Biochemical and functional characterization of glycosaminoglycans released from degranulating rat peritoneal mast cells: Insights into the physiological role of endogenous heparin.
The properties of commercially prepared heparin as an anticoagulant and antithrombotic agent in medicine are better understood than is the physiological role of heparin in its native form, where it is uniquely found in the secretory granules of mast cells. In the present study we have isolated and characterised the glycosaminoglycans (GAGs) released from degranulating rat peritoneal mast cells. Analysis of the GAGs by NMR spectroscopy showed the presence of both heparin and the galactosaminoglycan dermatan sulphate; heparinase digestion profiles and measurements of anticoagulant activity were consistent with this finding. The rat peritoneal mast cell GAGs significantly inhibited accumulation of leukocytes in the rat peritoneal cavity in response to IL-1β (p < 0.05, n = 6/group), and inhibited adhesion and diapedesis of leukocytes in the inflamed rat cremasteric microcirculation in response to LPS (p < 0.001, n = 4/group). FTIR spectra of human umbilical vein endothelial cells (HUVECs) were altered by treatment of the cells with heparin degrading enzymes, and restored by the addition of exogenous heparin. In conclusion, we have shown that rat peritoneal mast cells contain a mixture of GAGs that possess anticoagulant and anti-inflammatory properties.
UCL School of Pharmacy, Brunswick Square, London, WC1N 1AX, UK. Electronic address: rebecca.lever@ucl.ac.uk.
Rebecca
R
Lever
UCL School of Pharmacy, Brunswick Square, London, WC1N 1AX, UK; Sackler Institute of Pulmonary Pharmacology, Institute of Pharmaceutical Science, King's College London, London, SE1 9NH, UK.
Amir
A
Smailbegovic
Sackler Institute of Pulmonary Pharmacology, Institute of Pharmaceutical Science, King's College London, London, SE1 9NH, UK.
Yanira
Y
Riffo-Vasquez
Sackler Institute of Pulmonary Pharmacology, Institute of Pharmaceutical Science, King's College London, London, SE1 9NH, UK; National Institute for Biological Standards and Control, Potters Bar, Hertfordshire, EN6 3QG, UK.
Elaine
E
Gray
National Institute for Biological Standards and Control, Potters Bar, Hertfordshire, EN6 3QG, UK.
John
J
Hogwood
School of Chemistry, University of St. Andrews, St. Andrews, KY16 9ST, UK.
Stephen M
SM
Francis
School of Chemistry, University of St. Andrews, St. Andrews, KY16 9ST, UK.
Neville V
NV
Richardson
Sackler Institute of Pulmonary Pharmacology, Institute of Pharmaceutical Science, King's College London, London, SE1 9NH, UK. Electronic address: clive.page@kcl.ac.uk.
Clive P
CP
Page
Sackler Institute of Pulmonary Pharmacology, Institute of Pharmaceutical Science, King's College London, London, SE1 9NH, UK; National Institute for Biological Standards and Control, Potters Bar, Hertfordshire, EN6 3QG, UK.
Barbara
B
Mulloy
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000378:metabolism; D000925:Anticoagulants / Q000378:metabolism*; D003871:Dermatan Sulfate / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism*; D007962:Leukocytes / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008407:Mast Cells / Q000378:metabolism*; D008833:Microcirculation / Q000502:physiology; D010529:Peritoneal Cavity / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley
D000893:Anti-Inflammatory Agents; D000925:Anticoagulants; D006025:Glycosaminoglycans; D008070:Lipopolysaccharides; D003871:Dermatan Sulfate; D006493:Heparin
Dermatan sulphate; Glycosaminoglycan; Heparin; Mast cells