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Cultured Normal Human Hepatocytes do not Synthesize Lipoprotein-Associated Coagulation Inhibitor: Evidence that Endothelium is the Principal Site of Its Synthesis
Madhu S. Bajaj, Mohan N. Kuppuswamy, Hidehiko Saito, Silvia G. Spitzer and S. Paul Bajaj
Proceedings of the National Academy of Sciences of the United States of America
Vol. 87, No. 22 (Nov., 1990), pp. 8869-8873
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/2355648
Page Count: 5
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Human plasma contains a factor Xa-dependent inhibitor of tissue factor/factor VIIa complex termed lipoprotein-associated coagulation inhibitor (LACI). The present study examines the site(s) of LACI synthesis. In this study, cultured hepatocytes isolated from normal human liver were found to be essentially negative in LACI mRNA as revealed by Northern blot analysis using a full-length LACI cDNA as probe. The conditioned media from these cultures were also essentially negative for LACI activity. Similarly, poly(A)+ RNA obtained from normal human liver did not contain detectable LACI mRNA. In contrast, cultured human umbilical vein endothelial cells and human lung tissue (rich in endothelium) both contained abundant amounts of LACI mRNA. Moreover, erythrocyte lysates and culture media from normal monocytes, lymphocytes, or neutrophils did not contain measurable LACI activity; these cells were also negative for LACI mRNA. Platelets, however, contained LACI activity. The likely source of platelet LACI is the megakaryocyte cell since a megakaryocyte cell line (MEG-01) was found to contain LACI mRNA and to secrete small amounts of LACI activity. Additionally, human vascular smooth muscle cells and lung fibroblasts were also found to synthesize only small amounts of LACI. From these observations, we conclude that normal liver does not synthesize LACI and that endothelium is the principal source of plasma LACI. The undegraded LACI synthesized by endothelial cells had a molecular weight of ≈ 41,000.
Proceedings of the National Academy of Sciences of the United States of America © 1990 National Academy of Sciences