RESUMO
We employed an early training exercise program, immediately after recovery from surgery, and before severe cardiac hypertrophy, to study the underlying mechanism involved with the amelioration of cardiac dysfunction in aortic stenosis (AS) rats. As ET induces angiogenesis and oxygen support, we aimed to verify the effect of exercise on myocardial lipid metabolism disturbance. Wistar rats were divided into Sham, trained Sham (ShamT), AS and trained AS (AST). The exercise consisted of 5-week sessions of treadmill running for 16 weeks. Statistical analysis was conducted by anova or Kruskal-Wallis test and Goodman test. A global correlation between variables was also performed using a two-tailed Pearson's correlation test. AST rats displayed a higher functional capacity and a lower cardiac remodelling and dysfunction when compared to AS, as well as the myocardial capillary rarefaction was prevented. Regarding metabolic properties, immunoblotting and enzymatic assay raised beneficial effects of exercise on fatty acid transport and oxidation pathways. The correlation assessment indicated a positive correlation between variables of angiogenesis and FA utilisation, as well as between metabolism and echocardiographic parameters. In conclusion, early exercise improves exercise tolerance and attenuates cardiac structural and functional remodelling. In parallel, exercise attenuated myocardial capillary and lipid metabolism derangement in rats with aortic stenosis-induced heart failure.
Assuntos
Estenose da Valva Aórtica , Insuficiência Cardíaca , Condicionamento Físico Animal , Ratos , Animais , Ratos Wistar , Metabolismo dos Lipídeos , Insuficiência Cardíaca/metabolismoRESUMO
Background: Patients with congenital diaphragmatic hernia (CDH) have a short postnatal period of ventilatory stability called the honeymoon period, after which changes in pulmonary vascular reactivity result in pulmonary hypertension. However, the mechanisms involved are still unknown. The aim of this study was to evaluate mechanical ventilation's effect in the honeymoon period on VEGF, VEGFR-1/2 and eNOS expression on experimental CDH in rats. Materials and Methods: Neonates whose mothers were not exposed to nitrofen formed the control groups (C) and neonates with left-sided defects formed the CDH groups (CDH). Both were subdivided into non-ventilated and ventilated for 30, 60, and 90 min (n = 7 each). The left lungs (n = 4) were evaluated by immunohistochemistry of the pulmonary vasculature (media wall thickness), VEGF, VEGFR-1/2 and eNOS. Western blotting (n = 3) was performed to quantify the expression of VEGF, VEGFR-1/2 and eNOS. Results: CDH had lower biometric parameters than C. Regarding the pulmonary vasculature, C showed a reduction in media wall thickness with ventilation, while CDH presented reduction with 30 min and an increase with the progression of the ventilatory time (honeymoon period). CDH and C groups showed different patterns of VEGF, VEGFR-1/2 and eNOS expressions. The receptors and eNOS findings were significant by immunohistochemistry but not by western blotting, while VEGF was significant by western blotting but not by immunohistochemistry. Conclusion: VEGF, its receptors and eNOS were altered in CDH after mechanical ventilation. These results suggest that the VEGF-NO pathway plays an important role in the honeymoon period of experimental CDH.
RESUMO
The survival rate of newborns with gastroschisis (GS) has been increasing in the past decades; however, the morbidity continues to be high, mainly related to the length of hospitalization due to disturbances of motility, absorption, and risk of intestinal infections. The development of basic research with the creation of experimental models has provided enormous advances in the understanding of the pathophysiology of the disease. These models allowed the study of the target genes involved in the embryology of the defect, the influence of the amniotic fluid, and the use of drugs and fetal therapies in an attempt to reduce the intestinal damage and to provide a rapid return of intestinal motility. Our aim was to describe the main GS models and the translational, historical impact of these research advances on the disease.
