chondroitin-sulfates and Arteriosclerosis

Thromboembolic vascular diseases remain the main cause of death in Western industrialized societies. Anticoagulants retard the formation, growth, and embolization of thrombi and are effective agents in the prevention and treatment of thromboembolic disease. Anticoagulants in venous thromboembolism have been investigated extensively with rigorous randomized, controlled trials, while the roles for anticoagulants in arterial thromboembolism generally have evolved through natural history studies and empirical practice. Thus, many current guidelines for anticoagulant use in arterial disease are based on successful established routines and rational therapy. To effectively balance the efficacy and risks of anticoagulation, the vascular surgeon needs a thorough understanding of anticoagulant drugs, their mechanisms of action, and their proven and unproven indications. Since the first use of heparin in arterial surgery, a variety of new and different anticoagulants have become available, including low-molecular-weight heparins, heparin-like drugs, hirudins, and thrombin inhibitors. Despite their diverse actions, they all inhibit some portion of the plasma coagulation cascade, thus distinguishing them from platelet inhibitors or fibrinolytics. Every interference with the coagulation cascade carries a risk of minor, major, or fatal hemorrhage. To date, no drug or therapeutic strategy has succeeded fully in dissociating its antithrombotic effects from its risks of bleeding.

Topics: Anticoagulants; Arteriosclerosis; Blood Coagulation; Chondroitin Sulfates; Coumarins; Dermatan Sulfate; Drug Combinations; Heparin; Heparin, Low-Molecular-Weight; Heparitin Sulfate; Humans; Platelet Activation; Thrombin; Thromboembolism

Because the role of elemental sulfur in human nutrition has not been studied extensively, it is the purpose of this article to emphasize the importance of this element in humans and discuss the therapeutic applications of sulfur compounds in medicine. Sulfur is the sixth most abundant macromineral in breast milk and the third most abundant mineral based on percentage of total body weight. The sulfur-containing amino acids (SAAs) are methionine, cysteine, cystine, homocysteine, homocystine, and taurine. Dietary SAA analysis and protein supplementation may be indicated for vegan athletes, children, or patients with HIV, because of an increased risk for SAA deficiency in these groups. Methylsulfonylmethane (MSM), a volatile component in the sulfur cycle, is another source of sulfur found in the human diet. Increases in serum sulfate may explain some of the therapeutic effects of MSM, DMSO, and glucosamine sulfate. Organic sulfur, as SAAs, can be used to increase synthesis of S-adenosylmethionine (SAMe), glutathione (GSH), taurine, and N-acetylcysteine (NAC). MSM may be effective for the treatment of allergy, pain syndromes, athletic injuries, and bladder disorders. Other sulfur compounds such as SAMe, dimethylsulfoxide (DMSO), taurine, glucosamine or chondroitin sulfate, and reduced glutathione may also have clinical applications in the treatment of a number of conditions such as depression, fibromyalgia, arthritis, interstitial cystitis, athletic injuries, congestive heart failure, diabetes, cancer, and AIDS. Dosages, mechanisms of action, and rationales for use are discussed. The low toxicological profiles of these sulfur compounds, combined with promising therapeutic effects, warrant continued human clinical trails.

Topics: Amino Acids, Essential; Antioxidants; Arteriosclerosis; Chondroitin Sulfates; Diet; Dimethyl Sulfoxide; Glucosamine; Glutathione Transferase; HIV Infections; Humans; Muscle Fatigue; Sulfur; Thioctic Acid

Topics: Amino Acid Sequence; Animals; Arteries; Arteriosclerosis; Carbohydrate Sequence; Chondroitin Sulfates; Columbidae; Dermatan Sulfate; Humans; Hyaluronic Acid; In Vitro Techniques; Macromolecular Substances; Models, Molecular; Molecular Weight; Proteoglycans

The etiology of ischemic stroke affects prognosis, outcome, and management. Trials of therapies for patients with acute stroke should include measurements of responses as influenced by subtype of ischemic stroke. A system for categorization of subtypes of ischemic stroke mainly based on etiology has been developed for the Trial of Org 10172 in Acute Stroke Treatment (TOAST).. A classification of subtypes was prepared using clinical features and the results of ancillary diagnostic studies. "Possible" and "probable" diagnoses can be made based on the physician's certainty of diagnosis. The usefulness and interrater agreement of the classification were tested by two neurologists who had not participated in the writing of the criteria. The neurologists independently used the TOAST classification system in their bedside evaluation of 20 patients, first based only on clinical features and then after reviewing the results of diagnostic tests.. The TOAST classification denotes five subtypes of ischemic stroke: 1) large-artery atherosclerosis, 2) cardioembolism, 3) small-vessel occlusion, 4) stroke of other determined etiology, and 5) stroke of undetermined etiology. Using this rating system, interphysician agreement was very high. The two physicians disagreed in only one patient. They were both able to reach a specific etiologic diagnosis in 11 patients, whereas the cause of stroke was not determined in nine.. The TOAST stroke subtype classification system is easy to use and has good interobserver agreement. This system should allow investigators to report responses to treatment among important subgroups of patients with ischemic stroke. Clinical trials testing treatments for acute ischemic stroke should include similar methods to diagnose subtypes of stroke.

Topics: Anticoagulants; Arteriosclerosis; Brain Ischemia; Cerebral Infarction; Chondroitin Sulfates; Dermatan Sulfate; Diagnosis, Differential; Embolism; Glycosaminoglycans; Heparinoids; Heparitin Sulfate; Humans

Effects of isomers of chondroitin sulfate on atherosclerosis were clinically compared, based on sulfate linkage and the amount of sulfate, by using chondroitin 4-sulfate, chondroitin 6-sulfate and chondroitin polysulfate. Fourty eight age-matched atherosclerotic subjects were selected from a home for the elderly in order to the treatment with the agents. The isomers of chondroitin sulfate were given a daily dose of 4.5 g perorally. During the experimental period for 64 months, mortality, serum cholesterol, thrombus-formation time and thrombus weight were examined. The result obtained was as follows: mortality in the groups treated with the isomers of chondroitin sulfate was less than the age-matched untreated control group. Serum cholesterol value in the group treated by the isomers of chondroitin sulfate, chondroitin polysulfate group in particular, fell lower than the pre-treatment value. Thrombus formation time prolonged 150% in the group treated with chondroitin polysulfate over the untreated control group and the resultant thrombus weight was reduced in the treated group. Thus, these data indicated that the isomers of chondroitin sulfate are clinically effective on the treatment of atherosclerosis in the order of chondroitin polysulfate, chondroitin 4-sulfate and/or chondroitin 6-sulfate.

