title: Collagen is a survival factor against LPS-induced apoptosis in cultured sheep pulmonary artery endothelial cells.
abstract: Lipopolysaccharide (LPS) causes direct pulmonary endothelial injury that can precipitate cell death. We investigated the ability of LPS to produce apoptosis in sheep pulmonary artery endothelial cells (SPAEC) grown in monolayer on plastic or collagen. When SPAEC were grown on plastic, LPS (100 ng/ml) caused internucleosomal DNA fragmentation (IDF) to 180- to 200-base pair ladders after 4 h. Higher-order chromatin damage, producing 50-kilobase DNA fragments, occurred within 2 h. Significant DNA strand breaks were seen in attached cells within 1 h incubation with > or = 1 ng LPS/ml, using in situ labeling by break extension (ISBE). DNA strand breakage in attached cells peaked after 2 h and remained elevated after 4 h. Detachment of SPAEC from the monolayer did not begin until 4 h. SPAEC cultured on collagen were protected from LPS-induced apoptosis; DNA damage measured by IDF, high-molecular-weight DNA fragmentation, and ISBE were suppressed. The protective effect of collagen was not due to inactivation of LPS. Thus LPS-induced apoptosis occurs in SPAEC after genotoxic damage and this process is suppressed by the extracellular matrix.
authors.affiliation: Department of Pharmacology, University of Pittsburgh School of Medicine, Pennsylvania 15261, USA.
authors.forename: D G
authors.initials: DG
authors.lastname: Hoyt
authors.forename: R J
authors.initials: RJ
authors.lastname: Mannix
authors.forename: J M
authors.initials: JM
authors.lastname: Rusnak
authors.forename: B R
authors.initials: BR
authors.lastname: Pitt
authors.forename: J S
authors.initials: JS
authors.lastname: Lazo
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology*; D004249:DNA Damage; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D005109:Extracellular Matrix / Q000502:physiology; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008970:Molecular Weight; D009707:Nucleosomes / Q000378:metabolism; D011651:Pulmonary Artery / Q000166:cytology / Q000187:drug effects* / Q000502:physiology
chemical_list: D008070:Lipopolysaccharides; D009707:Nucleosomes; D003094:Collagen