Subscribe to RSS
DOI: 10.1055/s-0038-1649850
Plasminogen Activator Inhibitor-1 mRNA Is Induced by Hemorrhage in Endothelial and/Mesothelial Cells of the Rat Liver
Publication History
Received 20 February 1995
Accepted after resubmission 02 May 1995
Publication Date:
09 July 2018 (online)
Summary
We have recently shown that plasminogen activator inhibitor-1 (PAI-I) mRNA is elevated after hemorrhage in various tissues including liver. In this study, we set out to identify the cell types in the liver that are responsible for the increase in PAH mRNA after hemorrhage using in situ reverse transcription-polymerase chain reaction (in situ RT-PCR). Male Sprague-Dawley rats were cannulated and subjected to a 20 ml/kg hemorrhage within 3 min or 300 μg/kg of endotoxin. Pour hours later, the livers were harvested, fixed, frozen, and sectioned. RT-PCR showed an increase of PAI-I mRNA in liver 4 h after hemorrhage or endotoxin-treatment. Standard in situ hybridization could not detect PAI-I mRNA in the livers of either the hemorrhage, endotoxin, or control groups. However, in situ RT-PCR detected PAI-1 mRNA in vascular endothelial cells and capsular mesothelial cells, but not in hepatocytes, in both the hemorrhage and endotoxin groups. No signal was found in the control rats, or when the experimental protocol was modified to 1) omit the RT step, 2) precede the RT step with RNA digestion, or 3) use an irrelevant probe. These results demonstrate that hemorrhage induces PAI-1 mRNA in endothelial and mesothelial cells of liver.
-
References
- 1 Krishnamurti C, Alving BM. Plasminogen activator inhibitor type 1 biochemistry and evidence for modulation of fibrinolysis invivo. Semin Thromb Haemost 1992; 18: 67-80
- 2 Yamashita M, Darlington DN, Weeks EJ, Jones RO, Gann DS. Plasminogen activator inhibitor-1 rises after hemorrhage in the rat. Am J Physiol (Endocrinol Metab) 1995; 268: 1065-1069
- 3 Konkle B, Schuster SJ, Kelly MD, Harjes K, Hassett DE, Bohrer M, Tavas- soli M. Plasminogen activator inhibitor-1 messenger RNA expression is induced in rat hepatocytes in vivo by dexamethasone. Blood 1992; 79: 2636-2642
- 4 Quax PH A, van den Hoogen CM, Verheijen JH, Padro T, Zeheb R, Gelehrter TD, van Berkel TJ C, Kuiper J, Emeis JJ. Endotoxin induction of plasminogen activator and plasminogen activator inhibitor type 1 mRNA in rat tissues in vivo. J Biol Chem 1990; 265: 15560-15563
- 5 Darlington DN, Keil LC, Dallman MF. Potentiation of hormonal response to hemorrhage and fasting, but no hypoglycemia in conscious adrenalectomizedrats. Endocrinology 1989; 125: 1398-1406
- 6 Chomczynski P, Sacchii N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156-159
- 7 Bruzdzinski CJ, Riordan-Johnson M, Nordby EC, Sutter SM, Gelehrter TD. Isolation and characterization of the rat plasminogen activator inhibitor-1 gene. J Biol Chem 1990; 265: 2078-2085
- 8 Aird F, Clevenger CV, Prystowsky MB, Redei E. Corticotropin-releasing factor mRNA in rat thymus and spleen. Proc Natl Acad Sci USA 1993; 90: 7104-7108
- 9 Koji T, Brenner RM. Localization of estrogen receptor messenger ribonucleic acid in Rhesus monkey uterus by nonradioactive in situ hybridization with digoxigenin-labeled oligodeoxynucleotides Endocrinology . 1993; 132: 382-392
- 10 Schneiderman J, Sawdey M, Craig H, Thinnes T, Bordin TG, Loskutoff DJ. Type 1 plasminogen activator inhibitor gene expression following partial hepatectomy. Am J Pathol 1993; 143: 753-762
- 11 Nuovo GJ, Lindonnichi K, MacConnell P, Lane B. Intracellular localization of polymerase chain reaction (PCR)-amplified hepatitis C cDNA. JSurg Pathol 1993; 17: 683-690
- 12 Sawdey M, Podor TJ, Loskutoff DJ. Regulation of type 1 plasminogen activator inhibitor gene expression in cultured bovine aortic endothelial cells: induction by transforming growth factor-beta, lipopolysaccharide, and tumor necrosis factor-alpha. J Biol Chem 1989; 264: 10396-10401
- 13 Gann DS, Egdahl RH. Responses of adrenal corticosteroid secretion to hypotension and hypovolemia. J Clin Invest 1965; 44: 1-7
- 14 Ayala A, Wang P, Ba ZF, Perrin MM, Ertel W, Chaudry IH. Differential alterations in plasma IL-6 and TNF levels following trauma and hemorrhage. Am J Physiol 1991; 260: R167-171
- 15 Keeton M, Eguchi Y, Sawdey M, Ahn C, Loskutoff DJ. Cellular localization of type 1 plasminogen activator inhibitor messenger RNA and protein in murine renal tissue. Am J Pathol 1993; 142: 59-70
- 16 Komminoth P, Long AA. In-situ polymerase chain reaction. Virchows Archiv B Cell Pathol 1993; 64: 67-73
- 17 Whawell SA, Wang Y, Fleming KA, Thompson LM, Thompson JN. Localization ol’plasminogen activator inhibitor-1 production in inflamed appendix by insitu mRNA hybridization. J Pathol 1993; 169: 67-71
- 18 Idell S, Zwieh C, Kumar A, Koenig KB, Johnson AR. Pathways of fibrin turnover of human mesothelial cells in vitro. Am J Respir Cell Mol Biol 1992; 7: 414-426
- 19 Van llinshergh VW M, Kooistra T, Schaffer MA, van Bockel JH, van Muijen GNP. Charactcrization and fibrinolytic properties of human omental tissue mesothelial cell. Comparison with endothelial cells Blood 1990; 75: 1490-1497
- 20 Whawell SA, Scotl-Coombes DM, Vipond MN, Tebbutt SJ, Thompson JN. Tumor necrosis factor-mediated release of plasminogen activator inhibitor I by human peritoneal mesothelial cells. Brit J Surg 1994; 81: 214-216