RESUMO
BACKGROUND: Efficacy of surfactant therapy in fetal growth restricted (FGR) preterm neonates is unknown. METHODS: Twin-bearing ewes underwent surgery at 105 days gestation to induce FGR in one twin by single umbilical artery ligation. At 123-127 days, catheters and flow probes were implanted in pulmonary and carotid arteries to measure flow and pressure. Lambs were delivered, intubated and mechanically ventilated. At 10 min, surfactant (100 mg kg-1) was administered. Ventilation, oxygenation, and hemodynamic responses were recorded for 1 h before euthanasia at 120 min. Lung tissue and bronchoalveolar lavage fluid was collected for analysis of surfactant protein mRNA and phosphatidylcholines (PCs). RESULTS: FGR preterm lambs were 26% lighter than appropriate for gestational age (AGA) lambs and had baseline differences in lung mechanics and pulmonary blood flows. Surfactant therapy reduced ventilator and oxygen requirements and improved lung mechanics in both groups, although a more rapid improvement in compliance and tidal volume was observed in AGA lambs. Surfactant administration was associated with decreased mean pulmonary and carotid blood flow in FGR but not AGA lambs. No major differences in surfactant protein mRNA or PC levels were noted. CONCLUSIONS: Surfactant therapy was associated with an altered pulmonary and cerebral hemodynamic response in preterm FGR lambs.
Assuntos
Retardo do Crescimento Fetal/fisiopatologia , Hemodinâmica/efeitos dos fármacos , Pulmão/metabolismo , Surfactantes Pulmonares/uso terapêutico , Animais , Animais Recém-Nascidos , Encéfalo/efeitos dos fármacos , Líquido da Lavagem Broncoalveolar , Retardo do Crescimento Fetal/metabolismo , Coração/efeitos dos fármacos , Coração/fisiopatologia , Pulmão/efeitos dos fármacos , Oxigênio/metabolismo , Fosfatidilcolinas/metabolismo , RNA Mensageiro/metabolismo , Carneiro Doméstico , Volume de Ventilação PulmonarRESUMO
BACKGROUND: Exposure to high levels of oxygen (hyperoxia) after birth leads to lung injury. Our aims were to investigate the modulation of myeloid cell sub-populations and the reduction of fibrosis in the lungs following administration of human mesenchymal stem cells (hMSC) to neonatal mice exposed to hyperoxia. METHOD: Newborn mice were exposed to 90% O2 (hyperoxia) or 21% O2 (normoxia) from postnatal days 0-4. A sub-group of hyperoxia mice were injected intratracheally with 2.5X105 hMSCs. Using flow cytometry we assessed pulmonary immune cells at postnatal days 0, 4, 7 and 14. The following markers were chosen to identify these cells: CD45+ (leukocytes), Ly6C+Ly6G+ (granulocytes), CD11b+CD11c+ (macrophages); macrophage polarisation was assessed by F4/80 and CD206 expression. hMSCs expressing enhanced green fluorescent protein (eGFP) and firefly luciferase (fluc) were administered via the trachea at day 4. Lung macrophages in all groups were profiled using next generation sequencing (NGS) to assess alterations in macrophage phenotype. Pulmonary collagen deposition and morphometry were assessed at days 14 and 56 respectively. RESULTS: At day 4, hyperoxia increased the number of pulmonary Ly6C+Ly6G+ granulocytes and F4/80lowCD206low macrophages but decreased F4/80highCD206high macrophages. At days 7 and 14, hyperoxia increased numbers of CD45+ leukocytes, CD11b+CD11c+ alveolar macrophages and F4/80lowCD206low macrophages but decreased F4/80highCD206high macrophages. hMSCs administration ameliorated these effects of hyperoxia, notably reducing numbers of CD11b+CD11c+ and F4/80lowCD206low macrophages; in contrast, F4/80highCD206high macrophages were increased. Genes characteristic of anti-inflammatory 'M2' macrophages (Arg1, Stat6, Retnla, Mrc1, Il27ra, Chil3, and Il12b) were up-regulated, and pro-inflammatory 'M1' macrophages (Cd86, Stat1, Socs3, Slamf1, Tnf, Fcgr1, Il12b, Il6, Il1b, and Il27ra) were downregulated in isolated lung macrophages from hyperoxia-exposed mice administered hMSCs, compared to mice without hMSCs. Hydroxyproline assay at day 14 showed that the 2-fold increase in lung collagen following hyperoxia was reduced to control levels in mice administered hMSCs. By day 56 (early adulthood), hMSC administration had attenuated structural changes in hyperoxia-exposed lungs. CONCLUSIONS: Our findings suggest that hMSCs reduce neonatal lung injury caused by hyperoxia by modulation of macrophage phenotype. Not only did our cell-based therapy using hMSC induce structural repair, it limited the progression of pulmonary fibrosis.
