title: Time course of housekeeping gene expression changes in diffuse alveolar damage induced by hyperoxia exposure in mice.
abstract: We have found diffuse alveolar damage (DAD) has taken place in some patients under mechanical ventilation with high-inspired oxygen concentrations. To clarify the molecular pathophysiology of this, the time course of gene expression changes induced by hyperoxia exposure in mouse lungs was examined using real-time quantitative polymerase chain reaction (real-time qPCR). Our raw data and those normalized with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) showed that: (1) there is a decrease in levels of mRNAs for surfactant-associated protein C (SFTPC), cytochrome P450, 2F2 (CYP2F2), Claudin 1 (CLDN1), membrane-associated zonula occludens protein-1 (ZO-1), lysozyme (LYZS), and this suggests alveolar dysfunction and a disruption of the immune system, (2) we confirmed apoptotic conditions, such as significant up-regulations of mRNA levels in Myc and Galectin-3, and (3) hyperoxic conditions probably yielded reactive oxygen species (ROS), which resulted in a malignant cycle of ROS production by Myc overexpression [Shimada I, Matsui K, Brinkmann B, Hohoff C, Hiraga K, Tabuchi Y, et al. Novel transcript profiling of diffuse alveolar damage induced by hyperoxia exposure in mice: normalization by glyceraldehyde 3-phosphate dehydrogenase. Int J Legal Med 2008;122:373-83]. In this experiment, GAPDH was up-regulated when hyperoxia exposure was continued. Therefore, we reexamined our data and found that: (1) mRNA levels of other housekeeping genes, including beta(2)-microglobulin (beta2M), ribosomal protein: large P2 (RPLP2), and importin 8 (IPO8) altered to a lesser extent, (2) mRNA levels of beta2M and IPO8 were down-regulated when hyperoxia exposure was continued, and (3) our previous work was validated by normalization with these three housekeeping genes.
authors.affiliation: Department of Forensic Medicine, Faculty of Medical Sciences, University of Fukui, 23-3, Matsuokashimoaizuki, eiheiji-cho, Yoshida-gun, Fukui 910-1193, Japan. ichi@u-fukui.ac.jp
authors.forename: Ichiroh
authors.initials: I
authors.lastname: Shimada
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Matsui
authors.forename: Reiko
authors.initials: R
authors.lastname: Iida
authors.forename: Etsuko
authors.initials: E
authors.lastname: Tsubota
authors.forename: Takasumi
authors.initials: T
authors.lastname: Matsuki
mesh_terms: D048948:14-3-3 Proteins; D000818:Animals; D015536:Down-Regulation; D015870:Gene Expression; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics / Q000378:metabolism; D018496:Hyperoxia / Q000235:genetics* / Q000473:pathology*; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D010735:Phosphoglycerate Kinase / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D011650:Pulmonary Alveoli / Q000473:pathology*; D037721:Pulmonary Surfactant-Associated Protein C; D012333:RNA, Messenger / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D011990:Receptors, Transferrin / Q000235:genetics / Q000378:metabolism; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D025822:Ubiquitin C / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation; D001613:beta 2-Microglobulin / Q000235:genetics / Q000378:metabolism; D028961:beta Karyopherins / Q000235:genetics / Q000378:metabolism
chemical_list: D048948:14-3-3 Proteins; C107282:IPO8 protein, human; D036341:Intercellular Signaling Peptides and Proteins; D018832:Molecular Chaperones; C489428:Myc protein, mouse; D010455:Peptides; D010750:Phosphoproteins; D016271:Proto-Oncogene Proteins c-myc; D037721:Pulmonary Surfactant-Associated Protein C; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D011990:Receptors, Transferrin; D012269:Ribosomal Proteins; C489680:Sftpc protein, mouse; D025822:Ubiquitin C; C490391:Ywhaz protein, rat; D001613:beta 2-Microglobulin; D028961:beta Karyopherins; C031937:phosphoprotein P2, ribosomal; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D007041:Hypoxanthine Phosphoribosyltransferase; C528165:Pgk1 protein, mouse; D010735:Phosphoglycerate Kinase