Hyperoxia Disrupts Extracellular Signal-Regulated Kinases 1/2-Induced Angiogenesis in the Developing Lungs
Abstract
:1. Introduction
2. Results
2.1. Hyperoxia Exposure Transiently Activates ERK1/2 in Neonatal Mouse Lungs
2.2. Hyperoxia Exposure Transiently Activates ERK1/2 in Neonatal Mouse Lung Endothelial Cells
2.3. Prolonged Hyperoxia Exposure Interrupts Pulmonary Vascularization in Neonatal Mice
2.4. Hyperoxia Exposure Increases Pulmonary Endothelial Cell Apoptosis in Neonatal Mice
2.5. Hyperoxia Exposure Activates ERK1/2 in HPAECs
2.6. PD98059 Efficiently Inhibits ERK1/2 Activation in HPAECs
2.7. ERK1/2 Inhibition Decreases HPAEC Migration
2.8. ERK1/2 Inhibition Decreases HPAEC Proliferation
2.9. ERK1/2 Inhibition Decreases HPAEC Tubule and Mesh Formation
2.10. ERK1/2 Inhibition Alters the Level of Proteins That Regulate Cell Cycle Progression in HPAECs
3. Discussion
4. Materials and Methods
4.1. In Vivo Experiments
4.1.1. Animals
4.1.2. Hyperoxia Experiments
4.1.3. Lung Tissue Harvest and Protein Extraction
4.1.4. Western Blot Assays
4.1.5. Tissue Preparation for Immunofluorescence and Lung Vascular Morphometry Studies
4.1.6. Immunofluorescence Studies
4.1.7. Analyses of Pulmonary Vascularization
4.2. In Vitro Experiments
4.2.1. Cell Culture
4.2.2. Hyperoxia Exposure
4.2.3. Cell Treatment
4.2.4. Western Blot Assays
4.2.5. Cell Proliferation Assay
4.2.6. Scratch Assay
4.2.7. Tubule and Mesh Formation Assays
4.3. Statistical Analyses
Author Contributions
Acknowledgments
Conflicts of Interest
Abbreviations
BPD | bronchopulmonary dysplasia |
Cdk | cyclin-dependent kinase |
ERK | extracellular signal-regulated kinases |
DAPI | 4′,6-diamidino-2-phenylindole |
HPAECs | human pulmonary artery endothelial cells |
MAP kinase | mitogen-activated protein kinase |
MTT | 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide |
PD | PD98059 |
PH | pulmonary hypertension |
NO | nitric oxide |
PND | postnatal day |
WT | wild type |
VEGF | vascular endothelial growth factor |
vWF | von Willebrand factor |
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Menon, R.T.; Shrestha, A.K.; Barrios, R.; Shivanna, B. Hyperoxia Disrupts Extracellular Signal-Regulated Kinases 1/2-Induced Angiogenesis in the Developing Lungs. Int. J. Mol. Sci. 2018, 19, 1525. https://doi.org/10.3390/ijms19051525
Menon RT, Shrestha AK, Barrios R, Shivanna B. Hyperoxia Disrupts Extracellular Signal-Regulated Kinases 1/2-Induced Angiogenesis in the Developing Lungs. International Journal of Molecular Sciences. 2018; 19(5):1525. https://doi.org/10.3390/ijms19051525
Chicago/Turabian StyleMenon, Renuka T., Amrit Kumar Shrestha, Roberto Barrios, and Binoy Shivanna. 2018. "Hyperoxia Disrupts Extracellular Signal-Regulated Kinases 1/2-Induced Angiogenesis in the Developing Lungs" International Journal of Molecular Sciences 19, no. 5: 1525. https://doi.org/10.3390/ijms19051525