Assuntos
Hiperóxia/metabolismo , Hiperóxia/terapia , Lesão Pulmonar/metabolismo , Lesão Pulmonar/terapia , Macrófagos Alveolares/metabolismo , Transplante de Células-Tronco Mesenquimais/métodos , Células Mieloides/metabolismo , Animais , Animais Recém-Nascidos , Feminino , Hiperóxia/patologia , Pulmão/metabolismo , Pulmão/patologia , Lesão Pulmonar/patologia , Macrófagos Alveolares/patologia , Células-Tronco Mesenquimais/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Células Mieloides/patologia , Gravidez , Resultado do TratamentoRESUMO
BackgroundAmong preterm infants, males have a greater incidence of respiratory distress and death than do females born at the same gestational age, likely due to sex-related differences in lung maturation. Our aim was to determine whether surfactant phospholipid composition differs between male and female preterm infants.MethodsGastric aspirate samples from male and female infants born between 25 and 30 weeks of gestation at The Royal Women's Hospital, Melbourne, Australia, were collected within 1 h after birth. Phospholipid composition was analyzed by electrospray ionization tandem mass spectrometry.ResultsPreterm males had higher proportions of total phosphatidylinositol (PI) and phosphatidylserine 36:2, lower proportions of total sphingomyelin (S) and S 33:1 and 35:1, and a greater phosphatidylcholine (PC)/S ratio than did females. The proportions of PC 30:0, PC 34:0, PC 34:2, PC 36:2, PC 36:3, and PC 38:2 differed between the sexes at different gestational weeks of birth; the proportion of PC 32:0 (dipalmitoylphosphatidylcholine) in males was lower than that in females at 25 weeks of gestation but higher at 27 weeks.ConclusionPhospholipid composition in pulmonary surfactant is different between male and female preterm infants of the same gestational age, which may contribute to the increased risk for respiratory morbidities in one sex.
Assuntos
Suco Gástrico/química , Lactente Extremamente Prematuro , Fosfolipídeos/análise , Surfactantes Pulmonares/química , Tensoativos/análise , Feminino , Idade Gestacional , Humanos , Recém-Nascido , Masculino , Fatores Sexuais , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem , VitóriaRESUMO
Supplemental oxygen (O2) increases the risk of lung injury in preterm infants, owing to an immature antioxidant system. Our objective was to determine whether impairing antioxidant defense by decreasing glutathione peroxidase 1 (GPx1) gene expression increases the injurious effects of hyperoxia (Hyp). GPx1+/+ and GPx1-/- C57Bl/6J mice were exposed to 21% O2 (Air) or 40% O2 (Hyp) from birth to postnatal day 7 (P7d); they were euthanized on P7d or maintained in air until adulthood [postnatal day 56 (P56d)] to assess short-term and long-term effects, respectively. We assessed lung architecture, three markers of pulmonary oxidative stress (P7d, P56d), macrophages in lung tissue (P7d), immune cells in bronchoalveolar lavage fluid (BALF; P56d), and GPx1-4 and catalase gene expression in lung tissue (P7d, P56d). On P7d, macrophages were decreased by lack of GPx1 expression and further decreased by hyperoxia. GPx1 expression was increased in GPx1+/+Hyp mice and decreased in both GPx1-/- groups. On P56d, heme oxygenase-1 was increased by hyperoxia when GPx1 was absent. There were significantly more immune cells from Hyp groups than from the GPx1+/+Air group and a greater proportion of lymphocytes in GPx1-/-Hyp mice. GPx1 expression was significantly decreased in GPx1-/- mice; GPx2-4 and catalase expression was increased in GPx1-/-Hyp mice compared with other groups. Tissue fraction was decreased in GPx1-/-Air mice; bronchiolar smooth muscle was decreased in GPx1-/- mice. GPx1 does not clearly exacerbate hyperoxia-induced increases in oxidative stress or lung injury but may alter pulmonary immune function. Increased expression of GPx2-4 and catalase in GPx1-/-Hyp mice suggests gene redundancy within the model.
Assuntos
Progressão da Doença , Regulação Enzimológica da Expressão Gênica , Glutationa Peroxidase/genética , Hiperóxia/enzimologia , Hiperóxia/genética , Lesão Pulmonar/enzimologia , Lesão Pulmonar/genética , Aldeídos/metabolismo , Animais , Animais Recém-Nascidos , Antioxidantes/metabolismo , Feminino , Glutationa Peroxidase/metabolismo , Heme Oxigenase-1/metabolismo , Pulmão/imunologia , Pulmão/patologia , Lesão Pulmonar/imunologia , Lesão Pulmonar/patologia , Masculino , Camundongos Endogâmicos C57BL , Estresse Oxidativo , Tirosina/análogos & derivados , Tirosina/metabolismo , Glutationa Peroxidase GPX1RESUMO
Purpose: To describe a mouse model of hyperoxia-induced vitreoretinopathy that replicated some of the clinical and pathologic features encountered in infants with severe retinopathy of prematurity and congenital ocular conditions such as persistent hyperplastic primary vitreous. Methods: Experimental mice (C57BL/6J) were exposed to 65% oxygen between postnatal days (P)0 to P7 and studied at P10, P14, and 3, 5, 8, 20, and 40 weeks. Controls were exposed to normoxic conditions. Fundus imaging and fluorescein angiography were performed at all time points, and spectral-domain optical coherence tomography (SD-OCT) and electroretinography were performed at 8- and 20-week time points. Eyes were processed for resin histology, frozen sections, and retinal whole mounts. Immunostaining was performed to visualize vasculature isolectin B4 (Ib4), collagen type IV, glial fibrillary acidic protein, and α-smooth muscle actin. Results: Early exposure to hyperoxia resulted in bilateral vitreous hemorrhages at 3 weeks. From 5 weeks onward there were extensive zones of retinal degeneration, scarring or gliosis, retinal folding, and detachments caused by traction of α-smooth muscle actin-positive vitreous membranes. Tortuous retinal vessels, together with hyperplastic and persistence of hyaloid vessels are evident into adulthood. In the early stages (P10-3 weeks), branches from the tunica vasculosa lentis (TVL) supplied the marginal retina until retinal vessels were established. The peripheral retina remained poorly vascularized into adulthood. Electroretinography revealed 50% to 60% diminution in retinal function in adult mice that strongly correlated with vitreal changes identified using SD-OCT. Conclusions: This animal model displays a mixture of vitreoretinal pathologic changes that persist into adulthood. The model may prove valuable in experimental investigations of therapeutic approaches to blinding conditions caused by vitreous and retinal abnormalities.
