RESUMEN
Optimal oxygen saturation as measured by pulse oximetry (SpO2) in neonatal lung injury, such as meconium aspiration syndrome (MAS) and persistent pulmonary hypertension of newborn (PPHN), is not known. Our goal was to determine the SpO2 range in lambs with MAS and PPHN that results in the highest brain oxygen delivery (bDO2) and pulmonary blood flow (Qp) and the lowest pulmonary vascular resistance and oxidative stress. Meconium was instilled into endotracheal tubes in 25 near-term gestation lambs, and the umbilical cord was occluded to induce asphyxia and gasping, causing MAS and PPHN. Lambs were randomized into four groups and ventilated for 6 hours with fixed fraction of inspired oxygen (FiO2) = 1.0 irrespective of SpO2, and three groups had FiO2 titrated to keep preductal SpO2 between 85% and 89%, 90% and 94%, and 95% and 99%, respectively. Tissues were collected to measure nitric oxide synthase activity, 3-nitrotyrosine, and 8-isoprostanes. Throughout the 6-hour exposure period, lambs in the 95-99% SpO2 target group had the highest Qp, lowest pulmonary vascular resistance, and highest bDO2 but were exposed to higher FiO2 (0.5 ± 0.21 vs. 0.29 ± 0.17) with higher lung 3-nitrotyrosine (0.67 [interquartile range (IQR), 0.43-0.73] ng/mcg protein vs. 0.1 [IQR, 0.09-0.2] ng/mcg protein) and lower lung nitric oxide synthase activity (196 [IQR, 192-201] mMol nitrite/mg protein vs. 270 [IQR, 227-280] mMol nitrite/mg protein) compared with the 90-94% target group. Brain 3-nitrotyrosine was lower in the 85-89% target group, and brain/lung 8-isoprostane levels were not significantly different. In term lambs with MAS and PPHN, Qp and bDO2 through the first 6 hours are higher with target SpO2 in the 95-99% range. However, the 90-94% target range is associated with significantly lower FiO2 and lung oxidative stress. Clinical trials comparing the 90-94% versus the 95-99% SpO2 target range in term infants with PPHN are warranted.
Asunto(s)
Hipertensión Pulmonar/metabolismo , Pulmón/metabolismo , Síndrome de Aspiración de Meconio/metabolismo , Oxígeno/metabolismo , Animales , Animales Recién Nacidos , Dinoprost/análogos & derivados , Dinoprost/farmacología , Femenino , Hipertensión Pulmonar/tratamiento farmacológico , Pulmón/efectos de los fármacos , Masculino , Síndrome de Aspiración de Meconio/tratamiento farmacológico , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa/metabolismo , Estrés Oxidativo/efectos de los fármacos , Oximetría/métodos , Síndrome de Circulación Fetal Persistente/tratamiento farmacológico , Síndrome de Circulación Fetal Persistente/metabolismo , Embarazo , Arteria Pulmonar/efectos de los fármacos , Arteria Pulmonar/metabolismo , Ovinos/metabolismo , Tirosina/análogos & derivados , Tirosina/farmacología , Resistencia Vascular/efectos de los fármacosRESUMEN
No in vivo data are available regarding the effect of meconium on human surfactant in the early stages of severe meconium aspiration syndrome (MAS). In the present study, we sought to characterize the changes in surfactant composition, function, and structure during the early phase of meconium injury. We designed a translational prospective cohort study of nonbronchoscopic BAL of neonates with severe MAS (n = 14) or no lung disease (n = 18). Surfactant lipids were analyzed by liquid chromatography-high-resolution mass spectrometry. Secretory phospholipase A2 subtypes IB, V, and X and SP-A (surfactant protein A) were assayed by ELISA. SP-B and SP-C were analyzed by Western blotting under both nonreducing and reducing conditions. Surfactant function was assessed by adsorption test and captive bubble surfactometry, and lung aeration was evaluated by semiquantitative lung ultrasound. Surfactant nanostructure was studied using cryo-EM and atomic force microscopy. Several changes in phospholipid subclasses were detected during MAS. Lysophosphatidylcholine species released by phospholipase A2 hydrolysis were increased. SP-B and SP-C were significantly increased together with some shorter immature forms of SP-B. Surfactant function was impaired and correlated with poor lung aeration. Surfactant nanostructure was significantly damaged in terms of vesicle size, tridimensional complexity, and compactness. Various alterations of surfactant phospholipids and proteins were detected in the early phase of severe meconium aspiration and were due to hydrolysis and inflammation and a defensive response. This impairs both surfactant structure and function, finally resulting in reduced lung aeration. These findings support the development of new surfactant protection and antiinflammatory strategies for severe MAS.
