title: Apoptosis and the activity of ceramide, Bax and Bcl-2 in the lungs of neonatal rats exposed to limited and prolonged hyperoxia.
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-1beta 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-1beta 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-1beta expression was significantly increased at 14 days.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, American University of Beirut-Medical Center, P,O, Box: 113-6044, Beirut 1107 2802, Lebanon. ah51@aub.edu.lb
authors.forename: Ahmad W
authors.initials: AW
authors.lastname: Husari
authors.forename: Ghassan S
authors.initials: GS
authors.lastname: Dbaibo
authors.forename: Hala
authors.initials: H
authors.lastname: Bitar
authors.forename: Aline
authors.initials: A
authors.lastname: Khayat
authors.forename: Shoghag
authors.initials: S
authors.lastname: Panjarian
authors.forename: Michel
authors.initials: M
authors.lastname: Nasser
authors.forename: Fadi F
authors.initials: FF
authors.lastname: Bitar
authors.forename: Marwan
authors.initials: M
authors.lastname: El-Sabban
authors.forename: Ghazi
authors.initials: G
authors.lastname: Zaatari
authors.forename: Salman M
authors.initials: SM
authors.lastname: Mroueh
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000502:physiology*; D002518:Ceramides / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D018496:Hyperoxia / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D008168:Lung / Q000254:growth & development / Q000378:metabolism* / Q000473:pathology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D013997:Time Factors; D051028:bcl-2-Associated X Protein / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*
chemical_list: D002518:Ceramides; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein