heparitin-sulfate and Diabetic-Retinopathy

In diabetic nephropathy, heparan sulfate glycosaminoglycan side chains are reduced in glomerular basement membranes proportionally to the degree of proteinuria. Recently, it was demonstrated that additional therapy with danaparoid sodium, a mixture of sulfated glycosaminoglycans with mainly heparan sulfate, lowered proteinuria in type 1 diabetes patients with diabetic nephropathy. A randomized placebo-controlled parallel study was performed with 750 anti-Xa units of danaparoid sodium once daily in type 2 diabetes patients with severe proteinuria. The aim of the study was to evaluate the possible effects of danaparoid sodium on proteinuria, endothelial dysfunction, and hard exudates in the retina and to determine the safety/tolerability of this drug. Twenty-two patients completed the study, and one patient had to stop prematurely after 6 wk of danaparoid sodium treatment because of urticaria at the injection sites. Apart from a small decrease of hemoglobin and minor skin hematomas at the injection site in five patients in the danaparoid sodium group, no other safety parameters showed any clinically or statistically significant difference between and within groups. The relative change in time of both the urinary albumin and protein excretion rate corrected for creatinine did not differ between both treatment arms (P = 0.2 and 0.49, respectively). No retinal complications or changes of hard exudates occurred. von Willebrand factor was elevated in both groups, but was not influenced by either treatment modality. Contrary to the beneficial effects that occurred in type 1 diabetes patients with diabetic nephropathy, treatment for 8 wk with 750 anti-Xa units of danaparoid sodium gave no reduction of proteinuria, hard exudates, and von Willebrand factor.

Topics: Adult; Aged; Analysis of Variance; Anticoagulants; Chondroitin Sulfates; Dermatan Sulfate; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Diabetic Retinopathy; Double-Blind Method; Drug Combinations; Endothelium, Vascular; Exudates and Transudates; Female; Follow-Up Studies; Heparitin Sulfate; Humans; Kidney Function Tests; Male; Middle Aged; Proteinuria; Reference Values; Statistics, Nonparametric; von Willebrand Factor

In diabetes mellitus, both retinopathy and nephropathy represent specific microvascular disease with increased capillary permeability resulting in hard exudates, foveal oedema, and albuminuria. The decrease of heparan-sulphate content of the glomerular-basement membrane is quantitatively related to the rate of proteinuria in nephropathy associated with insulin-dependent diabetes mellitus (IDDM). Several short-term studies in patients with IDDM and non-IDDM have shown that a reduction of microalbuminuria and macroalbuminuria can be achieved with the supplementation of glycosaminoglycans. After completion of a study on the effect of danaparoid sodium on albumin excretion in patients with IDDM and macroalbuminuria, we hypothesised that treatment with danaparoid sodium also influenced retinal leakage in the patients in that trial.. In this retrospective study nine patients with nephropathy received 750 anti-Xa units danaparoid sodium once a day for 6 weeks in a placebo-controlled double-blind cross-over study. Fundus photographs, done at baseline and at the end of the study, were semiquantitatively scored for the severity of hard exudates.. At baseline 14 eyes had grade 1 to 5 severity of hard exudates and four eyes were without hard exudates (grade 0). There was no progression in the latter four eyes. In ten eyes an improvement was observed: four patients showed a favourable response to treatment in both eyes and two patients showed improvement in one eye. We found improvement of hard exudates after 6 weeks of treatment with danaparoid sodium.. Our uncontrolled observation indicates that the supplementation of danaparoid sodium influences both the permeability of retinal vessels as well as of glomerular vessels. Danaparoid-sodium therapy as a systemic adjuvant is worth considering for treatment strategies for foveal oedema and hard exudates in diabetic maculopathy.

Topics: Adult; Chondroitin Sulfates; Cross-Over Studies; Dermatan Sulfate; Diabetic Retinopathy; Double-Blind Method; Drug Combinations; Exudates and Transudates; Female; Heparitin Sulfate; Humans; Male; Retrospective Studies

To assess the relationship between intraocular soluble heparan sulfate (HS) concentration and age in subjects with and without diabetic retinopathy.. Vitreous from subjects with idiopathic maculopathies (n=17), i.e., macula hole or epiretinal membrane, or nonproliferative diabetic retinopathy (non-PDR; n=5) and aqueous humor from subjects with PDR (n=16), non-PDR (n=7), or cataracts (n=15) was collected. The levels of HS and vascular endothelial growth factor (VEGF) were measured using enzyme-linked immunosorbent assay. Concentrations of sulfated glycosaminoglycan were determined through dimethylmethylene blue-based assay. The effect of the vitreal HS level on the binding of exogenous VEGF to surface-bound heparin was determined in vitro.. The level of HS in vitreous samples from subjects with idiopathic maculopathies increased concomitantly with age (p=0.020, R²=0.327). Meanwhile, HS levels in aqueous humor were lower in PDR subjects than in non-PDR (p=0.003) and cataract subjects (p=0.007). However, the PDR subjects were significantly younger than the non-PDR subjects (p<0.001) or cataract subjects (p<0.001). When the three groups were controlled for age, the levels of HS glycosaminoglycans were no longer different between the three (p=0.247). The increasing level of HS or sulfated glycosaminoglycan in the vitreous was associated with its increased inhibitory effect on interaction between VEGF and surface heparin in vitro (p=0.014, R²=0.377).. The HS level of the intraocular fluid increased with age. The possible link between low HS in intraocular fluid and increased localization of VEGF at the retinal surface may provide one explanation for the higher susceptibility of younger subjects with diabetes mellitus to developing PDR.