Asunto(s)
Pulmón/efectos de los fármacos , Síndrome de Aspiración de Meconio/tratamiento farmacológico , Surfactantes Pulmonares/farmacología , Tensoactivos/farmacología , Antiinflamatorios/farmacología , Humanos , Recién Nacido , Pulmón/metabolismo , Síndrome de Aspiración de Meconio/metabolismo , Síndrome de Aspiración de Meconio/fisiopatología , Fosfolipasas A2/efectos de los fármacos , Fosfolipasas A2/metabolismo , Fosfolípidos/metabolismo , Surfactantes Pulmonares/metabolismoRESUMEN
Acute respiratory distress syndrome (ARDS) is characterized by diffuse lung damage, inflammation, oedema formation, and surfactant dysfunction leading to hypoxemia. Severe ARDS can accelerate the injury of other organs, worsening the patient´s status. There is an evidence that the lung tissue injury affects the right heart function causing cor pulmonale. However, heart tissue changes associated with ARDS are still poorly known. Therefore, this study evaluated oxidative and inflammatory modifications of the heart tissue in two experimental models of ARDS induced in New Zealand rabbits by intratracheal instillation of neonatal meconium (100 mg/kg) or by repetitive lung lavages with saline (30 ml/kg). Since induction of the respiratory insufficiency, all animals were oxygen-ventilated for next 5 h. Total and differential counts of leukocytes were measured in the arterial blood, markers of myocardial injury [(troponin, creatine kinase - myocardial band (CK-MB), lactate dehydrogenase (LD)] in the plasma, and markers of inflammation [tumour necrosis factor (TNF)alpha, interleukin (IL)-6], cardiovascular risk [galectin-3 (Gal-3)], oxidative changes [thiobarbituric acid reactive substances (TBARS), 3-nitrotyrosine (3NT)], and vascular damage [receptor for advanced glycation end products (RAGE)] in the heart tissue. Apoptosis of heart cells was investigated immunohistochemically. In both ARDS models, counts of total leukocytes and neutrophils in the blood, markers of myocardial injury, inflammation, oxidative and vascular damage in the plasma and heart tissue, and heart cell apoptosis increased compared to controls. This study indicates that changes associated with ARDS may contribute to early heart damage what can potentially deteriorate the cardiac function and contribute to its failure.
Asunto(s)
Lesiones Cardíacas/patología , Inflamación/patología , Lesión Pulmonar/patología , Síndrome de Dificultad Respiratoria/patología , Animales , Apoptosis/fisiología , Biomarcadores/metabolismo , Modelos Animales de Enfermedad , Femenino , Lesiones Cardíacas/metabolismo , Inflamación/metabolismo , Lesión Pulmonar/metabolismo , Masculino , Síndrome de Aspiración de Meconio/metabolismo , Síndrome de Aspiración de Meconio/patología , Estrés Oxidativo/fisiología , Conejos , Síndrome de Dificultad Respiratoria/metabolismoRESUMEN
This study aimed to evaluate the molecular background of N-acetylcysteine (NAC) and recombinant human superoxide dismutase (rhSOD) antioxidant action when combined with exogenous surfactant in the treatment of meconium aspiration syndrome (MAS), considering redox signalling a principal part of cell response to meconium. Young New Zealand rabbits were instilled with meconium suspension (Mec) and treated by surfactant alone (Surf) or surfactant in combination with i.v. NAC (Surf + NAC) or i.t. rhSOD (Surf + SOD), and oxygen-ventilated for 5 h. Dynamic lung-thorax compliance, mean airway pressure, PaO2/FiO2 and ventilation efficiency index were evaluated every hour; post mortem, inflammatory and oxidative markers (advanced oxidation protein products, total antioxidant capacity, hydroxynonenal (HNE), p38 mitogen activated protein kinase, caspase 3, thromboxane, endothelin-1 and secretory phospholipase A2) were assessed in pulmonary tissue homogenates. rhSOD addition to surfactant improved significantly, but transiently, gas exchange and reduced levels of inflammatory and oxidative molecules with higher impact; Surf + NAC had stronger effect only on HNE formation, and duration of treatment efficacy in respiratory parameters. In both antioxidants, it seems that targeting reactive oxygen species may be strong supporting factor in surfactant treatment of MAS due to redox sensitivity of many intracellular pathways triggered by meconium.
Asunto(s)
Acetilcisteína/farmacología , Proteínas Recombinantes/farmacología , Superóxido Dismutasa/farmacología , Tensoactivos/farmacología , Animales , Apoptosis , Biomarcadores , Modelos Animales de Enfermedad , Humanos , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Pulmón/fisiopatología , Rendimiento Pulmonar/efectos de los fármacos , Síndrome de Aspiración de Meconio/tratamiento farmacológico , Síndrome de Aspiración de Meconio/etiología , Síndrome de Aspiración de Meconio/metabolismo , Síndrome de Aspiración de Meconio/fisiopatología , Oxidación-Reducción , Estrés Oxidativo/efectos de los fármacos , Conejos , Especies Reactivas de Oxígeno/metabolismo , Pruebas de Función RespiratoriaRESUMEN
PURPOSE: To evaluate the effects of hypothermia treatment on meconium-induced inflammation. METHODS: Fifteen rats were instilled with human meconium (MEC, 1.5 mL/kg, 65 mg/mL) intratracheally and ventilated for 3 hours. Eight rats that were ventilated and not instilled with meconium served as a sham group. In MEC-hypothermia group, the body temperature was lowered to 33±0.5°C. Analysis of the blood gases, interleukin (IL)-1ß, IL-6, IL-8, and tumor necrosis factor (TNF)-α in bronchoalveolar lavage (BAL) fluid samples, and histological analyses of the lungs were performed. RESULTS: The BAL fluid TNF-α, IL-1ß, IL-6 and IL-8 concentrations were significantly higher in the MEC-hypothermia group than in the MEC-normothermia (p < 0.001, p < 0.001, p = 0.001, p < 0.001, respectively) and sham-controlled groups (p < 0.001, p < 0.001, p < 0.001, p < 0.001, respectively). CONCLUSION: Meconium-induced inflammatory cytokine production is affected by the body temperature control.
