Gene expression profiling of target genes in ventilator-induced lung injury.
In the lungs, high-pressure mechanical ventilation induces an inflammatory response similar to that observed in acute respiratory distress syndrome. To further characterize these responses and to compare them with classical inflammatory pathways, we performed gene expression profiling analysis of 20,000 mouse genes in isolated blood-free (to exclude genes from sequestered leukocytes) perfused mouse lungs exposed to low-pressure ventilation (10 cmH2O), high-pressure ventilation (25 cmH2O, overventilation), and LPS treatment. A large number of inflammatory and apoptotic genes were increased by both overventilation and LPS. However, certain growth factor-related genes, as well as genes related to development, cellular communication, and the cytoskeleton, were only regulated by overventilation. We validated and confirmed increased mRNA expression pattern of five genes (amphiregulin, gravin, Nur77, Cyr61, interleukin-11) by real-time PCR; furthermore, we confirmed increased protein expression of amphiregulin by immunohistochemistry and immunoblotting assays. These genes represent novel candidate genes in ventilator-induced lung injury.
Dolinay
Tamás
T
Division of Pulmonary, Allergy and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15213, USA.
Kaminski
Naftali
N
Felgendreher
Martina
M
Kim
Hong P
HP
Reynolds
Paul
P
Watkins
Simon C
SC
Karp
Dörte
D
Uhlig
Stefan
S
Choi
Augustine M K
AM
D054758:A Kinase Anchor Proteins; D066258:Amphiregulin; D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D016000:Cluster Analysis; D055515:Cysteine-Rich Protein 61; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D066255:EGF Family of Proteins; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D017370:Interleukin-11 / Q000235:genetics / Q000378:metabolism; D008070:Lipopolysaccharides; D008168:Lung / Q000187:drug effects / Q000378:metabolism; D055370:Lung Injury; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism; D011987:Receptors, Steroid / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012121:Respiration, Artificial / Q000009:adverse effects*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D054758:A Kinase Anchor Proteins; C491161:Akap12 protein, mouse; D066258:Amphiregulin; C587849:Areg protein, mouse; C491850:CCN1 protein, mouse; D018797:Cell Cycle Proteins; D055515:Cysteine-Rich Protein 61; D004268:DNA-Binding Proteins; D066255:EGF Family of Proteins; D006023:Glycoproteins; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D017370:Interleukin-11; D008070:Lipopolysaccharides; C539818:Nr4a1 protein, mouse; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D011987:Receptors, Steroid; D014157:Transcription Factors