Assuntos
Modelos Animais de Doenças , Gastrosquise , Pesquisa Translacional Biomédica/métodos , Animais , Gastrosquise/etiologia , Gastrosquise/fisiopatologia , Gastrosquise/terapia , HumanosRESUMO
BACKGROUND: Biliary cirrhosis is associated with hepatopulmonary syndrome (HPS), which is related to increased posttransplant morbidity and mortality. AIMS: This study aims to analyze the pathophysiology of biliary cirrhosis and the onset of HPS. METHODS: Twenty-one-day-old Wistar rats were subjected to common bile duct ligation and were allocated to two groups: group A (killed 2, 3, 4, 5, or 6 weeks after biliary obstruction) and group B (subjected to biliodigestive anastomosis 2, 3, 4, 5, or 6 weeks after the first procedure and killed 3 weeks later). At the killing, arterial blood was collected for the analyses, and samples from the liver and lungs were collected for histologic and molecular analyses. The gasometric parameters as well as the expression levels of ET-1, eNOS, and NOS genes in the lung tissue were evaluated. RESULTS: From a total of 42 blood samples, 15 showed hypoxemia (pO2 < 85 mmHg) and 17 showed an increased oxygen gradient [p (A-a) O2 > 18 mmHg]. The liver histology revealed increased ductular proliferation after common bile duct ligation, and reconstruction of bile flow promoted decreased ductular proliferation 5 and 6 weeks post-common bile duct ligation. Pulmonary alterations consisted of decreased parenchymal airspace and increased medial wall thickness. Biliary desobstruction promoted transitory improvements 5 weeks after biliary obstruction (increased parenchymal airspace and decreased MWT-p = 0.003 and p = 0.004, respectively) as well as increased endothelin expression levels (p = 0.009). CONCLUSIONS: The present model showed lung tissue alterations promoted by biliary obstruction. The biliodigestive anastomosis had no clear direct effects on these alterations.
Assuntos
Ductos Biliares/patologia , Modelos Animais de Doenças , Síndrome Hepatopulmonar/patologia , Cirrose Hepática Biliar/patologia , Anastomose Cirúrgica/métodos , Animais , Ductos Biliares/cirurgia , Feminino , Síndrome Hepatopulmonar/sangue , Ligadura , Cirrose Hepática Biliar/sangue , Pulmão/patologia , Masculino , Ratos , Ratos WistarRESUMO
PURPOSE: To evaluate the vascular ventilatory response in different stages of lung development and to compare them to the neonates with congenital diaphragmatic hernia (CDH) in a rabbit model. METHODS: New Zealand rabbits were divided into 8 groups (n=5): E25, E27, E30, and CDH. All groups were ventilated on a FlexiVent (Scireq, Montreal, QC, Canada), compounding the other 4 groups. The CDH surgery was performed at E25 and the harvest at E30. Dynamic compliance (CRS), dynamic elastance (ERS) and dynamic resistance (RRS) were measured every 4 min/24 min. Median wall thickness (MWT) and airspace were measured. ANOVA Bonferroni tests were used to perform statistical analysis. Significance was considered when p<0.05. RESULTS: CRS was higher in E30 compared to all other groups (p<0.05). CRS and RRS of CDH and E27 were similar and were higher in E25 (p<0.05). MWT was decreased according to the gestational age, was increased in E27V and E30V (p<0.05) and decreased in CDHV (p<0.05), airspace was decreased in E25 and increased in all ventilated groups (p<0.05). CONCLUSIONS: The ventilation response of congenital diaphragmatic hernia is like the pseudoglandular stage of the lung development. These findings add information about the physiology of pulmonary ventilation in CDH.
Assuntos
Hérnias Diafragmáticas Congênitas/fisiopatologia , Pulmão/crescimento & desenvolvimento , Mecânica Respiratória/fisiologia , Resistência das Vias Respiratórias , Animais , Animais Recém-Nascidos , Diafragma/cirurgia , Modelos Animais de Doenças , Hérnias Diafragmáticas Congênitas/etiologia , Pulmão/irrigação sanguínea , Pulmão/fisiopatologia , Coelhos , Testes de Função Respiratória , Capacidade Pulmonar TotalRESUMO
Purpose: To evaluate the vascular ventilatory response in different stages of lung development and to compare them to the neonates with congenital diaphragmatic hernia (CDH) in a rabbit model. Methods: New Zealand rabbits were divided into 8 groups (n=5): E25, E27, E30, and CDH. All groups were ventilated on a FlexiVent (Scireq, Montreal, QC, Canada), compounding the other 4 groups. The CDH surgery was performed at E25 and the harvest at E30. Dynamic compliance (CRS), dynamic elastance (ERS) and dynamic resistance (RRS) were measured every 4 min/24 min. Median wall thickness (MWT) and airspace were measured. ANOVA Bonferroni tests were used to perform statistical analysis. Significance was considered when p 0.05. Results: CRS was higher in E30 compared to all other groups (p 0.05). CRS and RRS of CDH and E27 were similar and were higher in E25 (p 0.05). MWT was decreased according to the gestational age, was increased in E27V and E30V (p 0.05) and decreased in CDHV (p 0.05), airspace was decreased in E25 and increased in all ventilated groups (p 0.05). Conclusions: The ventilation response of congenital diaphragmatic hernia is like the pseudoglandular stage of the lung development. These findings add information about the physiology of pulmonary ventilation in CDH.(AU)
Assuntos
Animais , Coelhos , Hérnias Diafragmáticas Congênitas/veterinária , Pulmão/crescimento & desenvolvimento , Ventilação Pulmonar/fisiologia , Modelos AnimaisRESUMO