Topics: Aged; Arteriosclerosis; Blood Coagulation; Cholesterol; Chondroitin; Chondroitin Sulfates; Clinical Trials as Topic; Humans; Isomerism; Thrombosis

Changes in chondroitin sulfate (CS) proteoglycan (PG) during atherosclerosis are associated with chronic inflammatory changes and increased incidence of thrombosis. To explore how glycosaminoglycan changes could influence the thrombogenicity of atherosclerotic lesions, water-transfer reactions were examined during activation of antithrombin by CS.. Advanced type IV atherosclerotic lesions prone to thrombosis contained CSPG (versican) with undersulfated CS relative to CS of the adjacent healthy aorta. Approximately 11% of the CS disaccharide in versican from healthy arteries was oversulfated, but this proportion decreased markedly to 3% in atherosclerotic lesions. Oversulfated CS functionally bound antithrombin with a dissociation constant of 3.3+/-1.9 micromol/L. Measured by osmotic stress (OS) techniques with an approximately 26-A probe, the reaction was linked to transfer of approximately 2500 mol water per mole of coagulation factor Xa inhibited. Under OS, the anticoagulant efficiency of CS was 1.3 (micromol/L)(-1) x s(-1), approximately 5- and 15-fold higher than heparan sulfate efficiency measured under OS and standard conditions, respectively.. Decreased sulfation of high molecular weight CSPG in the advancing atherosclerotic lesions may predispose the lesions to thrombosis by disrupting osmotic regulation, limiting avidity for antithrombin and decreasing activation efficiency.

Topics: Anticoagulants; Aorta; Arteriosclerosis; Chondroitin Sulfate Proteoglycans; Chondroitin Sulfates; Factor Xa; Glycosaminoglycans; Humans; Lectins, C-Type; Osmotic Pressure; Proteoglycans; Versicans; Water

Topics: Antithrombins; Arteriosclerosis; Blood Coagulation; Chondroitin Sulfate Proteoglycans; Chondroitin Sulfates; Extracellular Matrix; Glycosaminoglycans; Homeostasis; Lectins, C-Type; Osmotic Pressure; Versicans; Water

Glycosaminoglycans participate in several biological functions in the arterial wall through their specific structures. They undergo specific compositional and structural modifications during the development of vascular diseases. The present study was performed to determine the variations in the concentration and the structural characteristics of galactosaminoglycans--chondroitin sulfate (CS) and dermatan sulfate (DS)--during the progression of atherosclerosis and aneurysmal dilatation of the human abdominal aorta. The concentration of CS was increased 24% (p < or = 0.05) in atherosclerotic type II aortas, but it was significantly decreased (29%, p < or = 0.05) in atherosclerotic type V aortas and aneurysmal aortas (65%, p < or = 0.01). In contrast, the concentration of DS was almost constant in all stages of arterial disease examined. Significant structural alterations were detected in the disaccharide composition of galactosaminoglycans. The ratio of 6-sulfated to 4-sulfated disaccharides was increased in atherosclerotic type II aortas (4.0 instead of 3.1 in normal aortas) due to the marked increase of CS in this tissue. This ratio was significantly decreased in atherosclerotic type V and aneurysmal aortas (2.1 and 1.6, respectively) due to the significant reduction of CS in the respective tissues. In addition, significant decrease of the oversulfated disaccharides, which are mainly located in DS chains, was recorded in atherosclerotic and aneurysmal aortas. Particularly, deltadi-di(2,6)S were decreased 32% (p < or = 0.01) and 86% (p < or = 0.01) in atherosclerotic type II and V aortas and 88% (p < or = 0.01) in aneurysm. Deltadi-di(2,4)S were increased in atherosclerotic type II aortas (21%, p < or = 0.01), but significantly decreased in atherosclerotic type V (33%, p < or = 0.01) and aneurysmal aortas (56%, p < or = 0.01). The amounts of deltadi-di(4,6)S were not markedly affected in the diseased tissues. These results suggest that the concentration of galactosaminoglycans is differentially affected during the progression of atherosclerosis. Furthermore, the development of vascular disease is associated with specific structural modifications, especially with the significant reduction of particular types of oversulfated disaccharides, which may play essential biological roles in the arterial wall.

Topics: Adult; Aorta, Abdominal; Aortic Aneurysm, Abdominal; Arteriosclerosis; Chemical Fractionation; Chondroitin ABC Lyase; Chondroitin Lyases; Chondroitin Sulfates; Dermatan Sulfate; Disaccharides; Electrophoresis, Capillary; Humans; Male; Middle Aged; Uronic Acids

Macrophages are intimately involved in the pathogenesis of atherosclerotic diseases. A key feature of this process is their uptake of various lipoproteins and subsequent transformation to foam cells. Since lipoprotein lipase (LPL) is believed to play a role in foam cell formation, we investigated if endogenously produced proteoglycans (PGs) affect the release of this enzyme from macrophages. The human leukaemic cell line THP-1 which differentiates into macrophages by treatment with phorbol ester (phorbol 12-myristate 13-acetate) served as a model. The differentiation of THP-1 macrophages promoted the release of PGs into the cell medium which caused the detachment of LPL activity from the cell surface, and prevented LPL re-uptake and inactivation. These PGs were mainly composed of chondroitin sulfate type and exerted a heparin-like effect on LPL release. LPL is known to increase the cell association of lipoproteins by the well known bridging function. Exogenous bovine LPL at a concentration of 1 microg/ml enhanced low density lipoprotein (LDL)-binding 10-fold. Endogenously produced PGs reduced LPL-mediated binding of LDL. It is proposed that the differentiation-dependent increase in the release of PGs interferes with binding of LPL and reduces lipoprotein-binding to macrophages.

Topics: Arteriosclerosis; Cell Line; Chondroitin Sulfates; Culture Media, Conditioned; Dermatan Sulfate; Glycosaminoglycans; Heparin; Humans; Lipoprotein Lipase; Lipoproteins; Lipoproteins, LDL; Macrophages; Proteoglycans

