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Thrombosis Research 2008Dermatan sulfate, a sulfated glycosaminoglycan, acts as an anticoagulant by accelerating the inhibition of thrombin by heparin cofactor II.
INTRODUCTION
Dermatan sulfate, a sulfated glycosaminoglycan, acts as an anticoagulant by accelerating the inhibition of thrombin by heparin cofactor II.
MATERIALS AND METHODS
A low molecular mass dermatan sulfate was obtained by peroxy-radical depolymerization from a parent dermatan sulfate. Chemical characterization of this low molecular mass dermatan sulfate shows a material of approximately 5 kDa that conserves sulfated sequences (2-O-sulfation of the iduronic acid units and/or 4 or 6 positions of galactosamina N acetyl) essential for dermatan sulfate-heparin cofactor II interaction with more sulphated proportion (27.7+/-1.9 microg% vs 11.5+/-0.8 microg%, P<0.05 n=6, low molecular mass dermatan sulfate vs dermatan sulfate).
RESULTS
After a single intravenous administration of low molecular dermatan sulfate in rats, fibrinolytic activity increased simultaneously with thrombin clotting time prolongation. Low molecular dermatan sulfate showed an inhibitory effect on classical complement activation pathway reaching a maximum during the first hour. Furthermore, low molecular dermatan sulfate was as effective as dermatan sulfate to prevent thrombus formation and to diminish thrombus weight in a rat venous thrombosis model.
CONCLUSIONS
The results indicate that peroxy-radical depolymerization of dermatan sulfate produced a low molecular dermatan sulfate with profibrinolytic, thrombolytic, antithrombotic and anticomplement properties. We conclude that low molecular dermatan sulfate may be an effective adjunct in the management of thrombotic events.
Topics: Animals; Anticoagulants; Complement System Proteins; Dermatan Sulfate; Fibrinolytic Agents; Hemolysis; Male; Molecular Weight; Rats; Rats, Wistar; Thrombin
PubMed: 17936880
DOI: 10.1016/j.thromres.2007.09.001 -
Methods in Molecular Biology (Clifton,... 2010Chondroitin/dermatan sulfate (CS/DS) glycosaminoglycans (GAGs) are present in high levels in connective tissue where they play roles as structural molecules and in...
Chondroitin/dermatan sulfate (CS/DS) glycosaminoglycans (GAGs) are present in high levels in connective tissue where they play roles as structural molecules and in protein-binding interactions. Recent developments in the techniques for analysis of CS/DS using capillary electrophoresis (CE) have enabled progress in the understanding of changes in CS/DS structure that accompany connective tissue diseases including osteoarthritis. Key to these developments is the ability to extract CS/DS GAGs from small quantities of connective tissue. This chapter describes a method for connective tissue GAG extraction, derivatization, and workup for subsequent capillary electrophoretic and/or mass spectrometric analysis.
Topics: Adult; Child; Chondroitin; Chondroitin Sulfates; Chromatography, Ion Exchange; Connective Tissue; Dermatan Sulfate; Electrophoresis, Capillary; Humans; Mass Spectrometry; Molecular Biology; Oligosaccharides; Osteoarthritis
PubMed: 19882131
DOI: 10.1007/978-1-60761-454-8_15 -
Glycoconjugate Journal Feb 2021Oviductus ranae (O.ran.) has been widely used as a tonic and a traditional animal-based Chinese medicine. O.ran. extracts have been reported to have numerous biological...
Oviductus ranae (O.ran.) has been widely used as a tonic and a traditional animal-based Chinese medicine. O.ran. extracts have been reported to have numerous biological activities, including activities that are often associated with mammalian glycosaminoglycans such as anti-inflammatory, antiosteoperotic, and anti-asthmatic. Glycosaminoglycans are complex linear polysaccharides ubiquitous in mammals that possess a wide range of biological activities. However, their presence and possible structural characteristics within O.ran. were previously unknown. In this study, glycosaminoglycans were isolated from O.ran. and their disaccharide compositions were analyzed by liquid chromatography-ion trap/time-of-flight mass spectrometry (LC-MS-ITTOF). Heparan sulfate (HS)/heparin (HP), chondroitin sulfate (CS)/dermatan sulfate (DS) and hyaluronic acid (HA) were detected in O.ran. with varied disaccharide compositions. HS species contain highly acetylated disaccharides, and have various structures in their constituent chains. CS/DS chains also possess a heterogeneous structure with different sulfation patterns and densities. This novel structural information could help clarify the possible involvement of these polysaccharides in the biological activities of O.ran..