Abstract Purpose: To evaluate the vascular ventilatory response in different stages of lung development and to compare them to the neonates with congenital diaphragmatic hernia (CDH) in a rabbit model. Methods: New Zealand rabbits were divided into 8 groups (n=5): E25, E27, E30, and CDH. All groups were ventilated on a FlexiVent (Scireq, Montreal, QC, Canada), compounding the other 4 groups. The CDH surgery was performed at E25 and the harvest at E30. Dynamic compliance (CRS), dynamic elastance (ERS) and dynamic resistance (RRS) were measured every 4 min/24 min. Median wall thickness (MWT) and airspace were measured. ANOVA Bonferroni tests were used to perform statistical analysis. Significance was considered when p<0.05. Results: CRS was higher in E30 compared to all other groups (p<0.05). CRS and RRS of CDH and E27 were similar and were higher in E25 (p<0.05). MWT was decreased according to the gestational age, was increased in E27V and E30V (p<0.05) and decreased in CDHV (p<0.05), airspace was decreased in E25 and increased in all ventilated groups (p<0.05). Conclusions: The ventilation response of congenital diaphragmatic hernia is like the pseudoglandular stage of the lung development. These findings add information about the physiology of pulmonary ventilation in CDH.
Assuntos
Animais , Coelhos , Mecânica Respiratória/fisiologia , Hérnias Diafragmáticas Congênitas/fisiopatologia , Pulmão/crescimento & desenvolvimento , Testes de Função Respiratória , Diafragma/cirurgia , Capacidade Pulmonar Total , Resistência das Vias Respiratórias , Modelos Animais de Doenças , Hérnias Diafragmáticas Congênitas/etiologia , Pulmão/fisiopatologia , Pulmão/irrigação sanguínea , Animais Recém-NascidosRESUMO
OBJECTIVES: The objective of the present study was to evaluate the protective effect of pre-conditioning treatment with laser light on hepatic injury in rats submitted to partial ischemia using mitochondrial function and liver fatty acid binding protein as markers. METHODS: Rats were divided into four groups (n=5): 1) Control, 2) Control + Laser, 3) Partial Ischemia and 4) Partial Ischemia + Laser. Ischemia was induced by clamping the hepatic pedicle of the left and middle lobes of the liver for 60 minutes. Laser light at 660 nm was applied to the liver immediately prior to the induction of ischemia at 22.5 J/cm2, with 30 seconds of illumination at five individual points. The animals were sacrificed after 30 minutes of reperfusion. Blood and liver tissues were collected for analysis of mitochondrial function, determination of malondialdehyde and analysis of fatty acid binding protein expression by Western blot. RESULTS: Mitochondrial function decreased in the Partial Ischemia group, especially during adenosine diphosphate-activated respiration (state 3), and the expression of fatty acid binding protein was also reduced. The application of laser light prevented bioenergetic changes and restored the expression of fatty acid binding protein. CONCLUSION: Prophylactic application of laser light to the livers of rats submitted to partial ischemia was found to have a protective effect in the liver, with normalization of both mitochondrial function and fatty acid binding protein tissue expression.
Assuntos
Proteínas de Ligação a Ácido Graxo/metabolismo , Precondicionamento Isquêmico/métodos , Fígado/irrigação sanguínea , Fígado/efeitos da radiação , Terapia com Luz de Baixa Intensidade/métodos , Traumatismo por Reperfusão/prevenção & controle , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Western Blotting , Proteínas de Ligação a Ácido Graxo/análise , Fígado/metabolismo , Malondialdeído/análise , Malondialdeído/efeitos da radiação , Membranas Mitocondriais/efeitos dos fármacos , Dilatação Mitocondrial/efeitos da radiação , Ratos Wistar , Reprodutibilidade dos TestesRESUMO
OBJECTIVES: The objective of the present study was to evaluate the protective effect of pre-conditioning treatment with laser light on hepatic injury in rats submitted to partial ischemia using mitochondrial function and liver fatty acid binding protein as markers. METHODS: Rats were divided into four groups (n=5): 1) Control, 2) Control + Laser, 3) Partial Ischemia and 4) Partial Ischemia + Laser. Ischemia was induced by clamping the hepatic pedicle of the left and middle lobes of the liver for 60 minutes. Laser light at 660 nm was applied to the liver immediately prior to the induction of ischemia at 22.5 J/cm2, with 30 seconds of illumination at five individual points. The animals were sacrificed after 30 minutes of reperfusion. Blood and liver tissues were collected for analysis of mitochondrial function, determination of malondialdehyde and analysis of fatty acid binding protein expression by Western blot. RESULTS: Mitochondrial function decreased in the Partial Ischemia group, especially during adenosine diphosphate-activated respiration (state 3), and the expression of fatty acid binding protein was also reduced. The application of laser light prevented bioenergetic changes and restored the expression of fatty acid binding protein. CONCLUSION: Prophylactic application of laser light to the livers of rats submitted to partial ischemia was found to have a protective effect in the liver, with normalization of both mitochondrial function and fatty acid binding protein tissue expression.