Aggregated low-density lipoprotein (LDL) was shown to be present in the atherosclerotic lesion, but the mechanism responsible for its formation in vivo is not known yet. To find out whether LDL aggregation occurs in the arterial wall during atherogenesis, LDLs were extracted from the aortas of apolipoprotein E-deficient (E(0)) mice during their aging (and the development of atherosclerosis), and were analyzed for their aggregation states, in comparison to LDLs isolated from aortas of control mice. LDL isolated from aortas of E(0) mice was already aggregated at 1 month of age and its aggregation state substantially increased with age, with 3-fold elevation at 6 months of age compared to younger, 1-month-old, mice. Only minimal aggregation could be detected in LDL derived from control mice. Electron microscopy examination revealed that LDL particles from aortas of the E(0) mice were heterogeneous in their size, ranging between 20 and 300 nm. The mouse aortic LDL contained proteoglycans (PGs) and their content increased with the age of the mice, with about 2-fold higher levels than those found in LDLs derived from aortas of control mice. Macrophage-released PGs were previously demonstrated to enhance LDL aggregation in vitro. However, their involvement in LDL aggregation in vivo has not been studied yet. Thus, we next studied the effect of arterial macrophage-released PGs on the susceptibility of plasma LDL to aggregation by Bacillus cereus sphingomyelinase (SMase). Foam cell macrophages were isolated from aortas of the atherosclerotic E(0) mice at 6 months of age and were found to be loaded with cholesterol and to contain oxidized lipids. To analyze the effect of macrophage-released PGs on LDL aggregation, PGs were prelabeled by cell incubation with [35S]sulfate, followed by incubation of macrophage-released PGs with E(0) mouse plasma LDL (200 microg protein/ml) for 1 h at 37 degrees C. [35S]Sulfated PGs were found to be LDL-associated and the susceptibility of PG-associated LDL to aggregation by SMase was increased by up to 45% in comparison to control LDL. Similar results demonstrating the involvement of PGs in LDL aggregation were obtained upon incubation of LDL with increasing concentrations of PGs that were isolated from the entire aorta of E(o) mice (rather than the isolated macrophages). The stimulatory effect of macrophage-released PGs on LDL aggregation was markedly reduced when the PGs were pretreated with the glycosaminoglycan-hydrolyzing enzymes

Topics: Aging; Animals; Aorta; Apolipoproteins E; Arteriosclerosis; Chondroitin Sulfates; Heparitin Sulfate; Lipoproteins, LDL; Macrophages; Mice; Microscopy, Electron; Particle Size; Proteoglycans

Lp(a) interference with fibrinolysis could contribute to atherothrombosis. Additionally, accumulation of Lp(a) and LDLs, could lead to cholesterol deposition and foam cell formation in atherogenesis. The interactions between Lp(a) and LDL could cause their entrapment in the extracellular matrix of lesions. We found that association of Lp(a) with matrix secreted by cultured human arterial smooth muscle cells increased 2 to 3 times the subsequent specific binding of radioactive LDL. Chondroitin sulfate proteoglycans seem responsible for formation of the specific matrix-Lp(a) and matrix-LDL aggregates. The proteoglycans appeared also to participate in a cooperative increase of radioactive LDL binding to matrix pretreated with Lp(a). In the matrix preincubated with LDL, approximately 50% of the additional lipoprotein was bound by ionic interactions. In the matrix preincubated with Lp(a), 20% of the additional LDL was held by ionic bonds, and the rest was held by strong nonionic associations. Binding analysis in physiological solutions confirmed that chondroitin sulfate-rich proteoglycans from the smooth muscle cell matrix have a high affinity for Lp(a) and LDL. The results provide an explanation to the observed localization of Lp(a) and LDL in the extracellular matrix of arterial lesions and suggest a mechanism for their cooperative accumulation there.

Topics: Arteriosclerosis; Cells, Cultured; Chondroitin Sulfates; Dermatan Sulfate; Extracellular Matrix; Fibrinolysis; Humans; Lipoprotein(a); Lipoproteins, LDL; Muscle, Smooth, Vascular; Protein Binding; Proteoglycans

Atherosclerotic lesions contain an extracellular sphingomyelinase (SMase) activity that hydrolyzes the sphingomyelin of subendothelial low density lipoprotein (LDL). This SMase activity may promote atherosclerosis by enhancing subendothelial LDL retention and aggregation, foam cell formation, and possibly other atherogenic processes. The results of recent cell-culture studies have led to the hypothesis that a specific molecule called secretory SMase (S-SMase) is responsible for the SMase activity known to be in lesions, although its presence in atheromata had not been examined directly. Herein we provide immunohistochemical and biochemical support for this hypothesis. First, 2 different antibodies against S-SMase detected extracellular immunoreactive protein in the intima of mouse, rabbit, and human atherosclerotic lesions. Much of this material in lesions appeared in association with the subendothelial matrix. Second, binding studies in vitro demonstrated that (125)I-S-SMase adheres to the extracellular matrix of cultured aortic smooth muscle and endothelial cells, specifically to the laminin and collagen components. Third, in its bound state, S-SMase retains substantial enzymatic activity against lipoprotein substrates. Overall, these data support the hypothesis that S-SMase is an extracellular arterial wall SMase that contributes to the hydrolysis of the sphingomyelin of subendothelial LDL. S-SMase may therefore be an important participant in atherogenesis through local enzymatic effects that stimulate subendothelial retention and aggregation of atherogenic lipoproteins.

Topics: Animals; Aorta; Arteriosclerosis; Cattle; Cells, Cultured; Chondroitin Sulfates; Collagen; Dermatan Sulfate; Endothelium, Vascular; Extracellular Matrix Proteins; Female; Fibronectins; Heparin; Humans; Iodine Radioisotopes; Laminin; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Muscle, Smooth, Vascular; Protein Binding; Rabbits; Serum Albumin, Bovine; Sphingomyelin Phosphodiesterase

Heparan sulfate is thought to regulate the biological activities of several proteins implicated in the pathogenesis of atherosclerosis. While the interactions of heparan sulfate with lipoprotein lipase and various growth factors have been actively studied, little is known of the cellular regulation of heparan sulfate biosynthesis in response to lipid accumulation. We have investigated heparan sulfate biosynthesis during conversion of murine J774 macrophages into lipid-laden foam cells. Such conversion is shown to accelerate the rate of glycosaminoglycan synthesis and the transport of newly synthesized proteoglycans into the medium. Moreover, the structure of heparan sulfate is specifically altered due to an approximately 30% increase in the 6-O-sulfation of glucosamine residues within the N-sulfated heparan sulfate domains, whereas the sulfation of chondroitin sulfate remains unaffected. These results suggest a selective effect of foam cell conversion on the biosynthesis of heparan sulfate.

Topics: Animals; Arteriosclerosis; Cell Line; Chondroitin Sulfates; Disaccharides; Foam Cells; Glycosaminoglycans; Heparitin Sulfate; Humans; Lipoproteins, LDL; Macrophages; Mice; Nitrous Acid; Oligosaccharides

The walls of human abdominal aortas and atherosclerosis-induced aneurysms contain similar amounts of collagen. The quantitative ratio between collagens of various types of this protein does not differ significantly either, whereas solubility of the collagen in aneurysmal wall and its susceptibility to the action of EDTA are distinctly decreased. In contrast with collagen, the amount of elastin in aneurysms is significantly lower. Total amount of glycosaminoglycans slightly decreased as compared with that of normal tissue, but the ratio of particular compounds varies. The percentage of chondroitin sulphate is increased and that of heparan sulphate significantly decreased. The significance of these changes in pathogenesis of aneurysms is discussed.

