Albumin Clearance Research Articles

Intravenous endotoxin does not increase tissue extravasation of albumin in rats.

It is unclear whether activation of the inducible nitric oxide synthase (iNOS) increases or decreases the extravasation of plasma. Chloralose anaesthetised male Wistar rats received E. coli lipopolysaccharide (LPS), 3 mg kg-1 i.v., or the corresponding volume of saline, 3 or 5 h before the end of the experiment. Mean arterial pressure (MAP) and heart rate (HR) were recorded. Tissue clearance of radio-labelled albumin, during the last 2 h of each experiment, was determined by a double-isotope method. In separate animals, the serum concentration of nitrite and nitrate was determined, 5 h after LPS or the solvent. LPS initially decreased MAP and lastingly increased HR. In the 3-h LPS animals (n = 8), tissue plasma clearance was lower in the heart and calf muscle and increased only in diaphragm, compared to corresponding control animals (n = 8). In the 5-h LPS rats, clearance was lowered (n = 8) in the entire gastrointestinal tract and in testes, compared to controls (n = 8). The serum nitrite/nitrate concentration was higher in animals given LPS (n = 6) than in controls (n = 6). After LPS, tissue clearance of albumin was not increased in any major tissue, in spite of increased serum levels of NO end products. Apparently, after activation of iNOS, the augmented release of NO is not necessarily associated with increased albumin extravasation.

Doxorubicin pharmacokinetics: the effect of abnormal liver biochemistry tests.

We studied variability in doxorubicin pharmacokinetics in 24 patients with abnormal liver biochemistry tests. Blood samples were collected after the first cycle of single-agent doxorubicin given as an i.v. bolus and plasma levels were measured by high-performance liquid chromatography. The relationship between doxorubicin clearance (dose/AUC) and liver biochemistry tests (AST, bilirubin, albumin, alkaline phosphatase and indocyanine green clearance) was investigated. Patients with a raised bilirubin level had reduced doxorubicin clearance, but there was no clear relationship between the extent of this elevation and the reduction in doxorubicin clearance. Doxorubicin clearance was lower in patients with an isolated increase in AST than in those with normal liver biochemistry, but this difference was not statistically significant. Nevertheless, there was a significant correlation between reduced doxorubicin clearance and both raised serum AST levels and low indocyanine green clearance. These pharmacokinetic data suggest that current dose reductions based solely on the extent to which bilirubin is elevated may not be optimal.

Hypoxia–Reoxygenation Potentiates Zymosan Activated Plasma-Induced Endothelial Injury

The pathophysiology of ischemia–reperfusion injury and the role played by the interaction of plasma proteins, including complement, with reperfused endothelium remains incompletely understood. Venular endothelial changes due to hypoxia followed by reoxygenation (H-R) are vital because venules are the primary site of fluid accumulation and polymorphonuclear leukocyte deposition due to inflammation. This investigation focused on whether H-R potentiates the response to permeability inducing agents found in activated plasma. Activated complement was studied by using zymosan activated plasma (ZAP). Permeability changes were assessed by quantitating rate of clearance of albumin across the monolayers. H-R alone did not change permeability relative to the normoxic condition. ZAP at 2% in normoxic cells increased albumin clearance from 2 ± 0.2 to 9 ± 1.0 μL/h, which increased significantly to 13.5 ± 2.0 μL/h when given to hypoxia–reoxygenation challenged monolayers. The permeability response to ZAP was dose related and not present with heat inactivated ZAP. ZAP at 2% altered the structure of the cytoskeleton of the human umbilical vein endothelial cells (HUVEC). However, addition of monoclonal anti-complement antibodies or addition of soluble complement receptor-1 did not attenuate ZAP-induced HUVEC permeability. Addition of zymosan-activated serum did not alter the permeability and addition of heparin inhibited the ZAP-induced changes in permeability, suggesting that these changes were mediated via thrombin and not complement. The increase in monolayer permeability due to ZAP was prevented by increasing intracellular adenosine-3′,5′-cyclic monophosphate. These findings suggest that HUVEC monolayers challenged with H-R are more susceptible to increases in permeability induced by activated plasma components.

Do obesity and non-insulin dependent diabetes mellitus aggravate exercise-induced microproteinuria?