Assuntos
Animais , Traumatismo por Reperfusão/prevenção & controle , Precondicionamento Isquêmico/métodos , Terapia com Luz de Baixa Intensidade/métodos , Proteínas de Ligação a Ácido Graxo/metabolismo , Fígado/efeitos da radiação , Fígado/irrigação sanguínea , Aspartato Aminotransferases/sangue , Western Blotting , Reprodutibilidade dos Testes , Ratos Wistar , Alanina Transaminase/sangue , Membranas Mitocondriais/efeitos dos fármacos , Proteínas de Ligação a Ácido Graxo/análise , Fígado/metabolismo , Malondialdeído/análise , Malondialdeído/efeitos da radiação , Dilatação Mitocondrial/efeitos da radiaçãoRESUMO
BACKGROUND: Congenital diaphragmatic hernia (CDH) is associated with lung hypoplasia and pulmonary hypertension. Tracheal occlusion (TO) stimulates fetal lung growth and maturation and reverse vascular changes responsible for pulmonary hypertension, which are related to mechanisms involving nitric oxide (NO) in CDH. We aim to evaluate the effect of TO and ventilation on NO pathways. METHODS: Eight groups were created: (1) control; (2) control ventilated (CV); (3) CDH (CDH); (4) CDH ventilated (CDHV); (5) TO control; (6) TO ventilated; (7) TO + CDH; and (8) TO + CDH ventilated (CDHTOV). Fetuses were weighed, and volume ventilated for 30 min after harvested. Total lung weight and the ratio of total lung weight to body weight, thickness of the middle layer of the pulmonary arteriole, and the air space diameter were measured. The NO synthase inducible and NO synthase inducible were performed by immunohistochemistry and Western blotting. RESULTS: The total lung weight and the ratio of total lung weight to body weight decreased in animals with nitrofen and also after ventilation for all groups (P < 0.05). The thickness of the middle layer of the pulmonary arteriole decreased in all groups with TO when compared with controls (P < 0.001). The air space diameter decreased after ventilation in the CDHTOV compared to the TO + nitrofen-induced CDH (P < 0.001). Compared to nonventilated cohorts, NO synthase inducible increased in CV and TO ventilated (P < 0.001) and decreased in CDHV and CDHTOV (P < 0.001). NO synthase inducible increased in CV and CDHV (P < 0.001) and decreased in the TO control and CDHTOV (P < 0.001). CONCLUSIONS: TO and ventilation alter the NO pathway with possible implications in reducing the pulmonary hypertension in CDH.
Assuntos
Terapias Fetais , Hérnias Diafragmáticas Congênitas/terapia , Óxido Nítrico Sintase Tipo III/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Óxido Nítrico/metabolismo , Respiração Artificial , Oclusão Terapêutica , Animais , Biomarcadores/metabolismo , Western Blotting , Feminino , Terapias Fetais/métodos , Hérnias Diafragmáticas Congênitas/metabolismo , Hérnias Diafragmáticas Congênitas/fisiopatologia , Hipertensão Pulmonar/etiologia , Hipertensão Pulmonar/metabolismo , Hipertensão Pulmonar/prevenção & controle , Imuno-Histoquímica , Pulmão/embriologia , Pulmão/metabolismo , Óxido Nítrico Sintase/metabolismo , Tamanho do Órgão , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVES: Necrotizing enterocolitis is a severe multifactorial intestinal disorder that primarily affects preterm newborns, causing 20-40% mortality and morbidity. Intestinal fatty acid-binding protein has been reported to be a biomarker for the detection of intestinal injuries. Our aim was to assess intestinal tissue injury and the molecular expression of intestinal fatty acid-binding protein over time in a necrotizing enterocolitis model. METHODS: A total of 144 Newborn rats were divided into two groups: 1) Control, which received breastfeeding (n=72) and 2) Necrotizing Enterocolitis, which received formula feeding and underwent hypoxia and hypothermia (n=72). A total of six time points of ischemia (2 times a day for 3 days; 12 pups for each time point) were examined. Samples were collected for analysis of body weight, morphological and histological characteristics, intestinal weight, intestinal weight/body weight ratio, injury grade, and intestinal fatty acid-binding protein levels. RESULTS: Body and intestinal weights were lower in the Necrotizing Enterocolitis group than in the Control group (p<0.005 and p<0.0005, respectively). The intestinal weight/body weight ratio was higher in the Necrotizing Enterocolitis group than in the Control group (p<0.005) only at the sixth ischemia time point. The Necrotizing Enterocolitis group displayed higher expression of intestinal fatty acid-binding protein (p<0.0005) and showed greater tissue damage than the Control group. CONCLUSION: Intestinal fatty acid-binding protein was an efficient marker of ischemic injury to the intestine and a good correlation was demonstrated between the time of ischemic injury and the grade of intestinal injury.