Topics: Adult; Aged; Aging; Aortic Aneurysm, Abdominal; Arteriosclerosis; Case-Control Studies; Chondroitin Sulfates; Collagen; Elastin; Female; Glycosaminoglycans; Humans; Keratan Sulfate; Male; Middle Aged; Solubility

We recently isolated a proteoglycan form of macrophage colony-stimulating factor (PG-M-CSF) that carries a chondroitin sulfate glycosaminoglycan chain. Here, we examined the interaction of PG-M-CSF with low density lipoprotein (LDL). When LDL preincubated with PG-M-CSF was fractionated by molecular size sieving chromatography, it was eluted earlier than untreated LDL. When LDL was preincubated with chondroitin sulfate-free 85-kD M-CSF instead of PG-M-CSF, the elution profile of LDL remained unchanged, indicating specific interaction between PG-M-CSF and LDL. The level of PG-M-CSF binding in the wells of a plastic microtitration plate precoated with LDL was significant, this binding being completely abolished by pretreatment of PG-M-CSF with chondroitinase AC, which degrades chondroitin sulfate. The addition of exogenous chondroitin sulfate or apolipoprotein B inhibited the binding of PG-M-CSF to LDL in a dose-dependent manner, indicating that the interaction between PG-M-CSF and LDL was mediated by the binding of the chondroitin sulfate chain of PG-M-CSF to LDL apolipoprotein B. PG-M-CSF was also demonstrated in the arterial wall, and there were increased amounts of PG-M-CSF in atherosclerotic lesions. The in vitro interaction between PG-M-CSF and LDL thus appears to have physiological significance.

Topics: Aorta; Apolipoproteins B; Arteriosclerosis; Chondroitin Sulfates; Humans; Lipoproteins, LDL; Macrophage Colony-Stimulating Factor; Protein Binding

The topographic distribution of atherosclerotic lesions is influenced by biochemical factors intrinsic to the arterial wall. In the present work we have investigated whether the composition/chemical structure of glycosaminoglycans constitutes one of these factors. Normal human arteries were obtained at necropsy, and in order of decreasing susceptibility to atherosclerosis, consisted of the abdominal and thoracic aortas and the iliac and pulmonary arteries. The results showed similar concentrations of total glycosaminoglycan and collagen. Of the glycosaminoglycans known to interact with low-density lipoprotein (LDL), dermatan sulfate was present in all arteries in comparable concentrations, but the aortas had a 30% higher content of chondroitin 4/6-sulfate, which in turn was slightly enriched in 6-sulfated disaccharide units. LDL-affinity chromatography with dermatan sulfate+chondroitin 4/6-sulfate fractions demonstrated that increasing affinity to LDL matched an increasing susceptibility to atherosclerosis. Analysis of glycosaminoglycans in the eluates indicated a positive correlation between affinity to LDL and increasing molecular weight and the existence of a fraction of glycosaminoglycans of high affinity to LDL in the aortas only. These results suggest that arterial glycosaminoglycans participate in the multifactorial mechanisms that modulate the differential localization of atherosclerotic lesions.

Topics: Adult; Aorta, Abdominal; Aorta, Thoracic; Arteries; Arteriosclerosis; Chondroitin Sulfates; Chromatography, Affinity; Dermatan Sulfate; Glycosaminoglycans; Humans; Iliac Artery; Lipoproteins, LDL; Pulmonary Artery

A chondroitin sulfate-dermatan sulfate proteoglycan was isolated from bovine aorta intima by extraction of the tissue with 4 M guanidine hydrochloride. The proteoglycan was purified by CsCl isopycnic centrifugation followed by gel filtration and ion exchange chromatography. A monoclonal antibody C8F4 was developed to this core protein. The characteristics and specificity of the antibody were studied by an enzyme-linked immunosorbent assay (ELISA) using an alkaline phosphatase conjugated antibody (goat anti-mouse IgG). The antibody binding to the core protein was found specific and optimal at pH 7.0. The antibody recognizes either intact chondroitin sulfate-dermatan sulfate proteoglycan monomer, chondroitinase ABC digested monomer or chemically deglycosylated proteoglycan. Free chondroitin sulfates, keratan sulfate and hyaluronic acid did not compete for the antigenic sites in ELISA. Limited hydrolysis of the core protein by trypsin resulted in three peptides and only the peptide with a molecular weight M(r) = 40,000 was found capable of binding to hyaluronic acid. The antibody C8F4 recognized this hyaluronic acid binding peptide but did not recognize the other two peptides suggesting that the epitope(s) for this antibody is in the hyaluronic acid-binding region of the core protein. The antibody recognized the core proteins from bovine nasal cartilage proteoglycan and human aorta proteoglycan but did not recognize bovine aorta link protein, bovine serum albumin, human serum albumin, human transferrin, collagen Type I and fibronectin. The antibody was found useful to localize proteoglycans in atherosclerotic lesions in human aorta by immunohistochemical techniques.

Topics: Animals; Antibodies, Monoclonal; Antibody Specificity; Aorta; Arteriosclerosis; Binding Sites; Cattle; Chondroitin Sulfates; Enzyme-Linked Immunosorbent Assay; Epitopes; Humans; Hyaluronic Acid; Mice; Mice, Inbred BALB C

This study was designed to clarify the effects of vitamin E on the alterations in proteoglycan distribution and vascular permeability, which were examined by immunohistochemical and ultrastructural techniques in the aortas of cholesterol-fed guinea pigs. The animals were divided into three groups: a control group, a cholesterol group, and a vitamin E group. Serum levels of cholesterol, triglyceride, low-density lipoprotein, high-density lipoprotein, and thiobarbituric acid-reactive substances were measured. An increase in thiobarbituric acid-reactive substances was observed in the cholesterol group compared with control and vitamin E groups. Intimal atheromatous lesions of the aorta were significantly decreased in the vitamin E group compared with the cholesterol group. Histochemically, an increased distribution of proteoglycans such as chondroitin, dermatan, and heparan sulfates and ruthenium red reaction products in the intima; decreased glycocalyx on the endothelial surface; and increased permeability to horseradish peroxidase were revealed in the cholesterol group compared with the vitamin E group. Hypercholesterolemia, resulting in superoxide production, may have contributed to the endothelial damage and increased permeability to plasma proteins and lipids in the vascular wall of the cholesterol group. However, vitamin E administration inhibited lipid deposition and development of this abnormal permeability associated with an irregular distribution of proteoglycan. These results suggest that vitamin E preserves the morphological and functional integrity of the vascular wall and may contribute to the inhibition of atherogenesis in cholesterol-fed guinea pigs.

