Liver sinusoidal endothelial cells are the principal site for elimination of unfractionated heparin from the circulation.
The mechanism of elimination of blood borne heparin was studied. To this end unfractionated heparin (UFH) was tagged with FITC, which served as both a visual marker and a site of labeling with (125)I-iodine. UFH labeled in this manner did not alter the anticoagulant activity or binding specificity of the glycosaminoglycan. Labeled heparin administered intravenously to rats (0.1 IU/kg) had a circulatory t(1/2) of 1.7 min, which was increased to 16 min upon coinjection with unlabeled UFH (100 IU/kg). At 15 min after injection, 71% of recovered radioactivity was found in liver. Liver cell separation revealed the following relative uptake capacity, expressed per cell: liver sinusoidal endothelial cell (LSEC)-parenchymal cell-Kupffer cell = 15:3.6:1. Fluorescence microscopy on liver sections showed accumulation of FITC-UFH only in cells lining the liver sinusoids. No fluorescence was detected in parenchymal cells or endothelial cells lining the central vein. Fluorescence microscopy of cultured LSECs following binding of FITC-UFH at 4 degrees C and chasing at 37 degrees C, showed accumulation of the probe in vesicles located at the periphery of the cells after 10 min, with transfer to large, evenly stained vesicles in the perinuclear region after 2 h. Immunogold electron microscopy of LSECs to probe the presence of FITC following injection of FITC-UFH along with BSA-gold to mark lysosomes demonstrated colocalization of the probe with the gold particles in the lysosomal compartment. Receptor-ligand competition experiments in primary cultures of LSECs indicated the presence of a specific heparin receptor, functionally distinct from the hyaluronan/scavenger receptor (Stabilin2). The results suggest a major role for LSECs in heparin elimination.
Center for Atherothrombotic Research in Tromsø, Department of Medicine, Institute of Clinical Medicine, University of Tromsø, N-9037 Tromsø, Norway. cristina@fagmed.uit.no
Cristina Ionica
CI
Oie
Randi
R
Olsen
Bård
B
Smedsrød
John-Bjarne
JB
Hansen
D000818:Animals; D000925:Anticoagulants / Q000097:blood / Q000378:metabolism*; D001693:Biological Transport, Active; D004705:Endocytosis / Q000502:physiology; D004847:Epithelial Cells / Q000378:metabolism* / Q000648:ultrastructure; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006493:Heparin / Q000097:blood / Q000378:metabolism*; D022781:Hepatocytes / Q000378:metabolism* / Q000648:ultrastructure; D007150:Immunohistochemistry; D008297:Male; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D051381:Rats; D017207:Rats, Sprague-Dawley; D014018:Tissue Distribution
D000925:Anticoagulants; D005456:Fluorescent Dyes; D006493:Heparin; D016650:Fluorescein-5-isothiocyanate