Assuntos
Enterocolite Necrosante/metabolismo , Enterocolite Necrosante/patologia , Proteínas de Ligação a Ácido Graxo/metabolismo , Íleo/patologia , Animais , Animais Recém-Nascidos , Biomarcadores/análise , Western Blotting , Peso Corporal , Modelos Animais de Doenças , Proteínas de Ligação a Ácido Graxo/análise , Hipóxia/patologia , Íleo/irrigação sanguínea , Imuno-Histoquímica , Isquemia/patologia , Distribuição Aleatória , Ratos Sprague-Dawley , Valores de Referência , Índice de Gravidade de Doença , Fatores de TempoRESUMO
OBJECTIVES: Necrotizing enterocolitis is a severe multifactorial intestinal disorder that primarily affects preterm newborns, causing 20-40% mortality and morbidity. Intestinal fatty acid-binding protein has been reported to be a biomarker for the detection of intestinal injuries. Our aim was to assess intestinal tissue injury and the molecular expression of intestinal fatty acid-binding protein over time in a necrotizing enterocolitis model. METHODS: A total of 144 Newborn rats were divided into two groups: 1) Control, which received breastfeeding (n=72) and 2) Necrotizing Enterocolitis, which received formula feeding and underwent hypoxia and hypothermia (n=72). A total of six time points of ischemia (2 times a day for 3 days; 12 pups for each time point) were examined. Samples were collected for analysis of body weight, morphological and histological characteristics, intestinal weight, intestinal weight/body weight ratio, injury grade, and intestinal fatty acid-binding protein levels. RESULTS: Body and intestinal weights were lower in the Necrotizing Enterocolitis group than in the Control group (p<0.005 and p<0.0005, respectively). The intestinal weight/body weight ratio was higher in the Necrotizing Enterocolitis group than in the Control group (p<0.005) only at the sixth ischemia time point. The Necrotizing Enterocolitis group displayed higher expression of intestinal fatty acid-binding protein (p<0.0005) and showed greater tissue damage than the Control group. CONCLUSION: Intestinal fatty acid-binding protein was an efficient marker of ischemic injury to the intestine and a good correlation was demonstrated between the time of ischemic injury and the grade of intestinal injury.
Assuntos
Animais , Enterocolite Necrosante/metabolismo , Enterocolite Necrosante/patologia , Proteínas de Ligação a Ácido Graxo/metabolismo , Íleo/patologia , Valores de Referência , Fatores de Tempo , Índice de Gravidade de Doença , Peso Corporal , Imuno-Histoquímica , Biomarcadores/análise , Distribuição Aleatória , Western Blotting , Ratos Sprague-Dawley , Modelos Animais de Doenças , Proteínas de Ligação a Ácido Graxo/análise , Íleo/irrigação sanguínea , Isquemia/patologia , Animais Recém-Nascidos , Hipóxia/patologiaRESUMO
BACKGROUND/PURPOSE: Necrotizing enterocolitis (NEC) is a severe intestinal disease that primarily affects premature babies, leading to high mortality and morbidity. Probiotics represent an important alternative prophylaxis for NEC but its mechanism of action is poorly understood. Moreover, intestinal and liver-type fatty acid-binding proteins (I-FABP and L-FABP) may be utilized because markers of intestinal injury, including NEC. We aimed to evaluate the protection induced by the Lactobacillus acidophilus on the intestines of newborn rats submitted to experimental NEC using I-FABP and L-FABP as biomarkers. METHODS: Sprague-Dawley newborn rats were divided into three groups: (1) C (control)-breast-fed; (2) NEC-subjected to NEC protocol and (3) NECP-NEC+probiotic. Morphometric, intestinal lesion, immunohistochemistry and Western blotting analysis were performed. Statistical significant differences were considered when p<0.05. RESULTS: Intestinal weight was lower in NEC and NECP compared to C (p<0.05). Intestinal injury was lower in NECP compared to NEC. Prophylactic probiotic recovered mucosa and muscular layers' thickness to C levels (p<0.05). I-FABP and L-FABP expressions in NECP showed intermediate values between C and NEC. CONCLUSION: L. acidophilus had a protective effect on the development of NEC and FABPs could demonstrate the degree of tissular damage of the intestine.