Topics: Animals; Aorta; Arteriosclerosis; Capillary Permeability; Cholesterol; Cholesterol, Dietary; Chondroitin Sulfates; Dermatan Sulfate; Guinea Pigs; Heparitin Sulfate; Immunohistochemistry; Lipoproteins; Microscopy, Electron; Thiobarbituric Acid Reactive Substances; Vitamin E

We have shown recently that lipoprotein-proteoglycan complexes isolated from human atherosclerotic lesions stimulated cholesteryl ester synthesis in human monocyte-derived macrophages [Vijayagopal, Srinivasan, Radhakrishnamurthy and Berenson (1992) Arterioscler. Thromb. 12, 237-249]. The present study was conducted to determine the mechanism of cellular uptake of the complexes. A chondroitin sulphate-dermatan sulphate proteoglycan was isolated from normal human aorta and complexed to 125I-labelled human low-density lipoprotein (LDL). The binding and degradation of 125I-LDL-proteoglycan complex were then studied in human monocyte-derived macrophages. The specific binding and degradation of the complex showed saturability and concentration-dependency. The Kd for binding was 1.5 x 10(-8) M, which was greater than that reported for LDL in monocyte-derived macrophages. Binding of the complex was not subject to down-regulation. Chloroquine inhibited degradation of the complex and the resultant stimulation of cholesteryl ester synthesis. Limited treatment of macrophages with proteolytic enzymes abolished binding and degradation of the complex significantly. Macrophages bound 125I-methyl-LDL-proteoglycan complex to the same extent as 125I-LDL-proteoglycan complex. Excess LDL and proteoglycan did not compete against the binding of the complex; however, excess acetyl-LDL competed for 61% of the binding. Likewise, excess LDL-proteoglycan complex inhibited the binding of 125I-acetyl-LDL by 64%. Polyinosinic acid and cytochalasin D inhibited the binding of 125I-LDL-proteoglycan complex by 60% and 36% respectively. Compared with that of acetyl-LDL, the degradation of LDL-proteoglycan complex was retarded in human macrophages. The results indicate that the uptake of LDL-proteoglycan complex in human monocyte-derived macrophages is not mediated through binding to the LDL receptor; but occurs predominantly via the scavenger receptor, with phagocytosis playing a minor role in the process.

Topics: Aorta; Arteriosclerosis; Binding, Competitive; Cell Differentiation; Cells, Cultured; Cholesterol Esters; Chondroitin Sulfates; Cytochalasin D; Dermatan Sulfate; Humans; Lipoproteins; Lipoproteins, LDL; Macrophages; Monocytes; Phagocytosis; Proteoglycans; Receptors, Cell Surface; Receptors, Lipoprotein

We studied the metabolism of lipoprotein-proteoglycan complexes by macrophage-derived foam cells. Foam cells were isolated from atherosclerotic rabbit aortas. ApoB-lipoprotein-proteoglycan complex was isolated from human aorta fibrous plaque lesions and LDL-proteoglycan complex was formed in vitro. Both in vitro and in vivo complexes stimulated cholesteryl ester synthesis in foam cells by a dose-dependent, saturable process that resulted in the intracellular accumulation of cholesteryl ester. Stimulation of cholesteryl ester synthesis was linear with time over a 32-h period. Polyinosinic acid inhibited the stimulation of cholesteryl ester synthesis by the complexes by 32-37%, whereas cytochalasin D only produced a 6-16% inhibition. Foam cells degraded 125I-LDL-proteoglycan complex and 125I-acetyl LDL in a saturable, dose-dependent manner. Excess unlabeled acetyl-LDL inhibited the degradation of 125I-LDL-proteoglycan complex by 52%, while LDL had no effect. Similarly, excess unlabeled complex suppressed the degradation of 125I-acetyl-LDL by 48%. Foam cells degraded 125I-methyl-LDL-proteoglycan complex to the same extent as 125I-LDL-proteoglycan complex. These results show that foam cells from atherosclerotic lesions metabolize lipoprotein-proteoglycan complexes predominantly via receptor-mediated endocytosis and consequently continue to accumulate intracellular cholesteryl ester.

Topics: Animals; Aorta; Arteriosclerosis; Cholesterol; Cholesterol Esters; Cholesterol, Dietary; Chondroitin Sulfates; Cytochalasin D; Dermatan Sulfate; Diet, Atherogenic; Foam Cells; Humans; Iliac Artery; Kinetics; Lipoproteins, LDL; Male; Muscle, Smooth, Vascular; Poly I; Proteoglycans; Rabbits

In vitro, chondroitin sulfate (CS) proteoglycans (PGs) bind with high-affinity lipoproteins (LPs) containing apolipoprotein B (apo B), and cultured monocytes incubated with LP-PG complexes transform into foam cells (FCs). Consequently, arterial PGs are thought to contribute to the accumulation of LPs in atherosclerotic lesions, but their in vivo interaction has yet to be demonstrated. Balloon catheterization of rabbit aorta modifies the normal aortic distribution of endogenous LPs containing apo B, and determines their accumulation in normocholesterolemic conditions. The distribution of aortic CS-PGs parallels that of apoB within the neointima of injured aortas and might contribute to a favorable environment for LP sequestration within the lesions. To visualize the in situ relationship between apo B and CS-PG, we performed experiments for double detection by immunofluorescence and by post-embedding electron microscope immunogold. The results indicated that the colocalization of endogenous LPs containing apo B and of the large intimal CS-PGs in advanced lesions developed under the regenerated endothelium. At the ultrastructural level, frequent associations were visualized in the extracellular space and in relation to FCs. The close spatial relation between CS-PG and apo B inside the aortic lesions seems to support the hypothesis of their in situ interaction and of a simultaneous cellular uptake, and may be related to the development of both extracellular lipid deposits and the formation of FCs in atherogenesis.

Topics: Animals; Aorta; Apolipoproteins B; Arteriosclerosis; Chondroitin Sulfates; Endothelium, Vascular; Extracellular Space; Fluorescent Antibody Technique; Microscopy, Electron; Proteoglycans; Rabbits; Tissue Distribution; Wounds, Penetrating

Measurements of total collagen, of the ratio of collagen types III/(I+III) and of sulphated glycosaminoglycans (GAGs) were compared with mechanical strength for individual ulcerated and non-ulcerated human aortic plaque caps and with intima adjacent to the plaques. The distributions of the collagen type ratio were similar for both ulcerated and non-ulcerated plaque caps but different from that of the adjacent intima. The proportions of different collagen types were not related to fracture stress and are thus unlikely to affect the potential to ulcerate. The distributions of the sulphated GAGs showed lower amounts for the plaque caps compared with the nearby intima, with the centres of ulcerated plaque caps having the lowest values. Total collagen had higher values in the peripheries of plaque caps compared with the nearby intima, but was distinctly lower in the centres of ulcerated plaque caps. Plaque caps appeared to require a higher collagen content than adjacent intima to support a given level of mechanical strength, suggesting that while collagen production had occurred in the plaque caps it was not as efficiently organized to resist fracture as a similar amount of collagen in the adjacent intima. Ulcerated plaque caps are notable for much larger transverse (centre vs. periphery) gradients of connective tissue constituents than for non-ulcerated plaque caps. The development of these transverse gradients may be a critical aspect in determining the propensity of a plaque to ulcerate.

