chondroitin-sulfates and Peritonitis

Fucosylated chondroitin sulfate

Topics: Animals; Anti-Inflammatory Agents; Cartilage; Chondroitin Sulfates; Circular Dichroism; Cucumaria; Disease Models, Animal; Female; Humans; Magnetic Resonance Spectroscopy; Molecular Structure; Peptones; Peritonitis; Rats; Rats, Wistar; Salmo salar; Structure-Activity Relationship; Treatment Outcome

Fucosylated chondroitin sulfate (fCS) extracted from the sea cucumber Holothuria forskali is composed of the following repeating trisaccharide unit: → 3)GalNAcβ4,6S(1 → 4) [FucαX(1 → 3)]GlcAβ(1 →, where X stands for different sulfation patterns of fucose (X = 3,4S (46%), 2,4S (39%), and 4S (15%)). As revealed by NMR and molecular dynamics simulations, the fCS repeating unit adopts a conformation similar to that of the Le(x) blood group determinant, bringing several sulfate groups into close proximity and creating large negative patches distributed along the helical skeleton of the CS backbone. This may explain the high affinity of fCS oligosaccharides for L- and P-selectins as determined by microarray binding of fCS oligosaccharides prepared by Cu(2+)-catalyzed Fenton-type and photochemical depolymerization. No binding to E-selectin was observed. fCS poly- and oligosaccharides display low cytotoxicity in vitro, inhibit human neutrophil elastase activity, and inhibit the migration of neutrophils through an endothelial cell layer in vitro. Although the polysaccharide showed some anti-coagulant activity, small oligosaccharide fCS fragments had much reduced anticoagulant properties, with activity mainly via heparin cofactor II. The fCS polysaccharides showed prekallikrein activation comparable with dextran sulfate, whereas the fCS oligosaccharides caused almost no effect. The H. forskali fCS oligosaccharides were also tested in a mouse peritoneal inflammation model, where they caused a reduction in neutrophil infiltration. Overall, the data presented support the action of fCS as an inhibitor of selectin interactions, which play vital roles in inflammation and metastasis progression. Future studies of fCS-selectin interaction using fCS fragments or their mimetics may open new avenues for therapeutic intervention.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Carbohydrate Conformation; Chondroitin Sulfates; Hydrogen Peroxide; Immune System Diseases; Iron; L-Selectin; Leukocyte Disorders; Leukocyte Elastase; Mice; Models, Molecular; Molecular Sequence Data; Neutrophil Infiltration; Neutrophils; Oxidation-Reduction; P-Selectin; Peritonitis; Proteinase Inhibitory Proteins, Secretory; Sea Cucumbers

Heparin is an excellent inhibitor of P- and L-selectin binding to the carbohydrate determinant, sialyl Lewis(x). As a consequence of its anti-selectin activity, heparin attenuates metastasis and inflammation. Here we show that fucosylated chondroitin sulfate (FucCS), a polysaccharide isolated from sea cucumber composed of a chondroitin sulfate backbone substituted at the 3-position of the beta-D-glucuronic acid residues with 2,4-disulfated alpha-L-fucopyranosyl branches, is a potent inhibitor of P- and L-selectin binding to immobilized sialyl Lewis(x) and LS180 carcinoma cell attachment to immobilized P- and L-selectins. Inhibition occurs in a concentration-dependent manner. Furthermore, FucCS was 4-8-fold more potent than heparin in the inhibition of the P- and L-selectin-sialyl Lewis(x) interactions. No inhibition of E-selectin was observed. FucCS also inhibited lung colonization by adenocarcinoma MC-38 cells in an experimental metastasis model in mice, as well as neutrophil recruitment in two models of inflammation (thioglycollate-induced peritonitis and lipopolysaccharide-induced lung inflammation). Inhibition occurred at a dose that produces no significant change in plasma activated partial thromboplastin time. Removal of the sulfated fucose branches on the FucCS abolished the inhibitory effect in vitro and in vivo. Overall, the results suggest that invertebrate FucCS may be a potential alternative to heparin for blocking metastasis and inflammatory reactions without the undesirable side effects of anticoagulant heparin.

Topics: Adenocarcinoma; Animals; Anticoagulants; Carbohydrate Conformation; Cell Adhesion; Chondroitin Sulfates; Disease Models, Animal; Dose-Response Relationship, Drug; Heparin; L-Selectin; Lipopolysaccharides; Lung Neoplasms; Mice; Neoplasm Metastasis; Neoplasms, Experimental; Neutrophil Infiltration; P-Selectin; Partial Thromboplastin Time; Peritonitis; Pneumonia; Sea Cucumbers; Thioglycolates