerebelo/metabolismo , Etanol/farmacologia , Fosfatos de Fosfatidilinositol/metabolismo , Diester Fosfórico Hidrolases/metabolismo , Efeitos Tardios da Exposição Pré-Natal , Ácido Quisquálico/farmacologia , Animais , Células Cultivadas , Cerebelo/efeitos dos fármacos , Cerebelo/enzimologia , Etanol/administração & dosagem , Agonistas de Aminoácidos Excitatórios/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Feminino , Inositol/metabolismo , Gravidez , Ratos , Receptores de Glutamato Metabotrópico/agonistas , Receptores de Glutamato Metabotrópico/antagonistas & inibidores , Receptores de Glutamato Metabotrópico/efeitos dos fármacosRESUMO
Docosahexaenoic acid (DHA; 22:6n-3) is important for normal visual development. We hypothesized that preterm infants fed formulas with marine oil as a source of DHA would have better visual acuity than infants fed formulas without marine oil, as measured by the Teller Acuity Card procedure. Marine oil (P < 0.001) and age (P < 0.0001) influenced visual acuity, by repeated-measures analysis of variance (ANOVA) corrected for the effect of subject. Marine-oil-supplemented infants had better visual acuity than those fed standard formulas at 2 and 4 mo of age, by Fishers' least-squares difference (LSD). Acuity of both dietary groups improved through 6.5 mo of age, then plateaued. Through 4 mo of age, acuity was inversely related to oxygen supplementation (log10 h) and positively related to DHA status, by general-linear-models (GLM) analysis. After 4 mo of age, birth weight and gestational age were the only variables consistently related to visual acuity by GLM. We conclude that marine-oil-supplemented formula improved visual acuity of preterm infants through 4 mo of age by improving DHA status.
Assuntos
Óleos de Peixe/farmacologia , Recém-Nascido Prematuro/fisiologia , Acuidade Visual , Envelhecimento/fisiologia , Ácidos Docosa-Hexaenoicos/sangue , Óleos de Peixe/administração & dosagem , Humanos , Recém-Nascido , Recém-Nascido Prematuro/sangue , Modelos Lineares , Ácidos Linolênicos/sangueRESUMO
Red blood cell (RBC) phospholipids of infants fed human milk compared with formula have more arachidonic acid (AA) and docosahexanoic acid (DHA). The addition of low levels of marine oil to infant formula with 0.6 to 2.0% alpha-linolenic acid (LLA, 18:3n-3) prevented declines in DHA in formula-fed infants; however, the feeding trials were short (4 to 6 wk), LLA concentrations were low compared with current formulas (3.0 to 5.0% LLA), and the formulas were unstable. Trials with stable formulas were necessary to determine if dietary DHA could maintain phospholipid DHA after discharge from the hospital and, in fact, if it was necessary with higher intakes of LLA. The results of acute (4 wk) and extended (to 79 wk postconception) feeding of such formulas on RBC and plasma phospholipid AA and DHA are reported here. Control formulas were identical to commercially available formulas. Experimental formulas differed only in the addition of small amounts of marine oil. DHA in RBC and plasma phosphatidylethanolamine (PE) declined during four weeks of feeding but not if marine oil provided DHA (0.2% or 0.4%) and plasma phospholipid AA (g/100 g) decreased with time and marine oil feeding. Extended feeding with marine oil accounted for half the DHA in RBC and plasma phosphatidylethanolamine at equilibrium; however, RBC (g/100 g) and plasma AA (g/100 g; mg/L plasma) decreased progressively until late infancy and were depressed further by marine oil.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Gorduras Insaturadas na Dieta/administração & dosagem , Óleos de Peixe/administração & dosagem , Alimentos Infantis , Fosfolipídeos/sangue , Ácido Araquidônico/sangue , Ácidos Docosa-Hexaenoicos/administração & dosagem , Ácidos Docosa-Hexaenoicos/sangue , Ácido Eicosapentaenoico/sangue , Eritrócitos/metabolismo , Humanos , Fenômenos Fisiológicos da Nutrição do Lactente , Recém-Nascido de Baixo Peso , Recém-Nascido , Recém-Nascido Prematuro , Ácidos Linolênicos/administração & dosagem , Estado Nutricional , Ácido alfa-LinolênicoRESUMO
Indomethacin has proven effective in closing the patent ductus arteriosus (PDA) in most low birth weight (LBW) neonates with this disorder. Early reopening of the ductus is a problem and often leads to the need for surgery. Prolonged use of indomethacin for several days has been suggested as a means to alleviate this problem. The present study was designed to determine if prolonged therapy over 5 days is more effective than a two-dose regimen in preventing reopening of the PDA. Seventy neonates were randomized for either prolonged therapy over 1 week or to receive two doses of indomethacin. All infants were given two doses of indomethacin 0.15 mg per kg, 12h apart. The maintenance group received an additional 0.1 mg per kg daily for 5 days. Ten days after the infants' initial dose of indomethacin, 6 of 22 in the nonmaintenance group as compared to 0 of 22 in the maintenance group had reopening of their ductus arteriosus. Ten infants in the maintenance group eventually had the ductus reopen at a median of 29, range 11-66 days compared to a median of 3, range 2-44 days in the nonmaintenance group. Significantly fewer babies in the maintenance group had a grade II-IV intraventricular hemorrhage compared to the nonmaintenance group. There was no other significant difference in the two groups in the incidence of necrotizing enterocolitis, retrolental fibroplasia or death. Indomethacin given over 5 days is effective for closure of the ductus arteriosus and will prevent reopening until after the acute clinical course in babies under 1500 g; however, the overall incidence of reopening was not different.
