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Respiratory Research
Published in: Respiratory Research 1/2006

Open Access 01-12-2006 | Research

Apoptosis and the activity of ceramide, Bax and Bcl-2 in the lungs of neonatal rats exposed to limited and prolonged hyperoxia

Authors: Ahmad W Husari, Ghassan S Dbaibo, Hala Bitar, Aline Khayat, Shoghag Panjarian, Michel Nasser, Fadi F Bitar, Marwan El-Sabban, Ghazi Zaatari, Salman M Mroueh

Published in: Respiratory Research | Issue 1/2006

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Abstract

Background

The aim of the study is to examine the effect of limited and prolonged hyperoxia on neonatal rat lung. This is done by examining the morphologic changes of apoptosis, the expression of ceramide, an important mediator of apoptosis, the expression of inflammatory mediators represented by IL-1β and the expression of 2 proto-oncogenes that appear to modulate apoptosis (Bax and Bcl-2).

Methods

Newborn rats were placed in chambers containing room air or oxygen above 90% for 7 days. The rats were sacrificed at 3, 7 or 14 days and their lungs removed. Sections were fixed, subjected to TUNEL, Hoechst, and E-Cadherin Staining. Sections were also incubated with anti-Bcl-2 and anti-Bax antisera. Bcl-2 and Bax were quantitated by immunohistochemistry. Lipids were extracted, and ceramide measured through a modified diacylglycerol kinase assay. RT-PCR was utilized to assess IL-1β expression.

Results

TUNEL staining showed significant apoptosis in the hyperoxia-exposed lungs at 3 days only. Co-staining of the apoptotic cells with Hoechst, and E-Cadherin indicated that apoptotic cells were mainly epithelial cells. The expression of Bax and ceramide was significantly higher in the hyperoxia-exposed lungs at 3 and 14 days of age, but not at 7 days. Bcl-2 was significantly elevated in the hyperoxia-exposed lungs at 3 and 14 days. IL-1β expression was significantly increased at 14 days.

Conclusion

Exposure of neonatal rat lung to hyperoxia results in early apoptosis documented by TUNEL assay. The early rise in Bax and ceramide appears to overcome the anti-apoptotic activity of Bcl-2. Further exposure did not result in late apoptotic changes. This suggests that apoptotic response to hyperoxia is time sensitive. Prolonged hyperoxia results in acute lung injury and the shifting balance of ceramide, Bax and Bcl-2 may be related to the evolution of the inflammatory process.
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Metadata
Title
Apoptosis and the activity of ceramide, Bax and Bcl-2 in the lungs of neonatal rats exposed to limited and prolonged hyperoxia
Authors
Ahmad W Husari
Ghassan S Dbaibo
Hala Bitar
Aline Khayat
Shoghag Panjarian
Michel Nasser
Fadi F Bitar
Marwan El-Sabban
Ghazi Zaatari
Salman M Mroueh
Publication date
01-12-2006
Publisher
BioMed Central
Published in
Respiratory Research / Issue 1/2006
Electronic ISSN: 1465-993X
DOI
https://doi.org/10.1186/1465-9921-7-100

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