Assuntos
Hiperóxia/complicações , Oxigênio/metabolismo , Vasos Retinianos/patologia , Retinopatia da Prematuridade/etiologia , Tomografia de Coerência Óptica/métodos , Animais , Animais Recém-Nascidos , Modelos Animais de Doenças , Eletrorretinografia , Feminino , Angiofluoresceinografia , Seguimentos , Fundo de Olho , Hiperóxia/diagnóstico , Hiperóxia/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Vasos Retinianos/fisiopatologia , Retinopatia da Prematuridade/metabolismo , Retinopatia da Prematuridade/patologia , Retinopatia da Prematuridade/fisiopatologia , Índice de Gravidade de Doença , Fatores de TempoRESUMO
Many preterm infants require hyperoxic gas for survival, although it can contribute to lung injury. Experimentally, neonatal hyperoxia leads to persistent alterations in lung structure and increases leukocytes in bronchoalveolar lavage fluid (BALF). These effects of hyperoxia on the lungs are considered to be caused, at least in part, by increased oxidative stress. Our objective was to determine if dietary supplementation with a known source of antioxidants (tomato juice, TJ) could protect the developing lung from injury caused by breathing hyperoxic gas. Neonatal mice (C57BL6/J) breathed either 65% O2 (hyperoxia) or room air from birth until postnatal day 7 (P7d); some underwent necropsy at P7d and others were raised in room air until adulthood (P56d). In subsets of both groups, drinking water was replaced with TJ (diluted 50:50 in water) from late gestation to necropsy. At P7d and P56d, we analyzed total antioxidant capacity (TAC), markers of oxidative stress (nitrotyrosine and heme oxygenase-1 expression), inflammation (interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) expression), collagen (COL) and smooth muscle in the lungs; we also assessed lung structure. We quantified macrophages in lung tissue (at P7d) and leukocytes in BALF (at P56d). At P7d, TJ increased pulmonary TAC and COL1α1 expression and attenuated the hyperoxia-induced increase in nitrotyrosine and macrophage influx; however, changes in lung structure were not affected. At P56d, TJ increased TAC, decreased oxidative stress and reversed the hyperoxia-induced increase in bronchiolar smooth muscle. Additionally, TJ alone decreased IL-1ß expression, but following hyperoxia TJ increased TNF-α expression and did not alter the hyperoxia-induced increase in leukocyte number. We conclude that TJ supplementation during and after neonatal exposure to hyperoxia protects the lung from some but not all aspects of hyperoxia-induced injury, but may also have adverse side-effects. The effects of TJ are likely due to elevation of circulating antioxidant concentrations.
Assuntos
Lesão Pulmonar Aguda/dietoterapia , Sucos de Frutas e Vegetais , Hiperóxia/dietoterapia , Inflamação/dietoterapia , Solanum lycopersicum/química , Lesão Pulmonar Aguda/fisiopatologia , Animais , Animais Recém-Nascidos , Líquido da Lavagem Broncoalveolar , Dieta , Modelos Animais de Doenças , Hiperóxia/fisiopatologia , Inflamação/fisiopatologia , Pulmão/efeitos dos fármacos , Pulmão/fisiopatologia , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Alvéolos Pulmonares/efeitos dos fármacos , Alvéolos Pulmonares/fisiopatologiaRESUMO
BACKGROUND: Male preterm infants are more likely to experience respiratory distress syndrome than females. Our objectives were to determine if sex-related differences in physiological adaptation after preterm birth increase with time after birth and if the use of continuous positive airway pressure (CPAP) reduces these differences. METHODS: Unanesthetized lambs (9F, 8M) were delivered at 0.90 of term. Blood gases, metabolites, and cardiovascular and respiratory parameters were monitored in spontaneously breathing lambs for 8 h. Supplemental oxygen was administered via a face mask at 4 cmH2O CPAP. At 8 h, lung compliance was determined, and bronchoalveolar lavage fluid (BALF) was analyzed for total protein and surfactant phospholipids. Surfactant protein (SP) gene expression and protein expression of SP-A and pro-SP-C were determined in lung tissue. RESULTS: For 8 h after delivery, males had significantly lower arterial pH and higher Paco2, and a greater percentage of males were dependent on supplemental oxygen than females. Inspiratory effort was greater and lung compliance was lower in male lambs. Total protein concentration in BALF, SP gene expression, and SP-A protein levels were not different between sexes; pro-SP-C was 24% lower in males. CONCLUSION: The use of CPAP did not eliminate the male disadvantage, which continues for up to 8 h after preterm birth.