Asunto(s)
Hipotermia Inducida/métodos , Síndrome de Aspiración de Meconio/patología , Síndrome de Aspiración de Meconio/terapia , Neumonía/patología , Neumonía/terapia , Animales , Líquido del Lavado Bronquioalveolar/química , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Mediciones Luminiscentes/métodos , Pulmón/patología , Masculino , Síndrome de Aspiración de Meconio/metabolismo , Neumonía/metabolismo , Ratas Wistar , Reproducibilidad de los Resultados , Resultado del Tratamiento , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Abstract Purpose: To evaluate the effects of hypothermia treatment on meconium-induced inflammation. Methods: Fifteen rats were instilled with human meconium (MEC, 1.5 mL/kg, 65 mg/mL) intratracheally and ventilated for 3 hours. Eight rats that were ventilated and not instilled with meconium served as a sham group. In MEC-hypothermia group, the body temperature was lowered to 33±0.5°C. Analysis of the blood gases, interleukin (IL)-1β, IL-6, IL-8, and tumor necrosis factor (TNF)-α in bronchoalveolar lavage (BAL) fluid samples, and histological analyses of the lungs were performed. Results: The BAL fluid TNF-α, IL-1β, IL-6 and IL-8 concentrations were significantly higher in the MEC-hypothermia group than in the MEC-normothermia (p < 0.001, p < 0.001, p = 0.001, p < 0.001, respectively) and sham-controlled groups (p < 0.001, p < 0.001, p < 0.001, p < 0.001, respectively). Conclusion: Meconium-induced inflammatory cytokine production is affected by the body temperature control.
Asunto(s)
Animales , Masculino , Neumonía/patología , Síndrome de Aspiración de Meconio/patología , Síndrome de Aspiración de Meconio/terapia , Hipotermia Inducida/métodos , Neumonía/metabolismo , Neumonía/terapia , Ensayo de Inmunoadsorción Enzimática , Líquido del Lavado Bronquioalveolar/química , Síndrome de Aspiración de Meconio/metabolismo , Reproducibilidad de los Resultados , Interleucina-8/metabolismo , Interleucina-6/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Resultado del Tratamiento , Ratas Wistar , Modelos Animales de Enfermedad , Interleucina-1beta/metabolismo , Mediciones Luminiscentes/métodos , Pulmón/patologíaRESUMEN
OBJECTIVE: Meconium aspiration syndrome (MAS) is a common cause of neonatal morbidity and mortality. Incomplete understanding of the pathogenesis of MAS has hindered the development of specific therapies. We hypothesized that activation of Toll-like receptors (TLRs) might play a role in the pathogenesis of MAS. The present study evaluated the expression of TLR 1, 4, 7, 8 and 9 in neonates with MAS. STUDY DESIGN: The study included 39 neonates with MAS and 17 healthy gestational age-matched neonates as controls. Neonates with maternal chorioamnionitis, perinatal asphyxia, sepsis and congenital malformations were excluded. Good-quality total RNA from umbilical cord blood was reverse transcribed to prepare cDNA using Bio-Rad reverse transcription kit. This cDNA was used to study the expression status of TLR 1, 4, 7, 8 and 9 by real-time quantitative polymerase chain reaction. RESULTS: Compared with controls, TLR1 and TLR4 were highly expressed, TLR9 was moderately expressed, TLR7 was weakly expressed and TLR8 expression was neutral in neonates with MAS. Within the MAS group, no difference in TLR expression was observed with respect to consistency of meconium, severity of the disease, oxygenation index and outcome. CONCLUSION: There is activation of TLRs in neonates with MAS. We speculate that these TLRs probably act as endogenous ligands for various components of meconium that initiate the inflammatory cascade of MAS and contribute to its pathogenesis.
Asunto(s)
Inflamación/metabolismo , Síndrome de Aspiración de Meconio/metabolismo , Receptores Toll-Like/metabolismo , Biomarcadores/sangre , Estudios de Casos y Controles , Femenino , Sangre Fetal , Edad Gestacional , Humanos , Recién Nacido , Masculino , Síndrome de Aspiración de Meconio/sangre , Estudios Prospectivos , Transducción de Señal , Receptores Toll-Like/sangreRESUMEN
Acute lung injury in the preterm newborns can originate from prematurity of the lung and insufficient synthesis of pulmonary surfactant. This situation is known as respiratory distress syndrome (RDS). In the term neonates, the respiratory insufficiency is related to a secondary inactivation of the pulmonary surfactant, for instance, by action of endotoxins in bacterial pneumonia or by effects of aspirated meconium. The use of experimental models of the mentioned situations provides new information on the pathophysiology of these disorders and offers unique possibility to test novel therapeutic approaches in the conditions which are very similar to the clinical syndromes. Herewith we review the advantages and limitations of the use of experimental models of RDS and meconium aspiration syndrome (MAS) and their value for clinics.
Asunto(s)
Lesión Pulmonar Aguda/fisiopatología , Modelos Animales de Enfermedad , Síndrome de Aspiración de Meconio/fisiopatología , Síndrome de Dificultad Respiratoria del Recién Nacido/fisiopatología , Lesión Pulmonar Aguda/metabolismo , Animales , Humanos , Recién Nacido , Síndrome de Aspiración de Meconio/metabolismo , Síndrome de Dificultad Respiratoria del Recién Nacido/metabolismoRESUMEN
Meconium aspiration syndrome (MAS) triggers inflammatory and oxidative pathways which can inactivate both pulmonary surfactant and therapeutically given exogenous surfactant. Glucocorticoid budesonide added to exogenous surfactant can inhibit inflammation and thereby enhance treatment efficacy. Neonatal meconium (25 mg/ml, 4 ml/kg) was administered intratracheally (i.t.) to rabbits. When the MAS model was prepared, animals were treated with budesonide i.t. (Pulmicort, 0.25 mg/kg, M+B); with surfactant lung lavage (Curosurf®, 10 ml/kg, 5 mg phospholipids/ml, M+S) followed by undiluted Curosurf® i.t. (100 mg phospholipids/kg); with combination of budesonide and surfactant (M+S+B); or were untreated (M); or served as controls with saline i.t. instead of meconium (C). Animals were oxygen-ventilated for additional 5 h. Cell counts in the blood and bronchoalveolar lavage fluid (BAL), lung edema formation (wet/dry weight ratio), oxidative damage of lipids/ proteins and inflammatory expression profiles (IL-2, IL-6, IL-13, TNF-alpha) in the lung homogenate and plasma were determined. Combined surfactant+budesonide therapy was the most effective in reduction of neutrophil counts in BAL, oxidative damage, levels and mRNA expression of cytokines in the lung, and lung edema formation compared to untreated animals. Curosurf fortified with budesonide mitigated lung inflammation and oxidative modifications what indicate the perspectives of this treatment combination for MAS therapy.
Asunto(s)
Budesonida/administración & dosificación , Modelos Animales de Enfermedad , Mediadores de Inflamación/metabolismo , Síndrome de Aspiración de Meconio/metabolismo , Estrés Oxidativo/efectos de los fármacos , Surfactantes Pulmonares/administración & dosificación , Animales , Antiinflamatorios/administración & dosificación , Quimioterapia Combinada , Femenino , Mediadores de Inflamación/antagonistas & inhibidores , Peroxidación de Lípido/efectos de los fármacos , Peroxidación de Lípido/fisiología , Masculino , Síndrome de Aspiración de Meconio/tratamiento farmacológico , Estrés Oxidativo/fisiología , ConejosAsunto(s)
Hipotermia Inducida/métodos , Síndrome de Aspiración de Meconio/terapia , Australia , Brasil , Estudios de Cohortes , Unión Europea , Femenino , Humanos , Recién Nacido , Tiempo de Internación/estadística & datos numéricos , Masculino , Síndrome de Aspiración de Meconio/metabolismo , Oxígeno/metabolismo , Respiración Artificial/estadística & datos numéricos , Estudios Retrospectivos , Factores de Riesgo , Resultado del Tratamiento , Estados UnidosRESUMEN
Neonatal meconium aspiration syndrome (MAS) can be treated by exogenous surfactant (S). However, aspirated meconium initiates local inflammation and oxidation which may inactivate surfactant and reduce its action. This experimental study estimated whether combined use of surfactant and the antioxidant N-acetylcysteine (NAC) can enhance effectiveness of therapy. Meconium-instilled rabbits were non-treated (M), treated with monotherapies (M + S, M + NAC), combined therapy (M + S + NAC), or received saline instead of meconium (controls, C). Surfactant therapy consisted of two lung lavages (BAL) with diluted Curosurf (5 mg phospholipids/ml, 10 ml/kg) followed by undiluted Curosurf (100 mg phospholipids/kg). N-acetylcysteine (Acc Injekt, 10 mg/kg) was given intravenously in M + S + NAC group 10 min after surfactant therapy. Animals were oxygen-ventilated for additional 5 h. Then, differential white cell count in the blood (WBC) was determined. Left lung was saline-lavaged and differential cell count in BAL was determined. In right lung tissue, wet/dry weight ratio, oxidation markers (TBARS, 3NT) and interleukines (IL-2, IL-6, IL-13, and TNFα) using ELISA and RT-PCR were estimated. Combined S + NAC therapy significantly decreased W/D ratio, TBARS, 3NT, and IL, whereas the effect of monotherapies (either S or NAC) was less obvious. In conclusion, addition of NAC to surfactant treatment may enhance the therapeutic outcome in MAS.
