Detection of mononuclear cells as the source of the increased tissue factor mRNA in the liver from lipopolysaccharide-treated rats.
Tissue factor (TF) triggers the coagulation cascade reaction in vivo. Overexpression of TF mRNA is one leading cause of disseminated intravascular coagulation and thrombosis-related organ failure. In response to lipopolysaccharide (LPS) stimulation, various cell types can produce TF mRNA in vitro. However, there is currently no agreement on what types of cells in the liver overexpress TF mRNA after LPS treatment. For the first report, we found the increased TF mRNA with reverse transcription-polymerase chain reaction (RT-PCR), and confirmed a fourfold increase (p<0.001 vs. control, t-test) of the TF mRNA level with RT-competitive PCR in the liver of LPS-treated (2.0 mg/kg i.v. injection) rats. There was no significant difference in the glyceraldehyde-3-phosphate dehydrogenase mRNA level between LPS-treated rats and control rats. To clarify the localization and cellular source of LPS-induced TF mRNA, we performed in situ hybridization analysis with [35S]-labeled oligonucleotides probes, which we originally designed. We detected intense signals of TF mRNA in mononuclear cells but not in endothelial cells around the hepatic vein of LPS-treated rats. In this study, we showed that the TF mRNA level induced by LPS treatment, which may indicate mononuclear cells associated, significantly increased in the liver of rats. These results will provide circumstantial support for the therapeutic strategy that mononuclear cell should be one of the target cells to be treated in the early phase of disseminated intravascular coagulation in the liver, and that the need to suppress its overexpression of TF mRNA is essential for preventing hypercoagulable condition.
Arai
A
A
Department of Pediatrics, School of Medicine, University of Occupational and Environmental Health, Yahatanishi, Kitakyushu, Japan.
Hirano
H
H
Ueta
Y
Y
Hamada
T
T
Mita
T
T
Shirahata
A
A
D000818:Animals; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D005260:Female; D006503:Hepatic Veins / Q000166:cytology / Q000378:metabolism; D017403:In Situ Hybridization; D007963:Leukocytes, Mononuclear / Q000737:chemistry*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008099:Liver / Q000166:cytology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000187:drug effects*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013925:Thromboplastin / Q000235:genetics*; D014018:Tissue Distribution
D008070:Lipopolysaccharides; D012333:RNA, Messenger; D013925:Thromboplastin