Assuntos
Nascimento Prematuro/fisiopatologia , Proteína A Associada a Surfactante Pulmonar/química , Proteína B Associada a Surfactante Pulmonar/química , Síndrome do Desconforto Respiratório do Recém-Nascido/fisiopatologia , Sistema Respiratório/fisiopatologia , Caracteres Sexuais , Adaptação Fisiológica , Animais , Animais Recém-Nascidos , Peso Corporal , Líquido da Lavagem Broncoalveolar/química , Dióxido de Carbono/sangue , Pressão Positiva Contínua nas Vias Aéreas , Feminino , Maturidade dos Órgãos Fetais , Regulação da Expressão Gênica no Desenvolvimento , Concentração de Íons de Hidrogênio , Pulmão/embriologia , Pulmão/metabolismo , Complacência Pulmonar , Masculino , Tamanho do Órgão , Oxigênio/administração & dosagem , Oxigênio/sangue , Fosfolipídeos/análise , Nascimento Prematuro/metabolismo , Proteína A Associada a Surfactante Pulmonar/biossíntese , Proteína A Associada a Surfactante Pulmonar/genética , Proteína B Associada a Surfactante Pulmonar/biossíntese , Proteína B Associada a Surfactante Pulmonar/genética , Síndrome do Desconforto Respiratório do Recém-Nascido/sangue , Ovinos , Vísceras/anatomia & histologiaRESUMO
Intrauterine growth restriction (IUGR) and preterm birth are frequent comorbidities and, combined, increase the risk of adverse respiratory outcomes compared with that in appropriately grown (AG) infants. Potential underlying reasons for this increased respiratory morbidity in IUGR infants compared with AG infants include altered fetal lung development, fetal lung inflammation, increased respiratory requirements, and/or increased ventilation-induced lung injury. IUGR was surgically induced in preterm fetal sheep (0.7 gestation) by ligation of a single umbilical artery. Four weeks later, preterm lambs were euthanized at delivery or delivered and ventilated for 2 h before euthanasia. Ventilator requirements, lung inflammation, early markers of lung injury, and morphological changes in lung parenchymal and vascular structure and surfactant composition were analyzed. IUGR preterm lambs weighed 30% less than AG preterm lambs, with increased brain-to-body weight ratio, indicating brain sparing. IUGR did not induce lung inflammation or injury or alter lung parenchymal and vascular structure compared with AG fetuses. IUGR and AG lambs had similar oxygenation and respiratory requirements after birth and had significant, but similar, increases in proinflammatory cytokine expression, lung injury markers, gene expression, and surfactant phosphatidylcholine species compared with unventilated controls. IUGR does not induce pulmonary structural changes in our model. Furthermore, IUGR and AG preterm lambs have similar ventilator requirements in the immediate postnatal period. This study suggests that increased morbidity and mortality in IUGR infants is not due to altered lung tissue or vascular structure, or to an altered response to early ventilation.
Assuntos
Retardo do Crescimento Fetal/metabolismo , Pulmão/metabolismo , Pneumonia/metabolismo , Surfactantes Pulmonares/metabolismo , Lesão Pulmonar Induzida por Ventilação Mecânica/metabolismo , Animais , Animais Recém-Nascidos , Feminino , Idade Gestacional , Gravidez , Respiração Artificial/efeitos adversos , OvinosRESUMO
Infants born very preterm are usually exposed to high oxygen concentrations but this may impair lung function in survivors in later life. However, the precise changes involved are poorly understood. We determined how neonatal hyperoxia alters lung function at mid-adulthood in mice. Neonatal C57BL/6J mice inhaled 65% oxygen (HE group) from birth for 7 days. They then breathed room air until 11 months of age (P11mo); these mice experienced growth restriction. Controls breathed only room air. To exclude the effects of growth restriction, a group of dams was rotated between hyperoxia and normoxia during the exposure period (HE+DR group). Lung function was measured at P11mo. HE mice had increased inspiratory capacity, work of breathing and tissue damping. HE+DR mice had further increases in inspiratory capacity and work of breathing, and reduced FEV100/FVC. Total lung capacity was increased in HE+DR males. HE males had elevated responses to methacholine. Neonatal hyperoxia alters lung function at mid-adulthood, especially in males.
Assuntos
Hiperóxia/fisiopatologia , Pulmão/crescimento & desenvolvimento , Pulmão/fisiopatologia , Animais , Animais Recém-Nascidos , Peso Corporal , Broncoconstritores/farmacologia , Modelos Animais de Doenças , Feminino , Pulmão/efeitos dos fármacos , Medidas de Volume Pulmonar , Masculino , Cloreto de Metacolina/farmacologia , Camundongos Endogâmicos C57BL , Caracteres SexuaisRESUMO
Preterm infants often require supplemental oxygen due to lung immaturity, but hyperoxia can contribute to an increased risk of respiratory illness later in life. Our aim was to compare the effects of mild and moderate levels of neonatal hyperoxia on markers of pulmonary oxidative stress and inflammation and on lung architecture; both immediate and persistent effects were assessed. Neonatal mice (C57BL6/J) were raised in either room air (21% O2), mild (40% O2), or moderate (65% O2) hyperoxia from birth until postnatal day 7 (P7d). The mice were killed at either P7d (immediate effects) or lived in air until adulthood (P56d, persistent effects). We enumerated macrophages in lung tissue at P7d and immune cells in bronchoalveolar lavage fluid (BALF) at P56d. At P7d and P56d, we assessed pulmonary oxidative stress [heme oxygenase-1 (HO-1) and nitrotyrosine staining] and lung architecture. The data were interrogated for sex differences. At P7d, HO-1 gene expression was greater in the 65% O2 group than in the 21% O2 group. At P56d, the area of nitrotyrosine staining and number of immune cells were greater in the 40% O2 and 65% O2 groups relative to the 21% O2 group. Exposure to 65% O2, but not 40% O2, led to larger alveoli and lower tissue fraction in the short term and to persistently fewer bronchiolar-alveolar attachments. Exposure to 40% O2 or 65% O2 causes persistent increases in pulmonary oxidative stress and immune cells, suggesting chronic inflammation within the adult lung. Unlike 65% O2, 40% O2 does not affect lung architecture.