Topics: Chondroitin Sulfates; Chromatography, Liquid; Dermatan Sulfate; Disaccharides; Glycosaminoglycans; Heparin; Heparitin Sulfate; Mass Spectrometry; Materia Medica; Sensitivity and Specificity
PubMed: 33411075
DOI: 10.1007/s10719-020-09962-8 -
Carbohydrate Research Sep 1993Heparin from beef intestinal mucosa, dermatan sulfate from beef intestinal mucosa, and chondroitin sulfate from bovine trachea were extracted and purified, and their...
Heparin from beef intestinal mucosa, dermatan sulfate from beef intestinal mucosa, and chondroitin sulfate from bovine trachea were extracted and purified, and their structures and physico-chemical properties were evaluated by different techniques (disaccharide patterns by specific enzymatic cleavage, relative molecular mass by high-performance size-exclusion chromatography, sulfate-to-carboxyl ratio by potentiometric determination). Heparin was fractionated into "slow moving" and "fast moving" fractions by selective precipitation as the barium salt at different temperatures. The "fast moving" and "slow moving" components of heparin, dermatan sulfate, and chondroitin sulfate were utilized to run calibration curves in agarose-gel electrophoresis. Mixtures containing different amounts of these glycosaminoglycans were made and separated by agarose-gel electrophoresis, and these were analyzed quantitatively. For analysis of relative amounts, the area of each individual component of mixtures, obtained by photodensitometric readings, was divided by the sum of the areas of all glycosaminoglycans and expressed as a percentage. For analysis of absolute amounts, the area under the curve for each component of mixtures was fitted to specific calibration curves, and the amount of each glycosaminoglycan was calculated in micrograms. The quantitative procedure performed by analysing absolute amounts was used to obtain an accurate quantitative evaluation of each component in mixtures of glycosaminoglycans utilized for pharmaceutical purposes. A sensitive method was developed for the evaluation of very small amounts (0.2% w/w) of possible glycosaminoglycans as contaminants in preparations of a single species of glycosaminoglycan. This technique requires specific enzymatic degradation by bacterial lyases, separation in agarose-gel electrophoresis, and quantitative analysis by photodensitometric analysis and specific calibration curves.
Topics: Animals; Carbohydrate Sequence; Cattle; Chondroitin Sulfates; Chromatography, High Pressure Liquid; Dermatan Sulfate; Disaccharides; Electrophoresis, Agar Gel; Heparin; Intestinal Mucosa; Molecular Sequence Data; Molecular Weight; Mucous Membrane; Optical Rotation; Trachea
PubMed: 8221721
DOI: 10.1016/0008-6215(93)84259-9 -
Glycobiology Jul 2011Chondroitin sulfate (CS)/dermatan sulfate (DS) is a group of sulfated polymers, which play an essential role in various biological phenomena. In the kidney, they are...
Chondroitin sulfate (CS)/dermatan sulfate (DS) is a group of sulfated polymers, which play an essential role in various biological phenomena. In the kidney, they are present in small but significant amounts. Studies on their structure-function relationship in the kidney and their changes during diabetic conditions have not been rigorously looked into, which is the focus of this paper. The CS/DS content decreased significantly (14%) during diabetic conditions. This was accompanied by a decrease in the CS/heparan sulfate ratio. Disaccharide composition analysis revealed fine structural changes especially with respect to the E unit [glucuronic acid β1-3 N-acetyl d-galactosamine (4,6-O-sulfate)] and the degree of sulfation. The mRNA expression levels of major enzymes involved in the synthesis of the "E"-disaccharide unit showed a decrease during diabetes. The changes in CS/DS had implications on ligand-binding properties when tested in vitro for binding to major extracellular matrix (ECM) components such as type IV collagen, laminin and fibronectin. Thus, this study provides insights into the structure-function relationship of CS/DS in the kidney during diabetes and alterations of which could aggravate conditions such as diabetic nephropathy by virtue of them being a part of ECM components.