Topics: Aortic Diseases; Arteriosclerosis; Chondroitin Sulfates; Collagen; Dermatan Sulfate; Elastin; Glycosaminoglycans; Hexosamines; Humans; In Vitro Techniques; Tensile Strength; Uronic Acids

We have developed a method for using near infrared Raman spectroscopy to quantitatively analyze the histochemical composition of human artery. The main contributors to bands observed in the Raman spectra of normal and atherosclerotic aorta are the proteins collagen and elastin, cholesterol lipids, and calcium hydroxyapatite. The Raman scattering cross-sections of different bands for these components have been determined in order to understand their relative contributions to the Raman spectra of biological tissue. The Raman signal is observed to behave linearly with the concentration of the components, even in a highly scattering medium such as a powder. Using these data, we have developed a linear model that can be used to extract the quantitative contribution of an individual component to the spectrum of a mixture. The model has been applied to several mixtures of known composition of tissue constituents in order to evaluate its precision and accuracy. The calculated fit coefficients from the spectra are in agreement with the measured values within experimental uncertainties. The spectra of different types of atherosclerotic aorta have also been modeled, and we have extracted quantitative information regarding the relative concentration of biological constituents in atherosclerotic aorta.

Topics: Arteries; Arteriosclerosis; Cholesterol; Cholesterol Esters; Chondroitin Sulfates; Collagen; Elastin; Fourier Analysis; Humans; Hyaluronic Acid; Muscle, Smooth, Vascular; Palmitic Acid; Palmitic Acids; Spectrophotometry, Infrared; Spectrum Analysis, Raman; Triglycerides

The contents of three species of proteoglycans (PGs), heparan sulfate PG(HSPG), chondroitin sulfate PG(CSPG) and dermatan sulfate chondroitin sulfate PG(DSCSPG), in human thoracic aortas of subjects from districts of high (Beijin, in North China) and low (Nanning, in South China) prevalence of atherosclerosis in China were quantitated. Higher aortic HSPG and DSCSPG (but lower DS) in samples from Nanning than those from Beijing might be implicated in the lower prevalence of atherosclerosis in the former.

Topics: Adolescent; Adult; Aorta, Thoracic; Arteriosclerosis; China; Chondroitin Sulfate Proteoglycans; Chondroitin Sulfates; Chromatography, Gel; Chromatography, Ion Exchange; Dermatan Sulfate; Electrophoresis, Cellulose Acetate; Female; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Male; Prevalence; Proteoglycans

Proteoglycan (PG) metabolism by aortic smooth muscle cell cultures derived from atherosclerosis-susceptible White Carneau (WC) and -resistant Show Racer (SR) pigeons was compared using [35S]sodium sulfate and [3H]serine or [3H]glucosamine as labeling precursors. Chondroitin sulfate (CS) PG and dermatan sulfate (DS) PG were the major PG secreted into the medium by both cell types. Total PG production, whether measured by incorporation of radiolabel into either core protein or glycosaminoglycan chains, was consistently lower in WC compared to SR cultures at several time points. This difference was due in part to lower (30-37%) PG synthesis in WC cells, but degradation of newly synthesized PG was an important contributor. A pulse-chase study indicated that of the total radiolabeled PG present at time O, only 47% was present at 24 h in WC cultures compared to 88% in SR cultures. The large CS-PG appeared to be the primary target for degradation in WC cells, and this selective processing resulted in a higher DS-PG:CS-PG ratio in these cultures. Structural studies indicated similar core protein and glycosaminoglycan chain sizes within a PG type for both cell types. PG monomer composition differed, however, by a higher sulfation of WC CS-PG compared to SR CS-PG and by a disaccharide sulfation position favoring 6-sulfation in WC PG and 4-sulfation in SR PG.

Topics: Animals; Aorta; Arteriosclerosis; Cells, Cultured; Chemical Precipitation; Chondroitin Sulfates; Chromatography; Columbidae; Culture Media; Dermatan Sulfate; Disaccharides; Glucosamine; Glycosaminoglycans; Kinetics; Muscle, Smooth, Vascular; Proteoglycans; Serine; Sulfates

We compared the effects of mild hypercholesterolemia and repeated endotoxin infusions on the biochemical composition of aortic intima and inner media of 24 piglets divided into 4 groups 5 days after weaning: controls on normal diet (group I); normal diet and endotoxin (group II); fat-supplemented diet (group III); and fat-supplemented diet and endotoxin (group IV). It was found that mild hypercholesterolemia increased the concentration of arterial esterified cholesterol and the relative amount of the fraction containing chondroitin sulphates A and C in total glycosaminoglycans. Endotoxin infusions partly prevented the increase of serum cholesterol caused by the fat-supplemented diet but had no independent effect on the arterial biochemical composition; nor did they affect the biochemical changes caused by hypercholesterolemia. When the results of all groups were combined, chondroitin sulphates A and C showed a significant positive correlation with the concentration of arterial esterified cholesterol and the percentage of linoleic acid in arterial cholesteryl esters. Serum total cholesterol did not correlate with arterial cholesterol fractions, but the ratio of high density lipoprotein-cholesterol to total serum cholesterol showed a negative association with arterial esterified cholesterol. The present findings indicate that (1) mild hypercholesterolemia is atherogenic in young piglets, and (2) changes in arterial glycosaminoglycan composition might be one of the earliest biochemical alterations in atherogenesis.

Topics: Animals; Aorta, Thoracic; Arteriosclerosis; Cholesterol; Chondroitin Sulfates; Dermatan Sulfate; Diet, Atherogenic; Endotoxins; Glycosaminoglycans; Heparitin Sulfate; Hyaluronic Acid; Hypercholesterolemia; Lipids; Muscle, Smooth, Vascular; Phospholipids; Reference Values; Swine

Artery proteoglycan-lipoprotein binding characteristics were determined using intact, high molecular weight chondroitin sulfate proteoglycans (CS-PG) isolated from grossly appearing normal aortas of atherosclerosis susceptible WC-2 pigeons and plasma lipoproteins from normolipemic, randomly bred White Carneau pigeons. Optimum formation of particulate proteoglycan-lipoprotein complexes occurred in 5 mM Tris, 6 mM KCl, 4 mM CaCl2, 1 mM MgSO4, pH 7.2. The binding of CS-PG was specific for low density lipoprotein (LDL) and not high density lipoprotein (HDL). The relative importance of the intact monomeric structure of the PG was suggested in studies where glycosaminoglycan chains isolated from the PG monomer possessed less than 1% of the binding reactivity of the intact PG. The core protein prepared from the CS-PG monomer formed no measurable particulate complex.