Assuntos
Hiperóxia/fisiopatologia , Macrófagos Alveolares/citologia , Estresse Oxidativo/fisiologia , Oxigênio/efeitos adversos , Alvéolos Pulmonares/fisiopatologia , Animais , Animais Recém-Nascidos , Líquido da Lavagem Broncoalveolar/citologia , Feminino , Heme Oxigenase-1/biossíntese , Inflamação/patologia , Proteínas de Membrana/biossíntese , Camundongos , Camundongos Endogâmicos C57BL , Estresse Oxidativo/efeitos dos fármacos , Oxigênio/farmacologia , Alvéolos Pulmonares/metabolismo , Tirosina/análogos & derivadosRESUMO
BACKGROUND: Lung immaturity due to preterm birth is a significant complication affecting neonatal health. Despite the detrimental effects of supplemental oxygen on alveolar formation, it remains an important treatment for infants with respiratory distress. Macrophages are traditionally associated with the propagation of inflammatory insults, however increased appreciation of their diversity has revealed essential functions in development and regeneration. METHODS: Macrophage regulatory cytokine Colony-Stimulating Factor-1 (CSF-1) was investigated in a model of neonatal hyperoxia exposure, with the aim of promoting macrophages associated with alveologenesis to protect/rescue lung development and function. Neonatal mice were exposed to normoxia (21% oxygen) or hyperoxia (Hyp; 65% oxygen); and administered CSF-1 (0.5 µg/g, daily × 5) or vehicle (PBS) in two treatment regimes; 1) after hyperoxia from postnatal day (P)7-11, or 2) concurrently with five days of hyperoxia from P1-5. Lung structure, function and macrophages were assessed using alveolar morphometry, barometric whole-body plethysmography and flow cytometry. RESULTS AND DISCUSSION: Seven days of hyperoxia resulted in an 18% decrease in body weight and perturbation of lung structure and function. In regime 1, growth restriction persisted in the Hyp + PBS and Hyp + CSF-1 groups, although perturbations in respiratory function were resolved by P35. CSF-1 increased CSF-1R+/F4/80+ macrophage number by 34% at P11 compared to Hyp + PBS, but was not associated with growth or lung structural rescue. In regime 2, five days of hyperoxia did not cause initial growth restriction in the Hyp + PBS and Hyp + CSF-1 groups, although body weight was decreased at P35 with CSF-1. CSF-1 was not associated with increased macrophages, or with functional perturbation in the adult. Overall, CSF-1 did not rescue the growth and lung defects associated with hyperoxia in this model; however, an increase in CSF-1R+ macrophages was not associated with an exacerbation of lung injury. The trophic functions of macrophages in lung development requires further elucidation in order to explore macrophage modulation as a strategy for promoting lung maturation.
Assuntos
Hiperóxia/tratamento farmacológico , Lesão Pulmonar/tratamento farmacológico , Pulmão/efeitos dos fármacos , Fator Estimulador de Colônias de Macrófagos/administração & dosagem , Macrófagos Alveolares/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Peso Corporal , Modelos Animais de Doenças , Esquema de Medicação , Genes Reporter , Proteínas de Fluorescência Verde/biossíntese , Proteínas de Fluorescência Verde/genética , Hiperóxia/imunologia , Hiperóxia/fisiopatologia , Pulmão/crescimento & desenvolvimento , Pulmão/imunologia , Pulmão/fisiopatologia , Lesão Pulmonar/imunologia , Lesão Pulmonar/fisiopatologia , Macrófagos Alveolares/imunologia , Camundongos Transgênicos , Regiões Promotoras Genéticas , Receptor de Fator Estimulador de Colônias de Macrófagos/genética , Respiração , Testes de Função Respiratória , Fatores de TempoRESUMO
While the impact of alcohol consumption by pregnant women on fetal neurodevelopment has received much attention, the effects on the cardiovascular system are not well understood. We hypothesised that repeated exposure to alcohol (ethanol) in utero would alter fetal arterial reactivity and wall stiffness, key mechanisms leading to cardiovascular disease in adulthood. Ethanol (0.75 g (kg body weight)(-1)) was infused intravenously into ewes over 1 h daily for 39 days in late pregnancy (days 95-133 of pregnancy, term â¼147 days). Maternal and fetal plasma ethanol concentrations at the end of the hour were â¼115 mg dl(-1), and then declined to apparent zero over 8 h. At necropsy (day 134), fetal body weight and fetal brain-body weight ratio were not affected by alcohol infusion. Small arteries (250-300 µm outside diameter) from coronary, renal, mesenteric, femoral (psoas) and cerebral beds were isolated. Endothelium-dependent vasodilatation sensitivity was reduced 10-fold in coronary resistance arteries, associated with a reduction in endothelial nitric oxide synthase mRNA (P = 0.008). Conversely, vasodilatation sensitivity was enhanced 10-fold in mesenteric and renal resistance arteries. Arterial stiffness was markedly increased (P = 0.0001) in all five vascular beds associated with an increase in elastic modulus and, in cerebral vessels, with an increase in collagen Iα mRNA. Thus, we show for the first time that fetal arteries undergo marked and regionally variable adaptations as a consequence of repeated alcohol exposure. These alcohol-induced vascular effects occurred in the apparent absence of fetal physical abnormalities or fetal growth restriction.
