Abstract
Aim:
To test the hypothesis that PI3K/Akt/eNOS signaling has a protective role in a murine model of ventilation associated lung injury (VALI) through down-regulation of p38 MAPK signaling.
Methods:
Male C57BL/J6 (wild-type, WT) or eNOS knockout mice (eNOS−/−) were exposed to mechanical ventilation (MV) with low (LVT, 7 mL/kg) and high tidal volume (HVT, 20 mL/kg) for 0−4 h. A subset of WT mice was administered the specific inhibitors of PI3K (100 nmol/L Wortmannin [Wort], ip) or of p38 MAPK (SB203580, 2 mg/kg, ip) 1 h before MV. Cultured type II alveolar epithelial cells C10 were exposed to 18% cyclic stretch for 2 h with or without 20 nmol/L Wort pretreatment. At the end of the experiment, the capillary leakage in vivo was assessed by extravasation of Evans blue dye (EBD), wet/dry weight ratio and lung lavage protein concentration. The lung tissue and cell lysate were also collected for protein and histological review.
Results:
MV decreased PI3K/Akt phosphorylation and eNOS expression but increased phospho-p38 MAPK expression along with a lung leakage of EBD. Inhibitions of phospho-Akt by Wort worsen the lung edema, whereas inhibition of p38 MAPK kinase restored activation of Akt together with alleviated capillary leakage. eNOS−/− mice showed an exacerbated lung edema and injury. The stretched C10 cells demonstrated that Wort diminished the activation of Akt, but potentiated phosphorylation of MAPK p38.
Conclusion:
Our results indicate that PI-3K/Akt/eNOS pathway has significant protective effects in VALI by preventing capillary leakage, and that there is a cross-talk between PI3K/Akt and p38 MAPK pathways in vascular barrier dysfunction resulting from VALI.
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Acknowledgements
This work was supported by grants from American Heart Association (Mid-Atlantic, Beginning Grant-in-Aid #0765286U), American Lung Association of Maryland (Biomedical Research Grant, 2006) and the National Heart, Lung and Blood Institute (NIH R01 HL049441; P50 HL 73994).
The authors thank Drs Hunter C CHAMPION, Allan CHESLEY and Michael T CROW for helpful consultation and technical assistance. We thank CPT Gleeson MURPHY and Ms Cindy A KRONMAN for critical reading.
Finally, I give heartfelt love and gratitude to my father, Mr Yun-xiang PENG, for his unconditional love and support on my effort to get funding for this work.
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Peng, Xq., Damarla, M., Skirball, J. et al. Protective role of PI3-kinase/Akt/eNOS signaling in mechanical stress through inhibition of p38 mitogen-activated protein kinase in mouse lung. Acta Pharmacol Sin 31, 175–183 (2010). https://doi.org/10.1038/aps.2009.190
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DOI: https://doi.org/10.1038/aps.2009.190
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