Abstract
Matrix metalloproteinases (MMP) are likely effectors of normal lung development, especially branching morphogenesis, angiogenesis, and extracellular matrix degradation. Because hyperoxia exposure (>95% O2) from d 4 to 14 in newborn rat pups leads to arrest of alveolarization and mimics newborn chronic lung disease, we tested whether hyperoxia altered MMP-2 and -9 mRNA, protein, and enzymatic activity, and the mRNA and protein expression of the endogenous tissue inhibitor of MMP, TIMP-1. No changes due to hyperoxia exposure were observed in MMP-2 mRNA or pro-enzyme (72 kD) protein levels between d 6 and 14, although the overall protein mass and zymographic activity of the active (68 kD) enzyme were diminished (p < 0.05, ANOVA). However, hyperoxia significantly decreased levels of MMP-9 mRNA and pro-MMP-9 protein and diminished overall MMP-9 pro-enzyme activity. TIMP-1 mRNA was not elevated by hyperoxia until d 14, but protein levels were significantly (p < 0.001) elevated by hyperoxia from d 9 to 14. To estimate the potential of MMP inhibition to arrest alveolarization, administration of doxycycline (20 mg/ kg, twice daily by gavage), a pan-MMP proteolysis inhibitor, arrested lung alveolarization. We conclude that hyperoxia decreases MMP-9 mRNA, protein, and activity and elevates TIMP-1 protein, and these changes have the potential to contribute to the arrest of normal lung development.
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Abbreviations
- MMP:
-
matrix metalloproteinase
- TIMP:
-
tissue inhibitor of matrix metalloproteinase
- RT:
-
reverse transcription
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The authors thank Ms. N. Hooton and Mr. D. Zaragoza for technical assistance, and Ms. Christina Maley and Ms. Sheila McManus for editorial assistance.
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Supported by the Canadian Institutes of Health Research, the Alberta Heritage Foundation for Medical Research, the Alberta Lung Association, and the Hospital for Sick Children Foundation. G.E.H. was funded by a studentship from the Alberta Heritage Foundation for Medical Research.
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Hosford, G., Fang, X. & Olson, D. Hyperoxia Decreases Matrix Metalloproteinase-9 and Increases Tissue Inhibitor of Matrix Metalloproteinase-1 Protein in the Newborn Rat Lung: Association with Arrested Alveolarization. Pediatr Res 56, 26–34 (2004). https://doi.org/10.1203/01.PDR.0000130658.45564.1F
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DOI: https://doi.org/10.1203/01.PDR.0000130658.45564.1F
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