Assuntos
Consumo de Bebidas Alcoólicas/efeitos adversos , Feto/efeitos dos fármacos , Troca Materno-Fetal , Rigidez Vascular/efeitos dos fármacos , Vasodilatação/efeitos dos fármacos , Animais , Artérias/efeitos dos fármacos , Artérias/fisiologia , Encéfalo/irrigação sanguínea , Encéfalo/fisiologia , Vasos Coronários/efeitos dos fármacos , Vasos Coronários/fisiologia , Feminino , Feto/fisiologia , Trato Gastrointestinal/irrigação sanguínea , Trato Gastrointestinal/fisiologia , Rim/irrigação sanguínea , Rim/fisiologia , Músculo Esquelético/irrigação sanguínea , Músculo Esquelético/fisiologia , Gravidez , Ovinos , Vasodilatação/fisiologiaRESUMO
Preterm infants who receive supplemental oxygen for prolonged periods are at increased risk of impaired lung function later in life. This suggests that neonatal hyperoxia induces persistent changes in small conducting airways (bronchioles). Although the effects of neonatal hyperoxia on alveolarization are well documented, little is known about its effects on developing bronchioles. We hypothesized that neonatal hyperoxia would remodel the bronchiolar walls, contributing to altered lung function in adulthood. We studied three groups of mice (C57BL/6J) to postnatal day 56 (P56; adulthood) when they either underwent lung function testing or necropsy for histological analysis of the bronchiolar wall. One group inhaled 65% O2 from birth until P7, after which they breathed room air; this group experienced growth restriction (HE+GR group). We also used a group in which hyperoxia-induced GR was prevented by dam rotation (HE group). A control group inhaled room air from birth. At P56, the bronchiolar epithelium of HE mice contained fewer Clara cells and more ciliated cells, and the bronchiolar wall contained â¼25% less collagen than controls; in HE+GR mice the bronchiolar walls had â¼13% more collagen than controls. Male HE and HE+GR mice had significantly thicker bronchiolar epithelium than control males and altered lung function (HE males: greater dynamic compliance; HE+GR males: lower dynamic compliance). We conclude that neonatal hyperoxia remodels the bronchiolar wall and, in adult males, affects lung function, but effects are altered by concomitant growth restriction. Our findings may partly explain the reports of poor lung function in ex-preterm children and adults.
Assuntos
Bronquíolos/citologia , Bronquíolos/fisiologia , Hiperóxia/fisiopatologia , Alvéolos Pulmonares/fisiologia , Animais , Animais Recém-Nascidos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Alvéolos Pulmonares/citologia , Testes de Função RespiratóriaRESUMO
Preterm male infants have a higher incidence of morbidity and mortality due to respiratory insufficiency than females of the same gestational age. This male disadvantage could be due to differences in lung architecture; however, few studies have compared lung architecture in male and female fetuses during late gestation. Our principal objectives were to compare the morphology of the fetal lung and the maturity of the surfactant system in preterm male and female fetuses. Lungs from male (n = 9) and female (n = 11) fetal sheep were collected at 0.9 of term (131 days of the 145-day gestation) for morphological and molecular analyses. In separate groups, tracheal liquid was obtained from male (n = 9) and female (n = 9) fetuses at 0.9 of term for determination of surfactant phospholipid composition. We found no sex-related differences in body weight, lung weight, right lung volume, lung tissue and airspace fractions, mean linear intercept, septal crest density, septal thickness, the proportion of proliferating and apoptotic cells, and the percentages of collagen or elastin. The gene expression of surfactant protein -A, -B, -C, and -D and tropoelastin was similar between sexes. There were no differences in the proportion of the major phospholipid classes in the tracheal liquid between sexes; however there was a significantly higher percentage of the phospholipid species phosphatidylinositol 38:5 in males. The greater morbidity and mortality in preterm male lambs do not appear to be related to differences in lung structure or surfactant phospholipid synthesis before birth, but may relate to physiological adaptation to air-breathing at birth.