Topics: Animals; Blotting, Western; Chondroitin Sulfates; Dermatan Sulfate; Diabetes Complications; Diabetes Mellitus, Experimental; Extracellular Matrix; Kidney; Male; RNA, Messenger; Rats; Rats, Wistar; Reverse Transcriptase Polymerase Chain Reaction; Sulfotransferases
PubMed: 21406563
DOI: 10.1093/glycob/cwr029 -
Analytical Biochemistry Dec 2014We developed a method using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) with a selected reaction monitoring (SRM) mode for...
Quantitative analysis of glycosaminoglycans, chondroitin/dermatan sulfate, hyaluronic acid, heparan sulfate, and keratan sulfate by liquid chromatography-electrospray ionization-tandem mass spectrometry.
We developed a method using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) with a selected reaction monitoring (SRM) mode for simultaneous quantitative analysis of glycosaminoglycans (GAGs). Using one-shot analysis with our MS/MS method, we demonstrated the simultaneous quantification of a total of 23 variously sulfated disaccharides of four GAG classes (8 chondroitin/dermatan sulfates, 1 hyaluronic acid, 12 heparan sulfates, and 2 keratan sulfates) with a sensitivity of less than 0.5 pmol within 20 min. We showed the differences in the composition of GAG classes and the sulfation patterns between porcine articular cartilage and yellow ligament. In addition to the internal disaccharides described above, some saccharides derived from the nonreducing terminal were detected simultaneously. The simultaneous quantification of both internal and nonreducing terminal saccharides could be useful to estimate the chain length of GAGs. This method would help to establish comprehensive "GAGomic" analysis of biological tissues.
Topics: Animals; Cartilage, Articular; Chondroitin Sulfates; Chromatography, Liquid; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate; Hyaluronic Acid; Keratan Sulfate; Spectrometry, Mass, Electrospray Ionization; Swine; Tandem Mass Spectrometry
PubMed: 25197028
DOI: 10.1016/j.ab.2014.08.005 -
Scientific Reports Feb 2020Glycosaminoglycans (GAGs) are polysaccharides produced by most mammalian cells and involved in a variety of biological processes. However, due to the size and complexity...
Glycosaminoglycans (GAGs) are polysaccharides produced by most mammalian cells and involved in a variety of biological processes. However, due to the size and complexity of GAGs, detailed knowledge about the structure and expression of GAGs by cells, the glycosaminoglycome, is lacking. Here we report a straightforward and versatile approach for structural domain mapping of complex mixtures of GAGs, GAGDoMa. The approach is based on orthogonal enzymatic depolymerization of the GAGs to generate internal, terminating, and initiating domains, and nanoflow reversed-phase ion-pairing chromatography with negative mode higher-energy collision dissociation (HCD) tandem mass spectrometry (MS/MS) for structural characterization of the individual domains. GAGDoMa provides a detailed structural insight into the glycosaminoglycome, and offers an important tool for deciphering the complexity of GAGs in cellular physiology and pathology.
Topics: Carbohydrate Sequence; Cell Line; Chondroitin Sulfates; Chromatography, High Pressure Liquid; Chromatography, Reverse-Phase; Dermatan Sulfate; Glycosaminoglycans; Humans; Tandem Mass Spectrometry
PubMed: 32103093
DOI: 10.1038/s41598-020-60526-0 -
Journal of Pharmaceutical and... Jul 2018Sulodexide is a heparinoid which combines the properties of its components heparin and dermatan sulfate and is used not only for the prophylaxis and treatment of...
Sulodexide is a heparinoid which combines the properties of its components heparin and dermatan sulfate and is used not only for the prophylaxis and treatment of thromboembolic diseases but also for the treatment of diabetic nephropathy. Despite many clinical studies have been conducted to investigate its activity and safety, no data are available on the fine chemical characterization of its components. In this work, the in-depth investigation on the structural features of both the whole mixture and the isolated components was accomplished, involving the analysis of molecular weight distribution and of their mono, di and oligosaccharide composition by HP-SEC/TDA, 2D-NMR and HPLC-MS techniques. Moreover, also the separation of fractions endowed of graded affinity to antithrombin was achieved followed again by detailed structural analysis. The combination of different techniques permits to profile in depth the structural features of such a drug and offers a useful tool for possible analysis of batch production.