Assuntos
Enterocolite Necrosante/metabolismo , Proteínas de Ligação a Ácido Graxo/biossíntese , Lactobacillus acidophilus , Probióticos/uso terapêutico , Animais , Animais Recém-Nascidos , Biomarcadores/metabolismo , Western Blotting , Modelos Animais de Doenças , Enterocolite Necrosante/terapia , Feminino , Humanos , Imuno-Histoquímica , Lactente , Recém-Nascido , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
The congenital diaphragmatic hernia is a defect in the formation of the diaphragm, which affects between 1:2,000 and 1:4,000 live births and represents 8% of major congenital anomalies. Medical advances in the last 30 years involving prenatal diagnosis, fetal intervention, neonatal surgical and clinical management have changed the survival of these patients. The historical evolution of these advances helps us to understand the effort in pursuit of better results of this defect, which is often lethal. Perspectives on the use of bioengineering and therapy involving stem cells may bring new hope for fetuses with congenital diaphragmatic hernia.
Assuntos
Hérnias Diafragmáticas Congênitas/diagnóstico , Hérnias Diafragmáticas Congênitas/terapia , Diagnóstico Pré-Natal , Feminino , Feto/cirurgia , Previsões , Humanos , Gravidez , Diagnóstico Pré-Natal/tendênciasRESUMO
BACKGROUND/PURPOSE: Congenital diaphragmatic hernia (CDH) is a defect that presents high mortality because of pulmonary hypoplasia and hypertension. Mechanical ventilation changes signaling pathways, such as nitric oxide and VEGF in the pulmonary arterioles. We investigated the production of NOS2 and NOS3 and expression of VEGF and its receptors after ventilation in rat fetuses with CDH. METHODS: CDH was induced by Nitrofen. The fetuses were divided into 6 groups: 1) control (C); 2) control ventilated (CV); 3) exposed to nitrofen (N-); 4) exposed to nitrofen ventilated (N-V), 5) CDH and 6) CDH ventilated (CDHV). Fetuses were harvested and ventilated. We assessed body weight (BW), total lung weight (TLW), TLW/BW ratio, the median pulmonary arteriolar wall thickness (MWT). We analyzed the expression of NOS2, NOS3, VEGF and its receptors by immunohistochemistry and Western blotting. RESULTS: BW, TLW, and TLW/BW ratio were greater on C than on N- and CDH (p<0.05). The MWT was higher in CDH than in CDHV (p<0.001). CDHV showed increased expression of NOS3 (p<0.05) and VEGFR1 (p<0.05), but decreased expression of NOS2 (p<0.05) and VEGFR2 (p<0.001) compared to CDH. CONCLUSION: Ventilation caused pulmonary vasodilation and changed the expression of NOS and VEGF receptors.
Assuntos
Hérnia Diafragmática/metabolismo , Hérnias Diafragmáticas Congênitas/metabolismo , Óxido Nítrico Sintase/metabolismo , Respiração Artificial , Fator A de Crescimento do Endotélio Vascular/metabolismo , Vasodilatação/fisiologia , Animais , Animais Recém-Nascidos , Modelos Animais de Doenças , Feminino , Hérnia Diafragmática/induzido quimicamente , Hérnia Diafragmática/fisiopatologia , Ratos , Ratos Sprague-DawleyRESUMO
A hérnia diafragmática congênita é um defeito de formação do diafragma que acomete entre 1:2.000 e 1:4.000 nascidos vivos e constitui 8% das principais anomalias congênitas. Avanços médicos nos últimos 30 anos envolvendo diagnóstico pré-natal, intervenção fetal, manejo clinico e cirúrgico neonatal têm mudado a sobrevivência dos seus portadores. A evolução histórica desses avanços ajuda a compreender o esforço na busca de melhores resultados desse defeito muitas vezes fatal. Perspectivas na utilização de bioengenharia e terapia envolvendo células tronco podem trazer novas esperanças para os fetos com hérnia diafragmática congênita.
