title: Integrin-mediated suppression of endotoxin-induced DNA damage in lung endothelial cells is sensitive to poly(ADP-ribose) polymerase-1 gene deletion.
abstract: Endotoxin (LPS) is a cause of adult respiratory distress syndrome (ARDS), a disease which is preceded by acute lung injury involving the pulmonary endothelium. Experimentally, LPS causes acute DNA strand breakage in mouse lung endothelial cells (MLEC). Engagement of integrin cell adhesion receptors inhibits acute DNA breakage, which could be of use in reducing lung injury. Because integrins presumably inhibit DNA damage or activate repair, we hypothesized that the DNA-damage response protein, poly(ADP-ribose) polymerase-1 (PARP-1), regulates the protective action of integrins, as well as sensitivity to LPS. Therefore, the effect of LPS on MLEC cultured from wild-type and PARP-1 knockout mice was determined. Fluorescence microscopic measures were used to assess plasma membrane integrity, PARP activity, DNA strand breakage and DNA repair in attached cells. LPS caused a concentration-dependent increase in the permeability of wild-type MLEC. Engagement of beta1 integrins with an antibody protected wild-type MLEC from this LPS-induced injury. Wild-type cells treated with the PARP-inhibitor, 3-aminobenzamide, and PARP-1 knockout MLEC were also resistant. LPS caused acute DNA breakage in both wild-type and knockout MLEC, but PARP was activated only in wild-type cells. LPS-induced DNA breakage was inhibited by 3-aminobenzamide, but not by knockout. Anti-beta1 integrin antibody also inhibited the DNA breakage and PARP activation caused by LPS in wild-type MLEC. However, integrin engagement did not prevent DNA breakage in PARP-1 knockout cells, despite a similar level of beta1 integrin in wild-type and knockout cells. Thus, integrin engagement, 3-aminobenzamide, and PARP-1 deletion protected MLEC from increases in membrane permeability caused by LPS. PARP-1 deletion also impaired the ability of integrin engagement to inhibit LPS-induced DNA breakage, suggesting that knockout may affect nuclear factors necessary for integrin-mediated suppression of LPS-induced DNA breakage.
authors.affiliation: Division of Pharmacology, Ohio State University College of Pharmacy, 500 West Twelfth Avenue, Columbus, OH 43210, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Huang
authors.forename: Jane L
authors.initials: JL
authors.lastname: McIntosh
authors.forename: Lanyan
authors.initials: L
authors.lastname: Fang
authors.forename: Csaba
authors.initials: C
authors.lastname: Szabo
authors.forename: Dale G
authors.initials: DG
authors.lastname: Hoyt
mesh_terms: D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D015153:Blotting, Western; D020169:Caspases / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D004249:DNA Damage; D004260:DNA Repair; D004305:Dose-Response Relationship, Drug; D004727:Endothelium / Q000378:metabolism*; D004731:Endotoxins / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D017353:Gene Deletion; D016023:Integrins / Q000378:metabolism*; D007700:Kinetics; D008070:Lipopolysaccharides / Q000737:chemistry; D008168:Lung / Q000166:cytology* / Q000473:pathology; D051379:Mice; D018345:Mice, Knockout; D008856:Microscopy, Fluorescence; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics*; D013997:Time Factors
chemical_list: D001549:Benzamides; D004731:Endotoxins; D004791:Enzyme Inhibitors; D016023:Integrins; D008070:Lipopolysaccharides; C025160:3-aminobenzamide; D011065:Poly(ADP-ribose) Polymerases; D020169:Caspases