Assuntos
Permeabilidade do Canal Arterial/tratamento farmacológico , Indometacina/administração & dosagem , Recém-Nascido de Baixo Peso , Hemorragia Cerebral/complicações , Esquema de Medicação , Permeabilidade do Canal Arterial/complicações , Permeabilidade do Canal Arterial/patologia , Feminino , Humanos , Indometacina/uso terapêutico , Recém-Nascido , Masculino , Fatores de TempoRESUMO
In ascorbic acid-requiring species (human, guinea pig), elevations of circulating histamine occur as the result of marginal ascorbic acid status. Marginal ascorbic acid status during pregnancy is associated with preeclampsia, abruption, and prematurity. Furthermore, circulating histamine is known to be elevated in these complications perhaps as a result of placental dysfunction which diminishes normal placental histaminase. We hypothesized that women with preeclampsia and premature labor would have elevated histamine and the lowest concentrations of ascorbic acid. Plasma total whole blood histamine and ascorbic acid were surveyed in women in term (T) and preterm (PT) labor. Blood histamine was elevated in PT compared to T labor but so was plasma ascorbate, indicating that marginal ascorbate status does not cause the elevated circulating histamine observed in PT. However, marginal ascorbate status concomitant with reduced placental histaminase may contribute to further increases in circulating histamine and to any pathology which might result from elevated histamine. Regression analysis of histamine on ascorbate for T and PT labor revealed a significant inverse relationship between ascorbate and histamine only in PT labor (p less than 0.027). An unexpected finding was that a history of maternal cigarette smoking, to a degree which resulted in marginal ascorbic acid status, confounded the relationship between ascorbate and circulating histamine in T labor.
Assuntos
Ácido Ascórbico/sangue , Histamina/sangue , Trabalho de Parto , Gravidez/sangue , Adulto , Deficiência de Ácido Ascórbico/sangue , Feminino , Humanos , Fumar/efeitos adversosRESUMO
Very low birth weight infants demonstrate significant reductions in red blood cell membrane docosahexaenoic acid (DHA, 22:6n-3) following delivery unless fed human milk. The purpose of the present study was to determine if a dietary source of DHA (MaxEPA, R. P. Scherer Corporation, Troy, MI) could prevent the decline in red blood cell phospholipid DHA in very low birth weight infants whose enteral feeding consisted of a preterm formula without DHA. Longitudinal data were obtained on membrane phospholipid DHA in both unsupplemented and MaxEPA-supplemented infants by a combination of thin-layer and gas chromatography. These infants (n = 39) ranged in age from 10 to 53 days at enrollment (0 time). At enrollment, phospholipid DHA and arachidonic acid (20:4n-6) were inversely correlated with age in days. During the study, mean red blood cell phospholipid DHA declined without supplementary DHA as determined by biweekly measurement, but infants supplemented with MaxEPA maintained the same weight percent of phospholipid (phosphatidylethanolamine, phosphatidylcholine, and phosphatidylserine) DHA as at enrollment. The pattern of red blood cell phospholipid fatty acids in supplemented infants was similar to that reported for preterm infants fed human milk.
Assuntos
Ácido Eicosapentaenoico , Membrana Eritrocítica/efeitos dos fármacos , Ácidos Graxos Insaturados/sangue , Ácidos Graxos Insaturados/uso terapêutico , Óleos de Peixe/uso terapêutico , Ensaios Clínicos como Assunto , Gorduras Insaturadas na Dieta/uso terapêutico , Ácidos Docosa-Hexaenoicos , Combinação de Medicamentos , Membrana Eritrocítica/metabolismo , Ácidos Graxos/sangue , Humanos , Recém-Nascido , Recém-Nascido Prematuro/sangue , Fosfolipídeos/sangueAssuntos
Displasia Broncopulmonar/prevenção & controle , Doença da Membrana Hialina/terapia , Pneumotórax/prevenção & controle , Respiração Artificial , Displasia Broncopulmonar/etiologia , Humanos , Doença da Membrana Hialina/complicações , Doença da Membrana Hialina/mortalidade , Recém-Nascido , Pneumotórax/etiologia , Respiração Artificial/efeitos adversos , Respiração Artificial/métodosRESUMO
Chronic lung disease in the form of stage III or IV bronchopulmonary dysplasia (BPD) was found to occur among nine of 188 infants receiving intermittent positive-pressure ventilation for respiratory insufficiency. None occurred in infants ventilated with a mask alone. The duration of endotracheal intubation was not significantly longer in patients developing BPD; however, the duration of exposure to inspired oxygen greater than 60 percent was significantly longer in that group (mean, 141 hours). A previously undescribed form of chronic lung disease was found to occur among nine of 150 infants ventilated because of underlying idiopathic respiratory distress syndrome. A radiographic appearance of diffuse haziness with loss of identifiable lung markings occurred at 5 to 15 days of age and was not associated with clinical signs or symptoms or increased oxygen needs. These changes disappeared in one to five days.