The congenital diaphragmatic hernia is a defect in the formation of the diaphragm, which affects between 1:2,000 and 1:4,000 live births and represents 8% of major congenital anomalies. Medical advances in the last 30 years involving prenatal diagnosis, fetal intervention, neonatal surgical and clinical management have changed the survival of these patients. The historical evolution of these advances helps us to understand the effort in pursuit of better results of this defect, which is often lethal. Perspectives on the use of bioengineering and therapy involving stem cells may bring new hope for fetuses with congenital diaphragmatic hernia.
Assuntos
Humanos , Feminino , Gravidez , Hérnias Diafragmáticas Congênitas/diagnóstico , Hérnias Diafragmáticas Congênitas/terapia , Diagnóstico Pré-Natal , Feto/cirurgia , Previsões , Diagnóstico Pré-Natal/tendênciasRESUMO
PURPOSE: The use of dexamethasone (Dx) stimulates growth, fetal lung maturation and can improve pulmonary hypertension in congenital diaphragmatic hernia (CDH). Our aim was to evaluate the effect of Dx on the lung after fetal pulmonary ventilation in the CDH rat model. METHODS: Some groups underwent prenatal treatment with dexamethasone (0.4 mg/kg) that was given at 18.5 gestational day (GD). Sprague-Dawley rat fetuses were divided into groups: control (C); ventilated control (CV); control exposed to dexamethasone (CDx); ventilated control exposed to dexamethasone (CVDx); congenital diaphragmatic hernia (CDH), ventilated CDH (CDHV), CDH exposed to dexamethasone (CDHDx) and ventilated CDH exposed to dexamethasone (CDHVDx). At 21.5 GD fetuses were delivered by C-section, weighed and ventilated for 30 min. We analyzed the lung morphometry by Masson's Trichrome stain, and VEGF, VEGFR1, VEGFR2 and NOS3 expression by immunohistochemistry. RESULTS: All fetuses with CDH, with or without prenatal dexamethasone showed lung and body weight lower than control fetuses (p < 0.05). All groups that received dexamethasone showed a decrease in the medial muscular layer of arterioles, the internal diameter of the air spaces (Lma) and length of parenchymal transection/airspace ratio (p < 0.05). In the immunohistochemistry, VEGF decreased more in CDHDV group (p < 0.05). VEGFR1 showed no difference, whereas VEGFR2 decreased significantly in the CDHDV group (p < 0.05). NOS3 increased in the group CDHDV (p < 0.05). CONCLUSION: The use of prenatal dexamethasone added to ventilation alters the VEGF and NO pathways.
Assuntos
Dexametasona/uso terapêutico , Hérnias Diafragmáticas Congênitas/prevenção & controle , Pulmão/embriologia , Óxido Nítrico/biossíntese , Prenhez , Respiração Artificial/métodos , Fator A de Crescimento do Endotélio Vascular/biossíntese , Animais , Modelos Animais de Doenças , Feminino , Glucocorticoides/uso terapêutico , Hérnias Diafragmáticas Congênitas/embriologia , Hérnias Diafragmáticas Congênitas/metabolismo , Imuno-Histoquímica , Gravidez , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To standardize a technique for ventilating rat fetuses with Congenital Diaphragmatic Hernia (CDH) using a volume-controlled ventilator. METHODS: Pregnant rats were divided into the following groups: a) control (C); b) exposed to nitrofen with CDH (CDH); and c) exposed to nitrofen without CDH (N-). Fetuses of the three groups were randomly divided into the subgroups ventilated (V) and non-ventilated (N-V). Fetuses were collected on day 21.5 of gestation, weighed and ventilated for 30 minutes using a volume-controlled ventilator. Then the lungs were collected for histological study. We evaluated: body weight (BW), total lung weight (TLW), left lung weight (LLW), ratios TLW / BW and LLW / BW, morphological histology of the airways and causes of failures of ventilation. RESULTS: BW, TLW, LLW, TLW / BW and LLW / BW were higher in C compared with N- (p <0.05) and CDH (p <0.05), but no differences were found between the subgroups V and N-V (p> 0.05). The morphology of the pulmonary airways showed hypoplasia in groups N- and CDH, with no difference between V and N-V (p <0.05). The C and N- groups could be successfully ventilated using a tidal volume of 75 ìl, but the failure of ventilation in the CDH group decreased only when ventilated with 50 ìl. CONCLUSION: Volume ventilation is possible in rats with CDH for a short period and does not alter fetal or lung morphology.
