Gene expression profile in newborn rat lungs after two days of recovery of mechanical ventilation.
BACKGROUND
Preterm infants having immature lungs often require respiratory support, potentially leading to bronchopulmonary dysplasia (BPD). Conventional BPD rodent models based on mechanical ventilation (MV) present outcome measured at the end of the ventilation period. A reversible intubation and ventilation model in newborn rats recently allowed discovering that different sets of genes modified their expression related to time after MV. In a newborn rat model, the expression profile 48 h after MV was analyzed with gene arrays to detect potentially interesting candidates with an impact on BPD development.


METHODS
Rat pups were injected P4-5 with 2 mg/kg lipopolysaccharide (LPS). One day later, MV with 21 or 60% oxygen was applied during 6 h. Animals were sacrified 48 h after end of ventilation. Affymetrix gene arrays assessed the total gene expression profile in lung tissue.


RESULTS
In fully treated animals (LPS + MV + 60% O(2)) vs. controls, 271 genes changed expression significantly. All modified genes could be classified in six pathways: tissue remodeling/wound repair, immune system and inflammatory response, hematopoiesis, vasodilatation, and oxidative stress. Major alterations were found in the MMP and complement system.


CONCLUSIONS
MMPs and complement factors play a central role in several of the pathways identified and may represent interesting targets for BPD treatment/prevention.Bronchopulmonary dysplasia (BPD) is a chronic lung disease occurring in ~30% of preterm infants born less than 30 wk of gestation (1). Its main risk factors include lung immaturity due to preterm delivery, mechanical ventilation (MV), oxygen toxicity, chorioamnionitis, and sepsis. The main feature is an arrest of alveolar and capillary formation (2). Models trying to decipher genes involved in the pathophysiology of BPD are mainly based on MV and oxygen application to young mammals with immature lungs of different species (3). In newborn rodent models, analyses of lung structure and gene and protein expression are performed for practical reasons directly at the end of MV (4,5,6). However, later appearing changes of gene expression might also have an impact on lung development and the evolution towards BPD and cannot be discovered by such models. Recently, we developed a newborn rat model of MV using an atraumatic (orotracheal) intubation technique that allows the weaning of the newborn animal off anesthesia and MV, the extubation to spontaneous breathing, and therefore allows the evaluation of effects of MV after a ventilation-free period of recovery (7). Indeed, applying this concept of atraumatic intubation by direct laryngoscopy, we recently were able to show significant differences between gene expression changes appearing directly after MV compared to those measured after a ventilation-free interval of 48 h. Immediately after MV, inflammation-related genes showed a transitory modified expression, while another set of more structurally related genes changed their expression only after a delay of 2 d (7). Lung structure, analyzed by conventional 2D histology and also by 3D reconstruction using synchrotron x-ray tomographic microscopy revealed, 48 h after end of MV, a reduced complexity of lung architecture compared to the nonventilated rat lungs, similar to the typical findings in BPD. To extend these observations about late gene expression modifications, we performed with a similar model a full gene expression profile of lung tissue 48 h after the end of MV with either room air or 60% oxygen. Essentially, we measured changes in the expression of genes related to the MMPs and complement system which played a role in many of the six identified mostly affected pathways.
Department of Pediatrics, Clinic of Neonatology, University Hospital of Lausanne, Lausanne, Switzerland.
Valérie
V
Dénervaud
Department of Pediatrics, Clinic of Neonatology, University Hospital of Lausanne, Lausanne, Switzerland.
Sandrine
S
Gremlich
Department of Pediatrics, Clinic of Neonatology, University Hospital of Lausanne, Lausanne, Switzerland.
Eliane
E
Trummer-Menzi
Institute of Anatomy, University of Bern, Bern, Switzerland.
Johannes C
JC
Schittny
Department of Pediatrics, Clinic of Neonatology, University Hospital of Lausanne, Lausanne, Switzerland.
Matthias
M
Roth-Kleiner
D000818:Animals; D000831:Animals, Newborn; D001997:Bronchopulmonary Dysplasia / Q000235:genetics*; D004195:Disease Models, Animal; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D008070:Lipopolysaccharides; D008168:Lung / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D008171:Lung Diseases / Q000139:chemically induced / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D017208:Rats, Wistar; D012121:Respiration, Artificial / Q000009:adverse effects*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors
D008070:Lipopolysaccharides; C433788:lipopolysaccharide, E. coli O26-B6