Asunto(s)
Acetilcisteína/uso terapéutico , Antiinflamatorios/uso terapéutico , Síndrome de Aspiración de Meconio/tratamiento farmacológico , Surfactantes Pulmonares/uso terapéutico , Acetilcisteína/farmacología , Animales , Antiinflamatorios/farmacología , Citocinas/metabolismo , Modelos Animales de Enfermedad , Quimioterapia Combinada , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Síndrome de Aspiración de Meconio/metabolismo , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/fisiología , Surfactantes Pulmonares/farmacología , Conejos , Resultado del TratamientoRESUMEN
OBJECTIVES: To evaluate the combined effects of surfactant lavage and intratracheally instillation of budesonide on meconium-injured piglet lungs. DESIGN: A prospective, randomized, animal model study. SETTING: An experimental laboratory. SUBJECTS: Twenty-four anesthetized and mechanically ventilated newborn piglets. INTERVENTIONS: Human meconium slurry was intratracheally instilled into piglet lungs to induce lung injury. The injured piglets were randomly assigned to either the sham treatment group (control) or one of the three therapeutic groups: the intratracheally instilled budesonide (Bud) group, the bronchoalveolar lavage with diluted surfactant (dsBAL) group, and the combination therapy of Bud and dsBAL (dsBAL + Bud) group. MEASUREMENTS AND MAIN RESULTS: Cardiopulmonary profiles were measured hourly. Proinflammatory cytokine (interleukin-1ß, interleukin-6, and interleukin-8) levels in bronchoalveolar lavage fluid were measured. Finally, the pulmonary histology of the experimental subjects was examined at the end of experiments. Both of the lavaged groups (dsBAL and dsBAL + Bud) showed significantly better oxygenation than those that had not undergone lavage (control and Bud) (p < 0.05). The dsBAL + Bud group showed a significantly higher lung compliance and required a significantly lower peak inspiratory pressure during the experimental periods than the other three groups (p < 0.05). All treatment groups had significantly lower concentrations of interleukin-1ß concentration in the bronchoalveolar lavage fluid than the control group (p < 0.05). The dsBAL + Bud group also had a significantly lower interleukin-6 concentration in the bronchoalveolar lavage fluid (p< 0.05), as well as a significantly lower lung injury score based on pulmonary histology than the control group (p < 0.05). CONCLUSIONS: Therapeutic bronchoalveolar lavage with diluted surfactant followed by intratracheal instillation of budesonide has a synergistic and beneficial effect when treating severe meconium-injured newborn piglet lungs.
Asunto(s)
Antiinflamatorios/uso terapéutico , Lavado Broncoalveolar , Budesonida/uso terapéutico , Síndrome de Aspiración de Meconio/terapia , Surfactantes Pulmonares/uso terapéutico , Animales , Biomarcadores/metabolismo , Líquido del Lavado Bronquioalveolar/química , Terapia Combinada , Citocinas/metabolismo , Humanos , Instilación de Medicamentos , Masculino , Síndrome de Aspiración de Meconio/metabolismo , Síndrome de Aspiración de Meconio/patología , Estudios Prospectivos , Distribución Aleatoria , Respiración Artificial , Porcinos , Resultado del TratamientoRESUMEN
Since inflammation and oxidative stress are fundamental in the pathophysiology of neonatal meconium aspiration syndrome (MAS), various anti-inflammatory drugs have been used in experimental and clinical studies on MAS. This pilot study evaluated therapeutic potential of N-acetylcysteine in modulation of meconium-induced inflammation and oxidative lung injury. Oxygen-ventilated adult rabbits were intratracheally given 4 ml/kg of meconium (25 mg/ml) or saline (Sal, n = 6). Thirty minutes later, meconium-instilled animals were treated with intravenous N-acetylcysteine (10 mg/kg, Mec + NAC, n=6) or were non-treated (Mec, n = 6). All animals were oxygen-ventilated for additional 5 hours. Total and differential blood leukocyte counts were determined at baseline, and at 1, 3 and 5 h of the treatment. After sacrificing animals, left lung was saline-lavaged and total and differential cell counts in the bronchoalveolar lavage fluid were determined. Right lung was used for biochemical analyses and for estimation of wet-dry weight ratio. In lung tissue homogenate, thiobarbituric acid-reactive substances (TBARS), dityrosine, lysine-lipid peroxidation (LPO) products, and total antioxidant status (TAS) were detected. In isolated lung mitochondria, TBARS, dityrosine, lysine-LPO products, thiol group content, conjugated dienes, and activity of cytochrome c oxidase were estimated. To evaluate systemic effects of meconium instillation and NAC treatment, TBARS and TAS were determined also in plasma. To evaluate participation of eosinophils in the meconium-induced inflammation, eosinophil cationic protein (ECP) was detected in plasma and lung homogenate. Meconium instillation increased oxidation markers and ECP in the lung and decreased TAS (all P<0.05). NAC treatment reduced ECP and oxidation markers (all P<0.05, except of dityrosine in homogenate and conjugated dienes in mitochondria) and prevented a decrease in TAS (P<0.01) in lung homogenate compared to Mec group. In plasma, NAC decreased TBARS (P<0.001) and ECP, and increased TAS (both P<0.05) compared to Mec group. Concluding, N-acetylcysteine diminished meconium-induced inflammation and oxidative lung injury.
Asunto(s)
Acetilcisteína/farmacología , Antioxidantes/farmacología , Lesión Pulmonar/prevención & control , Pulmón/efectos de los fármacos , Síndrome de Aspiración de Meconio/prevención & control , Meconio , Estrés Oxidativo/efectos de los fármacos , Neumonía/prevención & control , Factores de Edad , Animales , Biomarcadores/metabolismo , Modelos Animales de Enfermedad , Humanos , Recién Nacido , Mediadores de Inflamación/metabolismo , Leucocitos/efectos de los fármacos , Leucocitos/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Pulmón/inmunología , Pulmón/metabolismo , Lesión Pulmonar/inducido químicamente , Lesión Pulmonar/inmunología , Lesión Pulmonar/metabolismo , Síndrome de Aspiración de Meconio/inducido químicamente , Síndrome de Aspiración de Meconio/inmunología , Síndrome de Aspiración de Meconio/metabolismo , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Neumonía/inducido químicamente , Neumonía/inmunología , Neumonía/metabolismo , Edema Pulmonar/metabolismo , Edema Pulmonar/prevención & control , Conejos , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo , Factores de TiempoRESUMEN
Meconium aspiration in newborns causes lung inflammation and injury, which may lead to meconium aspiration syndrome (MAS). In this study, the effect of the antioxidant N-acetylcysteine on respiratory and inflammatory parameters were studied in a model of MAS. Oxygen-ventilated rabbits were intratracheally given 4 mL/kg of meconium (25 mg/mL) or saline. Thirty minutes later, meconium-instilled animals were administered N-acetylcysteine (10 mg/kg; i.v.), or were left without treatment. The animals were oxygen-ventilated for additional 5 h. Ventilatory pressures, oxygenation, right-to-left pulmonary shunts, and leukocyte count were measured. At the end of experiment, trachea and lung were excised. The left lung was saline-lavaged and a total and differential count of cells in bronchoalveolar lavage fluid (BAL) was determined. Right lung tissue strips were used for detection of lung edema (expressed as wet/dry weight ratio) and peroxidation (expressed by thiobarbituric acid-reactive substances, TBARS). In lung and tracheal strips, airway reactivity to acetylcholine was measured. In addition, TBARS and total antioxidant status were determined in the plasma. Meconium instillation induced polymorphonuclear-derived inflammation and oxidative stress. N-acetylcysteine improved oxygenation, reduced lung edema, decreased polymorphonuclears in BAL fluid, and diminished peroxidation and meconium-induced airway hyperreactivity compared with untreated animals. In conclusion, N-acetylcysteine effectively improved lung functions in an animal model of MAS.
Asunto(s)
Acetilcisteína/farmacología , Lesión Pulmonar Aguda/tratamiento farmacológico , Antiinflamatorios/farmacología , Antioxidantes/farmacología , Síndrome de Aspiración de Meconio/tratamiento farmacológico , Edema Pulmonar/tratamiento farmacológico , Lesión Pulmonar Aguda/inducido químicamente , Lesión Pulmonar Aguda/metabolismo , Lesión Pulmonar Aguda/fisiopatología , Animales , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Modelos Animales de Enfermedad , Humanos , Recién Nacido , Inyecciones Intravenosas , Intubación Intratraqueal , Recuento de Leucocitos , Peroxidación de Lípido/efectos de los fármacos , Pulmón/efectos de los fármacos , Pulmón/fisiopatología , Meconio , Síndrome de Aspiración de Meconio/metabolismo , Síndrome de Aspiración de Meconio/fisiopatología , Estrés Oxidativo , Edema Pulmonar/inducido químicamente , Edema Pulmonar/metabolismo , Edema Pulmonar/fisiopatología , Conejos , Respiración Artificial , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo , Tráquea/efectos de los fármacos , Tráquea/fisiopatologíaRESUMEN
Meconium aspiration syndrome (MAS) is meconium-induced respiratory failure of newborns associated with activation of inflammatory and oxidative pathways. For severe MAS, exogenous surfactant treatment is used which improves respiratory functions but does not treat the inflammation. Oxidative process can lead to later surfactant inactivation; hence, surfactant combination with antioxidative agent may enhance the therapeutic effect. Young New Zealand rabbits were instilled by meconium suspension and treated by surfactant alone, N-acetylcysteine (NAC) alone or by their combination and oxygen-ventilated for 5 h. Blood samples were taken before and 30 min after meconium application and 30 min, 1, 3 and 5 h after the treatment for evaluating of oxidative damage, total leukocyte count, leukocyte differential count and respiratory parameters. Leukocyte differential was assessed also in bronchoalveolar lavage fluid. NAC alone had only mild therapeutic effect on MAS. However, the combination of NAC and surfactant facilitated rapid onset of therapeutic effect in respiratory parameters (oxygenation index, PaO(2)/FiO(2)) compared to surfactant alone and was the only treatment which prevented neutrophil migration into the lungs, oxidative damage and lung edema. Moreover, NAC suppressed IL-8 and IL-beta formation and thus seems to be favorable agent for improving surfactant therapy in MAS.