Assuntos
Animais Recém-Nascidos/fisiologia , Maturidade dos Órgãos Fetais/fisiologia , Feto/fisiologia , Pulmão/fisiologia , Traqueia/fisiologia , Animais , Animais Recém-Nascidos/metabolismo , Peso Corporal/fisiologia , Feminino , Feto/metabolismo , Pulmão/metabolismo , Masculino , Fosfolipídeos/metabolismo , Surfactantes Pulmonares/metabolismo , Respiração , Ovinos/metabolismo , Ovinos/fisiologia , Traqueia/metabolismoRESUMO
BACKGROUND: Supplemental oxygen is necessary in the respiratory support of very preterm infants, but it may contribute to bronchopulmonary dysplasia and an increased risk of poor lung function in later life. It is well established that hyperoxia can inhibit alveolarization, but effects on the developing conducting airways, which are important determinants of lung function, are poorly understood. It is possible that prolonged exposure of the immature lung to hyperoxic gas alters the development of small conducting airways (bronchioles), and that these effects may persist throughout life. OBJECTIVES: To examine the effects of neonatal inhalation of hyperoxic gas on the bronchiolar walls in adulthood. METHODS: Neonatal mice (C57BL/6J) born at term inhaled 65% O2 from birth until postnatal day 7; thereafter, they were raised in room air until 10 months postnatal age (P10mo), which is advanced adulthood. Age-matched controls inhaled room air from birth. We investigated small conducting airways with a diameter between 105-310 µm. RESULTS: At P10mo, bronchiolar walls of hyperoxia-exposed mice contained â¼18% more smooth muscle than controls (p < 0.05), although there was no effect on bronchiolar epithelium or collagen. Neonatal hyperoxia resulted in significantly fewer bronchiolar-alveolar attachments at P10mo (p < 0.05); this was accompanied by persistent simplification of the lung parenchyma, as indicated by greater mean linear intercept and less parenchymal tissue (p < 0.05). CONCLUSIONS: Neonatal exposure to hyperoxia induces remodeling of the bronchiolar walls and loss of bronchiolar-alveolar attachments in adulthood, both of which could contribute to impaired lung function and airway hyper-reactivity.
Assuntos
Envelhecimento/fisiologia , Remodelação das Vias Aéreas/fisiologia , Animais Recém-Nascidos/fisiologia , Bronquíolos/fisiopatologia , Hiperóxia/complicações , Hiperóxia/fisiopatologia , Envelhecimento/patologia , Remodelação das Vias Aéreas/efeitos dos fármacos , Animais , Hiper-Reatividade Brônquica/epidemiologia , Hiper-Reatividade Brônquica/fisiopatologia , Bronquíolos/efeitos dos fármacos , Bronquíolos/patologia , Displasia Broncopulmonar/epidemiologia , Displasia Broncopulmonar/fisiopatologia , Modelos Animais de Doenças , Feminino , Pulmão/efeitos dos fármacos , Pulmão/patologia , Pulmão/fisiopatologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Liso/efeitos dos fármacos , Músculo Liso/patologia , Músculo Liso/fisiopatologia , Oxigênio/farmacologia , Alvéolos Pulmonares/efeitos dos fármacos , Alvéolos Pulmonares/patologia , Alvéolos Pulmonares/fisiopatologia , Fatores de RiscoRESUMO
High levels of alcohol (ethanol) exposure during fetal life can affect liver development and can increase susceptibility to infection after birth. Our aim was to determine the effects of a moderate level of ethanol exposure in late gestation on the morphology, iron status, and inflammatory status of the ovine fetal liver. Pregnant ewes were chronically catheterized at 91 days of gestation (DG; term ~145 DG) for daily intravenous infusion of ethanol (0.75 g/kg maternal body wt; n = 8) or saline (n = 7) over 1 h from 95 to 133 DG. At necropsy (134 DG), fetal livers were collected for analysis. Liver weight, general liver morphology, hepatic cell proliferation and apoptosis, perivascular collagen deposition, and interleukin (IL)-1ß, IL-6, or IL-8 mRNA levels were not different between groups. However, ethanol exposure led to significant decreases in hepatic content of ferric iron and gene expression of the iron-regulating hormone hepcidin and tumor necrosis factor (TNF)-α (all P < 0.05). In the placenta, there was no difference in transferrin receptor, divalent metal transporter 1, and ferritin mRNA levels; however, ferroportin mRNA levels were increased in ethanol-exposed animals (P < 0.05), and ferroportin protein tended to be increased (P = 0.054). Plasma iron concentration was not different between control and ethanol-exposed groups; control fetuses had significantly higher iron concentrations than their mothers, whereas maternal and fetal iron concentrations were similar in ethanol-exposed animals. We conclude that daily ethanol exposure during the third-trimester-equivalent in sheep does not alter fetal liver morphology; however, decreased fetal liver ferric iron content and altered hepcidin and ferroportin gene expression indicate that iron homeostasis is altered.