Topics: Adult; Aged; Aged, 80 and over; Aging; Aqueous Humor; Diabetic Retinopathy; Heparitin Sulfate; Humans; Middle Aged; Protein Binding; Solubility; Vascular Endothelial Growth Factor A; Vitreous Body; Young Adult

The Steno hypothesis (Deckert et al. ) states that in diabetes mellitus (DM), changes in vascular heparan sulfate proteoglycan (HSPG) expression are involved in systemic endothelial dysfunction and increased capillary permeability. In diabetes-induced glomerular capillary leakage, loss of HSPG and its side chains has been documented. This study aimed to investigate whether microvascular leakage in diabetic retinopathy (DR) is also associated with altered expression of HSPG in retinal microvessels.. Serial cryosections of post-mortem eyes of 22 subjects with DM and 7 controls were stained with antibodies against the core proteins of the basement membrane HSPGs agrin (Abs Bl31 and JM72) and perlecan (Ab 1948), and four antibodies against heparan sulfate side chains (HS) (Abs JM403, HepSS1, JM13, 3G10). Moreover, we investigated Cynomolgus monkey eyes injected with vascular endothelial growth factor (VEGF)-A, as a model of retinal microvas-cular leakage. The endothelial antigen PAL-E was used to detect microvascular leakage.. In the retina of all controls and DM cases, agrin and perlecan core proteins and HS as recognized by JM403 and 3G10 were expressed in the walls of microvessels. Staining for JM13 was variable between cases, but unrelated to microvascular leakage as determined by PAL-E. Staining for HepSS1 was absent in all human retinal microvessels. In monkey retinas, HSPG staining was identical to that in human retinal tissues, except for the staining for HepSS1, which was found absent in control monkey eyes but which was positive in VEGF-injected eyes.. Increased microvascular permeability in human DR is not associated with changes in expression of the HSPGs studied, whereas high amounts of VEGF may induce increased expression of the HS side chain epitope recognized by HepSS1. These results suggest that the mechanism underlying retinal leakage is different from diabetic glomerular capillary leakage.

Topics: Aged; Aged, 80 and over; Agrin; Animals; Antibody Specificity; Blood-Retinal Barrier; Capillary Permeability; Diabetes Mellitus; Diabetic Retinopathy; Disease Models, Animal; Endothelial Growth Factors; Fluorescent Antibody Technique, Indirect; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Immunoenzyme Techniques; Macaca fascicularis; Middle Aged; Retinal Vessels; Vascular Endothelial Growth Factor A

The development of diabetic nephropathy shows remarkable variation among individuals. Therefore, not only hyperglycemia but also genetic factors may contribute to the development of diabetic nephropathy Heparan sulfate proteoglycan (HSPG) is thought to play an important role as a component of the charge selectivity barrier in the glomerular basement membrane. Recently, a BamHI restriction fragment length polymorphism (RFLP) in the HSPG gene (HSPG2) was reported to be associated with diabetic nephropathy in Caucasian insulin-dependent diabetes mellitus (IDDM). The aim of the present study was to examine the contribution of the BamHI HSPG2 polymorphism to the development of diabetic nephropathy in Japanese non-insulin-dependent diabetes mellitus (NIDDM). For this purpose, we recruited 102 patients with diabetic nephropathy and 64 age-matched patients without diabetic nephropathy from Japanese NIDDM patients. Since all the subjects had proliferative diabetic retinopathy, it seems likely that they would be exposed to hyperglycemia for a long time. In the present study, the BamHI HSPG2 genotype and allele frequencies were not significantly different between the patients with nephropathy and the patients without nephropathy. Therefore, we conclude that the BamHI HSPG2 polymorphism is not associated with the development of diabetic nephropathy in Japanese NIDDM.

Topics: Case-Control Studies; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Diabetic Retinopathy; Female; Heparitin Sulfate; Humans; Japan; Male; Middle Aged; Polymorphism, Genetic; Polymorphism, Restriction Fragment Length; Proteoglycans

Topics: Chondroitin Sulfates; Dermatan Sulfate; Diabetic Retinopathy; Drug Combinations; Endothelial Growth Factors; Heparinoids; Heparitin Sulfate; Humans; Lymphokines; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Basic fibroblast growth factor (FGF) is a potent endothelial cell mitogen that has been proposed to play a role in proliferative diabetic retinopathy and other neovascular processes. Our understanding of the in vivo role of basic FGF in the pathogenesis of these disorders is limited. We studied the immunolocalization of basic FGF in 16 clinical cases of diabetic retinopathy to determine whether the normal retinal distribution of basic FGF changed during the development of diabetic retinopathy and correlated with the onset of retinal neovascularization. By using monoclonal and affinity-purified polyclonal antibodies against basic FGF and heparan sulfate proteoglycan (HSPG), we found that basic FGF colocalized with HSPG to vascular basement membranes. As the basement membranes thickened during the progression of diabetic retinopathy, the intraretinal stores of immunoreactive basic FGF and HSPG expanded. With the development of neovascularization, the colocalization of basic FGF and HSPG changed; HSPG localized to basement membranes, while basic FGF localized intracellularly, with only minimal basement membrane immunoreactivity. Incubations of the neovascular fronds with exogenous basic FGF demonstrated multiple HSPG glycosaminoglycan-binding sites for basic FGF, indicating that basic FGF had not been released from the matrix of neovascular fronds by heparitanase digestion.

Topics: Antibodies, Monoclonal; Basement Membrane; Chondroitin Sulfate Proteoglycans; Diabetic Retinopathy; Fibroblast Growth Factors; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Immunoenzyme Techniques; Polysaccharide-Lyases; Retina; Retinal Neovascularization

Topics: Blood Coagulation; Chondroitin Sulfate Proteoglycans; Diabetes Mellitus, Type 1; Diabetic Retinopathy; Fibrinopeptide A; Glycosaminoglycans; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Proteoglycans