Assuntos
Terapias Fetais/métodos , Hérnias Diafragmáticas Congênitas/terapia , Respiração Artificial/métodos , Respiração Artificial/normas , Animais , Desenho de Equipamento , Feminino , Gravidez , Ventilação Pulmonar , Ratos , Ratos Sprague-Dawley , Respiração Artificial/instrumentaçãoRESUMO
OBJECTIVE: To standardize a technique for ventilating rat fetuses with Congenital Diaphragmatic Hernia (CDH) using a volume-controlled ventilator. METHODS: Pregnant rats were divided into the following groups: a) control (C); b) exposed to nitrofen with CDH (CDH); and c) exposed to nitrofen without CDH (N-). Fetuses of the three groups were randomly divided into the subgroups ventilated (V) and non-ventilated (N-V). Fetuses were collected on day 21.5 of gestation, weighed and ventilated for 30 minutes using a volume-controlled ventilator. Then the lungs were collected for histological study. We evaluated: body weight (BW), total lung weight (TLW), left lung weight (LLW), ratios TLW / BW and LLW / BW, morphological histology of the airways and causes of failures of ventilation. RESULTS: BW, TLW, LLW, TLW / BW and LLW / BW were higher in C compared with N- (p <0.05) and CDH (p <0.05), but no differences were found between the subgroups V and N-V (p> 0.05). The morphology of the pulmonary airways showed hypoplasia in groups N- and CDH, with no difference between V and N-V (p <0.05). The C and N- groups could be successfully ventilated using a tidal volume of 75 ìl, but the failure of ventilation in the CDH group decreased only when ventilated with 50 ìl. CONCLUSION: Volume ventilation is possible in rats with CDH for a short period and does not alter fetal or lung morphology. .
OBJETIVO: padronizar uma técnica para ventilar fetos de rato com HDC usando um ventilador volume-controlado. MÉTODOS: ratas grávidas foram distribuídas em: a) Controle (C); e b) Expostos a Nitrofen com HDC e sem HDC (N-). Fetos dos três grupos foram divididos aleatoriamente em subgrupos ventilados (V) ou não ventilados (NV). Os fetos foram coletados no dia 21,5 da gestação, pesados e ventilados por 30 minutos usando um ventilador volume-controlado. A seguir os pulmões foram coletados para estudo histológico. Nós avaliamos: peso corporal (PC), peso pulmonar total (PPT), peso do pulmão esquerdo (PPE), razão PPT/PC e PPE/PC, histologia morfológica das vias aéreas e as causas das falhas da ventilação. RESULTADOS: PC, PPT, PPE, LLW, PPT/PC e PPE/PC foram maiores em C em relação a N- (p<0,05) e a HDC (p<0,05), mas não houve diferenças entre os subgrupos V e NV (p>0,05). A morfologia das vias aéreas pulmonares mostrou hipoplasia nos grupos N- e HDC, não havendo diferença entre V e NV (p<0,05). Os grupos C e N- puderam ser ventilados com sucesso usando o volume corrente de 75ìl, mas a falha de ventilação no grupo HDC só diminuiu quando ventilados com 50ìl. . CONCLUSÃO: a ventilação a volume de ratos com HDC por um curto período é possível e não altera a morfologia fetal ou pulmonar. .
Assuntos
Animais , Feminino , Gravidez , Ratos , Terapias Fetais/métodos , Hérnias Diafragmáticas Congênitas/terapia , Respiração Artificial/métodos , Respiração Artificial/normas , Desenho de Equipamento , Ventilação Pulmonar , Ratos Sprague-Dawley , Respiração Artificial/instrumentaçãoRESUMO
PURPOSE: To evaluate the expression of myosin in muscle fibers of the diaphragm in experimental congenital diaphragmatic hernia (CDH). METHODS: Fetuses of pregnant rats were divided into four groups: External Control (EC), composed of non-manipulated rats; Nitrofen, composed of pregnant rats that received 100 mg of nitrofen (2,4-dichloro-4'nitrodiphenyl ether) diluted in olive oil on gestational day (GD) 9.5, whose fetuses developed CDH (N+) or not (N-), and Olive Oil Placebo (OO), composed of pregnant rats that received the oil on the same GD. The fetuses were collected on GD 18.5, 19.5, 20.5 and 21.5 (term = 22 days). We obtained body weight (BW) and photographed the diaphragm area (DA), hernia area (HA) and subsequent calculated the HA/DA ratio in N+ group. Samples of Diaphragm muscle were processed for histological staining with H/E and immunohistochemistry (IHQ) for myosin. RESULTS: The fetuses of N- and N+ groups had decreased BW and DA compared to EC and OO groups (p < 0.001). HA was decreased on GD 18.5 compared to 21.5 (p < 0.001) and the HA/DA ratio showed no difference. IHQ showed decreased expression of myosin in nitrofen groups. CONCLUSION: CDH induced by nitrofen model contributes to the understanding of muscularization in the formation of the diaphragm where the myosin expression is decreased.