alcian-blue and Glomerulonephritis

Some preliminary observations suggest that predisposition to a particular type of glomerulonephritis (GN) may be connected with the genetically determined charge of the glomerular capillary wall. A correlation between erythrocyte surface and the glomerular capillary wall charges has also been observed. The purpose of this study was to verify and extend previous investigations. Therefore we measured erythrocyte and platelet surface charge from patients with idiopathic membranous and mesangial GN as well as idiopathic membranoproliferative GN and lupus nephritis.. The erythrocyte and platelet surface charge was determined by the binding of the cationic dye, alcian blue (AB). A fresh alcoholic AB solution was made for each experiment, which were run in batches of four, each including cells from a healthy person and from patients each with a different type of GN.. In patients with idiopathic membranous and membranoproliferative GN, a significant decrease in the erythrocyte and platelet charges was observed irrespective of their clinical state (remission or nephrotic syndrome). Erythrocyte charge was decreased despite the normal amount of membranous sialic acid. In contrast, patients with idiopathic mesangial GN, in complete or partial remission, exhibited normal erythrocyte and platelet surface charges. Exclusively in this type of GN, the appearance of nephrotic proteinuria was associated with a slight decrease, the erythrocyte charge, which was not statistically significant (P > 0.1). A reduction in the negative erythrocyte charge in lupus nephritis was less in magnitude than in idiopathic membranous or membranoproliferative GN, and occurred independently of the level of daily proteinuria, whereas the platelet charge was normal.. The decrease of the erythrocyte and platelet charge in idiopathic membranous and mebranoproliferative GN seems to be a pre-morbid feature.

Topics: Adolescent; Adult; Aged; Alcian Blue; Blood Platelets; Case-Control Studies; Cell Membrane; Coloring Agents; Electrochemistry; Erythrocyte Membrane; Female; Glomerulonephritis; Glomerulonephritis, Membranoproliferative; Glomerulonephritis, Membranous; Humans; Lupus Nephritis; Male; Membrane Potentials; Middle Aged; Nephrotic Syndrome; Proteinuria; Surface Properties

We studied alcian blue (AB) binding to red blood cells (RBC) [ABRBC] in 89 children and adults with a variety of glomerular diseases. ABRBC was significantly reduced in the group as a whole when compared with healthy controls (P < 0.001). A moderate correlation between the degree of proteinuria and ABRBC was detected in the children with renal disease (r = 0.43, P < 0.001), but not in adult patients. A significant reduction in ABRBC was detected in 30 of the above children who had Steroid Responsive Nephrotic Syndrome (MCNS) compared to their controls (40.57 +/- 16.6 v. 75.09 +/- 7.81, P = 0.001). A similar decrease from normal values was observed in other childhood glomerular diseases: focal glomerulosclerosis, post-streptococcal glomerulonephritis, membranous nephropathy (52.31 +/- 26.07 v. 79.44 +/- 5.88, P = 0.001). Mean ABRBC was lower in MCNS than other histological groups (P = 0.0518). ABRBC was age-dependent (r = -0.399, P < 0.02). These findings provide further indirect evidence of the relative roles of charge and size selective filters in glomerular basement membrane (GBM) in renal diseases, and suggest that depletion of anionic charge may be a major cause of proteinuria in MCNS.

Topics: Adolescent; Adult; Alcian Blue; Analysis of Variance; Case-Control Studies; Child; Child, Preschool; Electric Conductivity; Erythrocytes; Female; Glomerulonephritis; Humans; Male; Proteinuria; Regression Analysis; Statistics, Nonparametric