Asunto(s)
Acetilcisteína/uso terapéutico , Expectorantes/uso terapéutico , Síndrome de Aspiración de Meconio/prevención & control , Surfactantes Pulmonares/uso terapéutico , Acetilcisteína/farmacología , Animales , Animales Recién Nacidos , Líquido del Lavado Bronquioalveolar/inmunología , Ensayos de Migración de Leucocitos , Citocinas/metabolismo , Evaluación Preclínica de Medicamentos , Quimioterapia Combinada , Edema/prevención & control , Expectorantes/farmacología , Recuento de Leucocitos , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Síndrome de Aspiración de Meconio/inmunología , Síndrome de Aspiración de Meconio/metabolismo , Proyectos Piloto , Surfactantes Pulmonares/farmacología , Conejos , Distribución Aleatoria , Pruebas de Función Respiratoria , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo , Tirosina/análogos & derivados , Tirosina/metabolismoRESUMEN
BACKGROUND: Meconium-stained amniotic fluid (MSAF) represents the passage of fetal colonic content into the amniotic cavity. Meconium aspiration syndrome (MAS) is a complication that occurs in a subset of infants with MSAF. Secreted phospholipase A2 (sPLA2) is detected in meconium and is implicated in the development of MAS. The purpose of this study was to determine if sPLA2 concentrations are increased in the amniotic fluid of women in spontaneous labor at term with MSAF. MATERIALS AND METHODS: This was a cross-sectional study of patients in spontaneous term labor who underwent amniocentesis (n = 101). The patients were divided into two study groups: (1) MSAF (n = 61) and (2) clear fluid (n = 40). The presence of bacteria and endotoxin as well as interleukin-6 (IL-6) and sPLA2 concentrations in the amniotic fluid were determined. Statistical analyses were performed to test for normality and bivariate analysis. The Spearman correlation coefficient was used to study the relationship between sPLA2 and IL-6 concentrations in the amniotic fluid. RESULTS: Patients with MSAF have a higher median sPLA2 concentration (ng/mL) in amniotic fluid than those with clear fluid [1.7 (0.98-2.89) versus 0.3 (0-0.6), p < 0.001]. Among patients with MSAF, those with either microbial invasion of the amniotic cavity (MIAC, defined as presence of bacteria in the amniotic cavity), or bacterial endotoxin had a significantly higher median sPLA2 concentration (ng/mL) in amniotic fluid than those without MIAC or endotoxin [2.4 (1.7-6.0) versus 1.7 (1.3-2.5), p < 0.05]. There was a positive correlation between sPLA2 and IL-6 concentrations in the amniotic fluid (Spearman Rho = 0.3, p < 0.05). CONCLUSION: MSAF that contains bacteria or endotoxin has a higher concentration of sPLA2, and this may contribute to induce lung inflammation when meconium is aspirated before birth.
Asunto(s)
Líquido Amniótico/metabolismo , Síndrome de Aspiración de Meconio/etiología , Fosfolipasas A2 Secretoras/metabolismo , Amniocentesis , Líquido Amniótico/química , Líquido Amniótico/microbiología , Biomarcadores/metabolismo , Estudios Transversales , Endotoxinas/análisis , Femenino , Humanos , Recién Nacido , Interleucina-6/metabolismo , Síndrome de Aspiración de Meconio/metabolismo , Embarazo , Nacimiento a TérminoRESUMEN
Surfactant dysfunction in meconium aspiration syndrome (MAS) is caused by meconium components, by plasma proteins leaking through the injured alveolocapillary membrane and by substances originated in meconium-induced inflammation. Surfactant inactivation in MAS may be diminished by several ways. Firstly, aspirated meconium should be removed from the lungs to decrease concentrations of meconium inhibitors coming into the contact with surfactant in the alveolar compartment. Once the endogenous surfactant becomes inactivated, components of surfactant should be substituted by exogenous surfactant at a sufficient dose, and surfactant administration should be repeated, if oxygenation remains compromised. To delay the inactivation by inhibitors, exogenous surfactants may be enriched with surfactant proteins, phospholipids, or other substances such as polymers. Finally, to diminish an adverse action of products of meconium-induced inflammation on both endogenous and exogenously delivered surfactant, anti-inflammatory drugs may be administered. A combined therapeutic approach may result in better outcome in patients with MAS and in lower costs of treatment.
Asunto(s)
Síndrome de Aspiración de Meconio/tratamiento farmacológico , Síndrome de Aspiración de Meconio/fisiopatología , Surfactantes Pulmonares/metabolismo , Surfactantes Pulmonares/uso terapéutico , Animales , Antiinflamatorios/uso terapéutico , Humanos , Recién Nacido , Inflamación/tratamiento farmacológico , Inflamación/etiología , Inflamación/fisiopatología , Síndrome de Aspiración de Meconio/metabolismoRESUMEN
OBJECTIVE: The aim of this study was to compare cord blood S100B levels and cord blood gas parameters of term infants with meconium-stained amniotic fluid (MSAF) to those infants with clear amniotic fluid. METHODS: Term pregnant women at an active phase of labor and having MSAF were defined as the study group (n = 35) and pregnant women with clear amniotic fluid, and matched for age, parity, and gestational age were defined as the control group (n = 35). Cord blood S100B levels and gas parameters were measured. RESULTS: LogS100B values of study and control groups were 2.40 ± 0.21 and 2.43 ± 0.29 pg/ml, respectively. The difference was not statistically significant (p = 0.675). LogS100B levels slightly increased as meconium thickened. (2.32 ± 0.16, 2.41 ± 0.17, and 2.44 ± 0.28 pg/ml, respectively). However, no difference was found between groups (p = 0.438). Moreover, the study group had a statistically lower HCO(3) level (21.80 vs 23.60 mmol/l) and a higher rate of base deficit (4.85 vs 3.25 mmol/l) than the control group. However, median HCO(3) and base deficit values were within normal limits in both groups. CONCLUSION: The present study showed that cord blood S100B levels of infants born through MSAF were not different from those with clear amniotic fluid. This finding suggests that MSAF, regardless of its thickness, may not be related to brain damage in low risk term pregnancies.
