title: Endotoxin-induced activation of the nuclear transcription factor kappa B and expression of E-selectin messenger RNA in hepatocytes, Kupffer cells, and endothelial cells in vivo.
abstract: Endotoxin causes neutrophil sequestration in the liver and severe vascular and parenchymal cell injury. Inducible adhesion molecules such as intercellular adhesion molecule-1 and selectins are involved in neutrophil recruitment to an inflammatory site in vivo. The objective of our study was to characterize the translocation of the nuclear factor kappa B (NF-kappa B) from the cytoplasm to the nucleus (NF-kappa B activation) during endotoxemia using the electrophoretic mobility shift assay and to investigate its role in regulation of E-selectin gene transcription in liver cells in vivo. Administration of 0.5 mg/kg Salmonella enteritidis endotoxin to male Fischer rats induced a drastic but transient activation of NF-kappa B at the whole liver level. Major subunits identified were p50 (NF-kappa B1), p65 (RelA), and c-Rel, but not p52 (NF-kappa B2). Isolation of liver cells from control animals induced substantial NF-kappa B activation in Kupffer cells and endothelial cells and minor activation in hepatocytes. One hour after endotoxin treatment in vivo, NF-kappa B was uniformly activated in all isolated cells. At 5 h, NF-kappa B activation was reduced to various degrees in all cell types, with hepatocytes showing only a moderate decrease. Northern blot analysis indicated no E-selectin mRNA in all control cells; however, at 1 h, endotoxin induced E-selectin gene transcription transiently in endothelial cells and in Kupffer cells but not in hepatocytes. These data support the hypothesis that NF-kappa B activation is important for E-selectin gene transcription in hepatic vascular lining cells. However, the fact that hepatic parenchymal cells, despite NF-kappa B activation do not express E- selectin mRNA, suggests that NF-kappa B activation alone is not sufficient for E-selectin gene transcription in vivo.
authors.affiliation: Cardivoscular Pharmacology, Pharmacia and Upjohn, Inc., Kalamazoo, MI 49007, USA.
authors.forename: N A
authors.initials: NA
authors.lastname: Essani
authors.forename: G M
authors.initials: GM
authors.lastname: McGuire
authors.forename: A M
authors.initials: AM
authors.lastname: Manning
authors.forename: H
authors.initials: H
authors.lastname: Jaeschke
mesh_terms: D000818:Animals; D001483:Base Sequence; D019040:E-Selectin / Q000235:genetics*; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D004731:Endotoxins / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology*; D007728:Kupffer Cells / Q000276:immunology / Q000378:metabolism*; D008099:Liver / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D011916:Rats, Inbred F344
chemical_list: D019040:E-Selectin; D004731:Endotoxins; D016328:NF-kappa B; D012333:RNA, Messenger