Topics: Animals; Arteries; Arteriosclerosis; Chondroitin Sulfates; Columbidae; Dose-Response Relationship, Drug; Electrophoresis; Female; Hydrogen-Ion Concentration; Ions; Lipoproteins; Lipoproteins, LDL; Male; Protein Binding; Protein Conformation; Proteoglycans; Temperature

The binding of intact, high molecular weight, aortic proteoglycan (PG) isolated from grossly normal appearing aortas of atherosclerosis-susceptible White Carneau pigeons (WC-2) and -resistant Show Racer pigeons (SR) to homologous and heterologous serum lipoproteins from both normolipemic and hyperlipemic pigeons was examined. In vitro binding studies were done using a mixture of purified chondroitin sulfate PG and dermatan sulfate PG monomers to simulate an in situ composition. For each animal, a binding potential or reactivity number was calculated and corresponded to the shape and slope of the PG-LDL binding curve, where higher values indicated greater reactivity. For WC-2 normal sera, mean values were 0.97 and 0.95 using WC-2 and SR PG respectively, compared to SR normal sera (equivalent total plasma cholesterol) where values were 0.80 and 0.82. Corresponding mean reactivity values for hyperlipemic sera (diluted to a cholesterol concentration of 300 mg/dl) were 0.95, 1.00, 0.73, and 0.79. The results suggest that LDL from both normolipemic and hyperlipemic atherosclerosis-susceptible WC-2 pigeons is more reactive in complexing to artery wall derived PG than LDL from SR pigeons, regardless of PG source.

Topics: Animals; Aorta, Thoracic; Arteriosclerosis; Chondroitin Sulfates; Columbidae; Disease Susceptibility; Electrophoresis, Agar Gel; Hyperlipidemias; Immunity, Innate; Lipoproteins; Lipoproteins, LDL; Protein Binding; Protein Conformation; Proteoglycans

The effect of chondroitin-6-sulfate, obtained from shark cartilage, on atherogenesis in rabbits fed a high-cholesterol diet was studied. Male Japanese white rabbits were housed for 10 weeks in three groups, one group was fed ordinary pellets and was injected intraperitoneally with saline (standard-diet group), one was fed pellets containing 1% cholesterol and was injected intraperitoneally with saline (cholesterol-diet group), and the third group was fed pellets containing 1% cholesterol, and was injected intraperitoneally with 10 mg of chondroitin-6-sulfate (C-6-S group). Injections were done daily. The plasma total cholesterol, and cholesterol from very low-density lipoprotein in the C-6-S group after 5 weeks in the test period, and low-density lipoprotein cholesterol in the C-6-S group at the end of the test period were lower than those of the cholesterol-diet group. Significantly fewer atherosclerotic lesions of the aortic surface were found macroscopically in the C-6-S group than in the cholesterol-diet group. The cholesterol, esterified cholesterol and calcium concentrations of the aortic intima-media in the C-6-S group were significantly lower than in the cholesterol-diet group. Hydroxyproline levels in these three groups were not different. The uronic acid concentration of the intima-media in the cholesterol-diet group was significantly higher than in the C-6-S group (P less than 0.02). Though the percentage of heparan sulfate on total glycosaminoglycans (GAGs) of the C-6-S group was lower than in the cholesterol-diet group, there were no significant differences in the percentages of dermatan sulfate and chondroitin-4/6-sulfate in total GAGs between the cholesterol-diet and C-6-S groups. These results suggest that chondroitin-6-sulfate suppresses cholesterol deposition in the aorta of rabbits fed a 1% cholesterol diet, probably partly due to a decrease in the plasma low-density lipoprotein cholesterol, and partly due to a change in arterial metabolism.

Topics: Animals; Aorta; Arteriosclerosis; Calcium; Chondroitin; Chondroitin Sulfates; Glycosaminoglycans; Heparin; Hydroxyproline; Hypercholesterolemia; Lipid Metabolism; Lipolysis; Lipoproteins; Male; Platelet Aggregation; Rabbits

Male Dutch-belted rabbits were fed a pelleted 0.5% cholesterol diet for 8 weeks. A control group was maintained on basal pelleted diet for the same period of time. Prior to sacrifice both groups received i.v. 3H-inulin. Plasma and aortic sections were assayed for 3H-inulin activity. Aortic sections were also analyzed for glycosaminoglycan concentrations. A ratio of the aortic 3H-inulin activity to the plasma 3H-inulin activity was calculated and expressed as a permeability index. Mean plasma activity of 3H-inulin was significantly lower in the control group, suggesting increased aortic permeability in the cholesterol-fed animals. Glycosaminoglycan concentrations were significantly higher in the control group, and significant negative correlations were detected between glycosaminoglycan concentrations and permeability index in all animals. These findings are consistent with the hypothesis that sufficient glycosaminoglycan levels are important to maintaining the permeability characteristics of the endothelial layer.

Topics: Animals; Aorta, Thoracic; Arteries; Arteriosclerosis; Cholesterol, Dietary; Chondroitin Sulfates; Diet, Atherogenic; Endothelium; Glycosaminoglycans; Hypercholesterolemia; Inulin; Male; Permeability; Rabbits

Chondroitin sulphates (CSs) were isolated from the intima and the media of normal (normal CSs) and atherosclerotic (sclerotic CSs) regions of human aortas. Normal and sclerotic CSs accelerated the inactivation of thrombin by antithrombin III to an equal extent. By this mechanism, both normal and sclerotic CSs prolonged thrombus formation time in a moving stream of platelet-rich plasma and thrombin-catalysed clotting time of platelet-poor plasma. However, anticoagulant activity of sclerotic CSs in thrombin-catalysed fibrin clot formation in platelet-poor plasma was lower than that of normal CSs. The lower anticoagulant activity of sclerotic CSs was due to the greater accelerating effect on the polymerization of monomeric fibrin to form clot, which was the final distinguishable step of fibrinogen-fibrin conversion.