The hallmark of renal diseases involving the glomerulus is the presence of proteinuria. While the routes of pathogenesis of proteinuria have not been established, alterations in the barrier function of the glomerular basement membrane (GBM) have been implicated. We evaluated the effect of streptozotocin diabetes and passive Heymann nephritis (PHN) over time on the macromolecular composition of rat GBM to determine if changes in composition correlate with proteinuria. Six to twelve rats from each group (control, diabetic, and PHN) were sacrificed 1, 5, 28, 56, or 84 days after induction of disease. Identical amounts of GBM were subjected to a sequential extraction procedure, and type IV collagen, entactin, laminin, fibronectin, and anionic charge content were quantitated in the extracts. Type IV collagen and entactin content did not change with time or disease. Both laminin and fibronectin contents increased with time in GBM in all groups, but this increase was significantly greater in diabetic GBM. A significant decrease in anionic charge content of GBM coincided with the onset of albuminuria at Day 28 in diabetes, but no change was seen in PHN. In diabetic rats, the increase in laminin content over control preceded the onset of albuminuria, while the increase in fibronectin was not apparent until after albuminuria was present. In PHN, no differences in type IV collagen, entactin, laminin, fibronectin, or anionic charge content of GBM were found compared with control, even though profound albuminuria was evident from Day 5 through 84. Thus, while alterations in laminin and fibronectin content may contribute to the loss of glomerular permselectivity in streptozotocin diabetes, such changes apparently are not involved in PHN.

Topics: Albuminuria; Alcian Blue; Animals; Anions; Basement Membrane; Collagen; Coloring Agents; Diabetes Mellitus, Experimental; Fibronectins; Glomerulonephritis; Kidney Glomerulus; Laminin; Male; Membrane Glycoproteins; Rats; Rats, Sprague-Dawley

Red blood cells have a negative charge on their surface which prevents Rouloau formation. This charge may reflect the charge on glomerular capillaries. Using alcian blue, we measured the negative charge on red blood cells in patients with IgA nephropathy (IgAN), membranous nephropathy (MN), minimal change nephropathy (MCN) and focal glomerular sclerosis (FGS), for which the diagnosis was determined by renal biopsy. The alcian blue values for the normal control, IgAN, MCN (nephrotic phase), MCN (remission) and MN were 155.3 +/- 12.3, 140.9 +/- 23.9, 101.7 +/- 18.4, 152.9 +/- 11.2 and 140.9 +/- 23.6 ng per 1 x 10(6) red blood cells, respectively. The charge was more significantly decreased in MCN during nephrotic phase than other renal diseases and the normal control. The charge in MCN was within the normal range on remission. When we studied the correlation between the charge on red blood cells and proteinuria during steroid therapy, the charge in MCN was found to increase with reduction of proteinuria, while there was no change in MN and FGS with reduction of proteinuria, i.e. it was usually within normal range. The reduced charge was associated with the nephrotic phase in MCN. The difference in the charge on the red blood cells between MCN and FGS during nephrotic phase seems to suggest a different etiology of proteinuria. Measurement of the charge on red blood cells could be an useful method for differentiating MCN from other renal diseases in the nephrotic phase.

Topics: Adolescent; Adult; Aged; Alcian Blue; Erythrocyte Membrane; Erythrocytes; Female; Glomerulonephritis; Humans; Male; Middle Aged; Nephrosis, Lipoid

The negative charge on the surface of red blood cells may reflect the charge on the glomerular capillaries. Measurements were made of the charge on the red cells of patients with minimal change nephropathy (MCN), membranous nephropathy (IMN), and IgA nephropathy (IgAN) during complete or partial remission. The charge was significantly less in patients with MCN and IMN than in patients with IgAN. Since blood was taken during clinical remission, the differences probably represent the pre-morbid state. Low charge was associated with MCN and IMN, which usually present with heavy proteinuria, and high charge with IgA nephropathy, in which heavy proteinuria is rare. Patients with low red-cell charge are, therefore, susceptible to the development of heavy proteinuria and patients with high charge are protected. The site of deposition or formation of immune complexes may also be determined indirectly by charge.

Topics: Adolescent; Adult; Aged; Alcian Blue; Erythrocyte Membrane; Female; Glomerulonephritis; Glomerulonephritis, IGA; Humans; Immunoglobulin A; Ions; Male; Middle Aged; Nephrosis, Lipoid; Proteinuria; Sex Factors