Asunto(s)
Líquido Amniótico/metabolismo , Sangre Fetal/química , Meconio/metabolismo , Factores de Crecimiento Nervioso/sangre , Proteínas S100/sangre , Nacimiento a Término/sangre , Adulto , Bicarbonatos/análisis , Bicarbonatos/sangre , Análisis de los Gases de la Sangre , Estudios de Casos y Controles , Femenino , Sangre Fetal/metabolismo , Humanos , Recién Nacido , Síndrome de Aspiración de Meconio/sangre , Síndrome de Aspiración de Meconio/diagnóstico , Síndrome de Aspiración de Meconio/metabolismo , Factores de Crecimiento Nervioso/análisis , Embarazo , Factores de Riesgo , Subunidad beta de la Proteína de Unión al Calcio S100 , Proteínas S100/análisis , Nacimiento a Término/fisiología , Adulto JovenRESUMEN
To examine the effects of pentoxifylline (PTX) on regional pulmonary and systemic inflammation after meconium aspiration, we studied 26 anesthetized and ventilated adult rats for 3 hours. Seventeen rats were instilled with human meconium (1.5 mL/kg, 65 mg/mL) intratracheally. After instillation of meconium, PTX (20 mg/kg, i.a.; n = 9) or saline (n = 8) was given to the subjects. Nine rats that were ventilated and not instilled with meconium served as sham group. Meconium instillation resulted in increased bronchoalveolar lavage (BAL) fluid tumor necrosis factor-α (TNF-α; P = 0.004 and P = 0.002, respectively), protein (P = 0.005 and P = 0.001, respectively) levels, and arterial oxygenation index (OI) in PTX and saline groups. PTX treatment prevented the increase of BAL fluid TNF-α, protein concentrations, and OI in the meconium-instilled lungs but had no statistically significant effect. These results indicate that meconium aspiration induces severe inflammation in the lung. PTX treatment affects the TNF-α production in the lungs and it may attenuate meconium-induced derangements.
Asunto(s)
Síndrome de Aspiración de Meconio/tratamiento farmacológico , Pentoxifilina/farmacología , Neumonía/tratamiento farmacológico , Animales , Arterias/efectos de los fármacos , Arterias/metabolismo , Líquido del Lavado Bronquioalveolar/química , Modelos Animales de Enfermedad , Humanos , Recién Nacido , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Pulmón/patología , Meconio/metabolismo , Síndrome de Aspiración de Meconio/metabolismo , Síndrome de Aspiración de Meconio/patología , Neumonía/metabolismo , Neumonía/patología , Ratas , Ratas Wistar , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
BACKGROUND: Surfactant lavage has been used to remove meconium debris in meconium aspiration syndrome (MAS), but the influence of surfactant lavage on pro-inflammatory cytokines and cellular apoptosis is unclear. The aim of this study was to investigate the response of pro-inflammatory cytokine and the influence on alveolar cellular apoptosis using therapeutic bronchoalveolar lavage with diluted surfactant to treat MAS. METHODS: Twelve newborn piglets were anesthetized, intubated via tracheostomy, and artificially ventilated. MAS was induced by intratracheal instillation of 3-5 mL/kg of 20% human meconium. The piglets were then randomly assigned to a surfactant lavage group (n= 6) or a control group (n= 6). Piglets in the lavage group received bronchoalveolar lavage with 30 mL/kg diluted surfactant (5 mg/mL) in two aliquots. Cardiopulmonary parameters were monitored continuously. Serum was obtained hourly to measure concentrations of pro-inflammatory cytokines, including interleukin (IL)-I beta, IL-6, and tumor necrosis factor alpha. Lung tissue was histologically examined after experiments, and terminal deoxynucleotidyl transferase-mediated nick-end labeling assay for apoptotic cell death was also performed. RESULTS: The animals in the lavage group displayed significantly better gas exchange and lower serum concentrations of IL-1 beta than the animals in the control group (P < 0.05). The number of apoptotic cells in lung tissues was significantly lower in the lavage group than the control group, and also in the nondependent than the dependent site. CONCLUSION: Therapeutic surfactant lavage improves oxygenation, decreases production of systemic pro-inflammatory cytokine IL-1 beta, and alleviates the severity of lung cell apoptosis in newborn piglets with experimentally-induced MAS.