Assuntos
Etanol/efeitos adversos , Feto/metabolismo , Homeostase/fisiologia , Ferro/metabolismo , Fígado/metabolismo , Fígado/patologia , Prenhez/metabolismo , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Animais , Peptídeos Catiônicos Antimicrobianos/metabolismo , Apoptose/efeitos dos fármacos , Proteínas de Transporte de Cátions/metabolismo , Relação Dose-Resposta a Droga , Etanol/farmacologia , Feminino , Desenvolvimento Fetal/efeitos dos fármacos , Hepcidinas , Homeostase/efeitos dos fármacos , Fígado/efeitos dos fármacos , Modelos Animais , Tamanho do Órgão/efeitos dos fármacos , Placenta/metabolismo , Gravidez , OvinosRESUMO
Preterm birth affects 8-10% of human pregnancies and is a major cause of long-term disability. Individuals who are born very preterm, especially if they develop bronchopulmonary dysplasia (BPD), have an increased risk of impaired lung function in infancy, childhood and adulthood, as well as an increased risk of respiratory illness. Our aim is to briefly review current understanding of the basis for long-term impairments in lung function and respiratory health following preterm birth and BPD. Histopathology of the lungs of infants and children following preterm birth and BPD shows altered development of the lung parenchyma, conducting airways and pulmonary vasculature. Owing to improvements in the care of preterm infants, especially the use of exogenous surfactant and lower concentrations of administered oxygen, lung pathology following preterm birth and BPD is less severe than in the past. Recent studies indicate that very preterm birth and BPD can lead to hyperplasia of airway smooth muscle, impaired alveolarization, pulmonary inflammation and an increase in pulmonary artery muscularization. Imaging of adult lungs suggests that the deficit in alveoli can persist into later life. Long-term lung injury apparently relates to the use of mechanical ventilation and the use of supplemental oxygen in infancy. Impaired lung function in later life is due to airway hyper-reactivity and fewer alveoli, resulting in reductions in the surface area for gas exchange and physical support for bronchioles. Because the incidence of preterm birth is not declining, it will continue to be a major cause of respiratory ill-health in adults.
Assuntos
Desenvolvimento do Adolescente , Envelhecimento , Displasia Broncopulmonar/fisiopatologia , Desenvolvimento Infantil , Pulmão/fisiopatologia , Nascimento Prematuro/fisiopatologia , Adolescente , Adulto , Displasia Broncopulmonar/patologia , Displasia Broncopulmonar/terapia , Criança , Tolerância ao Exercício , Humanos , Hipertensão Pulmonar/etiologia , Hipertensão Pulmonar/fisiopatologia , Hipertensão Pulmonar/prevenção & controle , Recém-Nascido , Pulmão/irrigação sanguínea , Pulmão/patologia , Nascimento Prematuro/epidemiologia , Nascimento Prematuro/patologia , Prevalência , Surfactantes Pulmonares/uso terapêutico , Índice de Gravidade de DoençaRESUMO
Preterm neonates are born while nephrogenesis is ongoing and are commonly exposed to factors in the extrauterine environment that may impair renal development. Supplemental oxygen therapy exposes the preterm infant to a hyperoxic environment that may induce oxidative stress. Our aim was to determine the immediate and long-term effects of exposure to hyperoxia, during the period of postnatal nephrogenesis, on renal development. Newborn mice (C57BL/6J) were kept in a normoxic (room air, 21% oxygen) or a controlled hyperoxic (65% oxygen) environment from birth to postnatal day 7 (P7d). From P7d, animals were maintained in room air until early adulthood at postnatal day 56 (P56d) or middle age (10 mo; P10mo). Pups were assessed for glomerular maturity and renal corpuscle cross-sectional area at P7d (control n = 14; hyperoxic n = 14). Nephron number and renal corpuscle size were determined stereologically at P56d (control n = 14; hyperoxic n = 14) and P10mo (control n = 10; hyperoxic n = 10). At P7d, there was no effect of hyperoxia on glomerular size or maturity. In early adulthood (P56d), body weights, relative kidney weights and volumes, and nephron number were not different between groups, but the renal corpuscles were significantly enlarged. This was no longer evident at P10mo, with relative kidney weights and volumes, nephron number, and renal corpuscle size not different between groups. Furthermore, hyperoxia exposure did not significantly accelerate glomerulosclerosis in middle age. Hence, our findings show no overt long-term deleterious effects of early life hyperoxia on glomerular structure.
Assuntos
Hiperóxia/patologia , Nefropatias/patologia , Glomérulos Renais/crescimento & desenvolvimento , Rim/crescimento & desenvolvimento , Animais , Peso Corporal , Proliferação de Células , Feminino , Rim/patologia , Glomérulos Renais/patologia , Masculino , CamundongosRESUMO
Male preterm infants are at greater risk of respiratory morbidity and mortality than females but mechanisms are poorly understood. Our objective was to identify the basis for the "male disadvantage" following preterm birth using an ovine model of preterm birth in which survival of females is greater than males. At 0.85 of term, fetal sheep underwent surgery (11 female, 10 male) for the implantation of vascular catheters to monitor blood gases and arterial pressure. After cesarean delivery at 0.90 of term, lambs were monitored for 4 h while spontaneously breathing; lambs were then euthanized and static lung compliance measured. We analyzed surfactant phospholipid composition in amniotic fluid and in bronchoalveolar lavage fluid (BALF) taken at necropsy; we also analyzed surfactant protein (SP) expression in lung tissue. Before delivery male fetuses tended to have lower pH (P = 0.052) compared with females. One hour after delivery, males had significantly lower pH and higher arterial partial pressure of CO(2) (Pa(CO(2))), lactate, glucose, and mean arterial pressure than females. Two males died 1 h after birth. Static lung compliance was 37% lower in males than females (P < 0.05). In BALF, males had significantly more protein, a lower percentage of the phosphatidylcholine (PC) 32:0 (dipalmitoylphosphatidylcholine) and higher percentages of PC34:2 and PC36:2. There were no sex-related differences in lung architecture or expression of SP-A, -B, -C, and -D. The lower lung compliance in male preterm lambs compared with females may be due to altered surfactant phospholipid composition and function. These changes may compromise gas exchange and impair respiratory adaptation after male preterm birth.