Topics: Chemical Fractionation; Chromatography, High Pressure Liquid; Dermatan Sulfate; Glycosaminoglycans; Heparin; Magnetic Resonance Spectroscopy; Mass Spectrometry; Molecular Structure; Molecular Weight; Oligosaccharides
PubMed: 29689470
DOI: 10.1016/j.jpba.2018.04.012 -
Blood Oct 1989Dermatan sulfate (DS), a catalyst of the thrombin-heparin cofactor II interaction, has antithrombotic activity and is devoid of significant hemorrhagic risk in several...
Dermatan sulfate (DS), a catalyst of the thrombin-heparin cofactor II interaction, has antithrombotic activity and is devoid of significant hemorrhagic risk in several animal models. We investigated the pharmacodynamic and pharmacokinetic properties of DS in humans. DS was injected in single bolus intravenous injections of four increasing doses (0.5, 1, 1.5, 2 mg/kg) to six healthy volunteers. The resulting anticoagulant activities were assessed by the activated partial thromboplastin time (APTT) and the thrombin clotting time (TCT). There were dose-dependent prolongations of the APTT and TCT, and the anticoagulant activities disappeared in less than three hours. The pharmacokinetic parameters were calculated from the plasma concentrations of DS measured with a new chromogenic assay. The volume of distribution was approximately 1.8 times greater than the theoretical plasma volume and was independent of dose. In contrast, the clearance decreased with dose and the terminal half-life ranged from 0.45 +/- 0.08 hours at 0.5 mg/kg to 0.72 +/- 0.11 hours (mean +/- SD) at 2 mg/kg. The bioavailabilities of subcutaneous (SC) and intramuscular (IM) administration relative to those of intravenous administration were determined in 12 other volunteers. The respective bioavailabilities were 24.7% +/- 12.9% and 12.4% +/- 9.2% for SC and IM administration. There was no detectable change in the APTT and the TCT when the volunteers were injected with 1.5 mg/kg SC or IM. In addition, the pharmacokinetic parameters derived from plasma concentrations of DS showed considerable interindividual variations by the two later routes of administration. Peak concentrations were noted 2.7 +/- 1.3 hours after SC injection and 4.3 +/- 4.9 hours after IM injection. The average peak concentrations were 0.7 +/- 0.3 and 0.4 +/- 0.2 mg/L after SC and IM injections, respectively. The half-lives of DS were 7.9 +/- 6.5 hours (SC) and 6.3 +/- 7.4 hours (IM). No adverse reaction to DS was recorded during this study.
Topics: Adult; Blood Coagulation; Chondroitin; Dermatan Sulfate; Dose-Response Relationship, Drug; Female; Humans; Male; Metabolic Clearance Rate; Partial Thromboplastin Time; Thrombin Time
PubMed: 2790187
DOI: No ID Found -
Biological & Pharmaceutical Bulletin 2010We examined the effects of chondroitinases on the release of dermatan sulfate (DS)-induced arginine amidase (AA) from rabbit ear artery. DS-induced AA release was...
We examined the effects of chondroitinases on the release of dermatan sulfate (DS)-induced arginine amidase (AA) from rabbit ear artery. DS-induced AA release was significantly decreased by treatment with chondroitinase ABC (ABCase) in the rabbit ear artery. On the other hand, Chondroitinase ACII (ACIIase) enhanced spontaneous and DS-induced AA release. Heat-inactivated ABCase and ACIIase did not affect spontaneous and DS-induced AA release. Furthermore, ABCase, but not ACIIase and heat-inactivated chondroitinases, degraded DS. These results indicate that the facilitatory effect of DS-induced AA release from the rabbit ear artery is affected by the molecular size of DS.
Topics: Animals; Anticoagulants; Arteries; Chondroitin ABC Lyase; Chondroitin Lyases; Dermatan Sulfate; Ear; Rabbits; Serine Endopeptidases; Structure-Activity Relationship
PubMed: 20045955
DOI: 10.1248/bpb.33.150