Topics: Adult; Anticoagulants; Aorta, Thoracic; Arteriosclerosis; Blood Coagulation; Chondroitin; Chondroitin Sulfates; Dose-Response Relationship, Drug; Fibrin; Humans; In Vitro Techniques; Time Factors

Topics: Adolescent; Adult; Age Factors; Aged; Aorta; Arteriosclerosis; Child, Preschool; Chondroitin; Chondroitin Sulfates; Female; Glycosaminoglycans; Humans; Infant; Male; Middle Aged

The composition and content of aortic glycosaminoglycans were studied in groups of rhesus monkeys fed control or atherogenic diets for 9 or 19 months. Aortic uronic acid content was significantly increased in both groups of monkeys with atherosclerosis. The major glycosaminoglycan in both control and atherosclerotic aortas was chondroitin sulfate with lesser amounts of heparan sulfate, dermatan sulfate, and hyaluronic acid. Dermatan sulfate was the only glycosaminoglycan to show a statistically significant elevation (65 to 87 per cent) in animals fed the atherogenic diet. This increase was positively correlated with the increased accumulation of aortic cholesterol (r = 0.4709, p less than 0.05). The results indicate that dermatan sulfate may be the major glycosaminoglycan involved during the early events of atherogenesis perhaps through retention of lipoprotein in the atherosclerotic artery.

Topics: Animals; Aorta; Arteriosclerosis; Cholesterol; Chondroitin; Chondroitin Sulfates; Dermatan Sulfate; Diet, Atherogenic; Glycosaminoglycans; Haplorhini; Macaca mulatta; Male; Uronic Acids

Topics: Aorta; Arteriosclerosis; Chondroitin Sulfates; Factor VIII; Fibrin; Glycosaminoglycans; Heparin; Humans; Polymers; Solubility; Thrombelastography; Thrombosis; Tissue Extracts; Trypsin

Topics: Aged; Arteriosclerosis; Cholesterol; Chondroitin; Chondroitin Sulfates; Female; Humans; Male; Placebos; Triglycerides

Heparin-like glycosaminoglycans (GAG) were isolated from commerical Vessel and their biologic properties studied. Vessel was found to be a mixture of chondroitin sulfates, dermatan sulfate and heparin-like GAG. Chondroitin sulfates and dermatan sulfate in Vessel were hydrolyzed by chondroitinase ABC and the residual Vessel was fractionated on a Dowex-1 Cl- column eluting with a stepwise-increasing concentration of NaCl (1.2--4.0 M). The major fractions eluted at 1.6 M and 1.8 M NaCl were tentatively identified by chemical analysis as heparin-like GAG with somewhat lower sulfate content than standard heparin. Both fractions had lipoprotein lipase-releasing activity and anticoagulant activity similar to heparin, but 1.6 M NaCl fraction had a third of the anticoagulant activity of standard heparin. The 1.8 M NaCl fraction complexed with serum lipoproteins similarly to heparin. In preliminary studies cholesterol-fed rabbits treated with Vessel exhibited somewhat less atherosclerosis than controls.

Topics: Animals; Arteriosclerosis; Cholesterol, Dietary; Chondroitin Sulfates; Glycosaminoglycans; Hypolipidemic Agents; Lipoprotein Lipase; Lipoproteins; Rabbits

Topics: Aging; Animals; Aorta, Thoracic; Arteriosclerosis; Chondroitin Sulfates; Columbidae; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate; Hyaluronic Acid; Species Specificity

Topics: Animals; Arteriosclerosis; Carbamates; Cholesterol; Chondroitin; Chondroitin Sulfates; Cricetinae; Guinea Pigs; Hypolipidemic Agents; Liver; Mice; Pentanoic Acids; Pyrazoles; Pyridinolcarbamate; Rabbits; Rats; Valerates

The changes in levels of glycosaminoglycans (GAGs) of the intima and media of the human artery in atherosclerosis were determined by a recently introduced two-dimensional electrophoresis technique that permits direct measurments of each of these macromolecules. To identify the arterial GAGs, they were fractionated by chromatography on a DEAE-Sephadex A-25 column, and the resulting three fractions (hyaluronic acid [HA], heparan sulfate [HS], and the partially separated chondroitin sulfates B [CSB] and C [CSC]) were analyzed for their electrophoretic mobilities by this electrophoretic method, for their digestability by highly specific hydrolases (leech hyaluronidase, heparinase, and chondroitinases ABC and AC) and for their iduronic acid content. From these studies we concluded that normal and atherosclerotic human aortas contain CSB, CSC, HA, and HS. Further, we demonstrated that CSB is a hybrid consisting of approximately 40% CSA and 60% CSB and that CSC appears to be a polymer consisting essentially of glucuronic acid and N-acetylgalactosamine-6-sulfate. Classical CSA as well as chondroitin (CH) were not present in detectable amounts. In the relatively normal intima, the mean concentrations of the GAGs were found to be 4.7, 20.9, 1.3, and 5.1 mg/g of dry, defatted, decalcified tissue for CSB, CSC, HA, and HS, respectively. With the progression of atherosclerosis, there was a pronounced decrease in the total GAG content (from 32 to 18 mg) associated with a decrease in the CSC and HS levels but without a change in the HA concentrations. Of particular interest, however, was the increase in the CSB level. In the media whose total GAG content averaged approximately 20 mg, no significant changes in these GAG levels were noted with the progression of the disease except for that of CSC. These findings may be important in explaining the increased lipoprotein and collagen deposition in the diseased aorta.

Topics: Aorta; Arteries; Arteriosclerosis; Chondroitin Sulfates; Electrophoresis, Cellulose Acetate; Glycosaminoglycans; Humans; Hyaluronic Acid

Fifteen chicken tissues were examined for mast cells. Numerous mast cells were found in peritoneum and spleen. The sulfated mucopolysaccharides extracted from these two tissues, corresponding in amount to that in mast cells, were found to be dermatan sulfate, chondroitin sulfate and heparitin sulfate but no evidence of heparin. We have shown a similar situation occurs in the rabbit which is also highly susceptible to the production of atherosclerosis by diet. These observations provide further evidence of a role for heparin and mast cells in limiting atherogenesis.

Topics: Animals; Arteriosclerosis; Cell Count; Chickens; Chondroitin Sulfates; Dermatan Sulfate; Dogs; Female; Heparin; Heparitin Sulfate; Male; Mast Cells; Peritoneum; Rats; Spleen

Topics: Animals; Arteriosclerosis; Carbamates; Chondroitin; Chondroitin Sulfates; Mevalonic Acid; Pyridinolcarbamate; Rabbits

Topics: Arteriosclerosis; Blood Coagulation; Cholesterol; Chondroitin; Chondroitin Sulfates; Hexosamines

Topics: Aorta; Arteriosclerosis; Chondroitin; Chondroitin Sulfates; Chromatography; Nasal Septum; Oligosaccharides; Research; Skin; Sulfates