We evaluated the role of obesity in proteinuria by treadmill exercising of simple obese subjects and non-obese subjects with non-insulin dependent diabetes mellitus in whom the albumin excretion rate at rest was within normal range. Non-obese healthy volunteers were studied as the controls. The fractional renal clearances of four endogenous proteins, albumin, IgG, IgG4, and β2-microglobulin were measured before, during, and after treadmill exercise in 17 simple obese and 15 non-obese diabetic subjects, and in 21 normal subjects. Exercise increased the fractional albumin clearance in all groups. In diabetic subjects, the fractional IgG4 clearance also increased: fractional β2-microglobulin clearance increased in normal controls and in diabetics. In obese subjects, the fractional clearances of albumin, IgG, and IgG4 were similar to those in normal controls, but fractional β2-microglobulin clearance was significantly lower. These results suggest that enhanced microalbuminuria in obese subjects is probably of glomerular origin. In normal subjects and diabetics, exercise-induced microproteinuria is probably of both glomerular and tubular origin. Defect in the charge-selective barrier of the glomerular capillary wall has been implicated in diabetics. Thus some additional factors relevant to obesity must be taken into account in the consideration of the mechanism of microalbuminuria in diabetics with obesity.

Lymph flow from skeletal muscle: Albumin clearance and subsequent appearance in the lymph nodes

The dynamics of local clearance of 99mTc-labelled human serum albumin was compared with the subsequent appearance in the proximal lymph nodes after intramuscular injection into biceps brachialis muscles of five healthy men. Injection site and lymph nodes in the upper arm were imaged by a gamma camera frequently during a 4-hour-period involving both rest and elbow flexion exercises. The percentage of the cleared radioactivity which appeared in the lymph nodes varied among the studied arms but was fairly constant within a given limb, the mean value being 18 %. Qualitatively the local clearance and nodal accumulation appeared in concert, i.e. highest clearance and appearance occurred after muscular exercise. However, there was no significant correlation between total amount of radioactivity cleared and that which appeared in the nodes. We conclude that local clearance of radioactivity provides a better estimate of lymphatic function in healthy subjects than the appearance of radioactivity in the lymph nodes.

Albumin clearance from skeletal muscle depends on the force of contraction

Albumin clearance from skeletal muscle depends on the force of contraction

Tissue and whole-body extracellular, red blood cell and albumin spaces in the rainbow trout as a function of time: a reappraisal of the volume of the secondary circulation

[58Co]EDTA, [51Cr]RBC and [125I]albumin spaces in the whole body and 28 tissue samples were examined at timed intervals over 16 h in rainbow trout Oncorhynchus mykiss. [58Co]EDTA space (which approximates extracellular fluid volume; ECF) in fins, skin, gallbladder and eye are reported for the first time. After a 16 h equilibration, ECF volume was large (376-726 microl g-1 wet tissue mass) in kidney, swimbladder, skin and fins, moderate (219-313 microl g-1 wet tissue mass) in stomach, skull, spleen, liver, intestine, gills, eye and cecum, and small (53-181 microl g-1 wet tissue mass) in red muscle, fat, brain, gallbladder and white muscle. Whole-body ECF was 387+/-10.6 microl g-1 (mean +/- s.e.m.; N=11). [51Cr]RBC space relative to [58Co]EDTA space was large in spleen, liver, intestine and gill, and low in skin, fins, stomach and skull. Whole-body [51Cr]RBC space was 9.9+/-0.6 microl g-1 body mass (N=17). Blood volume calculated from [51Cr]RBC space at 16 h and a dorsal aortic hematocrit of 24.5 % was 40.4 microl g-1 body mass. Whole-body [125I]albumin space at 16 h was 118.0+/-7.4 microl g-1 body mass (N=6), which resulted in an estimated blood volume of 156. 6 microl g-1 body mass, nearly four times that estimated from the [51Cr]RBC space. Tissue hematocrits, calculated from [125I]albumin and [51Cr]RBC spaces, were significantly lower than dorsal aortic hematocrit in all tissues except spleen, kidney and liver. [58Co]EDTA and [51Cr]RBC spaces reached equilibrium in nearly all tissues within 1 h, whereas [125I]albumin continued to accumulate in many tissues up 24 h. The disparity between [125I]albumin distribution kinetics compared with the kinetics of [58Co]EDTA and [51Cr]RBC distribution, as well as the accumulation of [125I]albumin in tissues not known to have a secondary circulation, indicates that [125I]albumin is a poor marker of plasma volume in trout and that previous studies based on [125I]albumin clearance from the plasma have overestimated both the volume and the turnover rate of the secondary system. Revised estimates of secondary circulation volume, based on [58Co]EDTA distribution rate, indicate that it is no more than 10-20 % of the volume of the primary circulation. <P>

Glomerular charge selectivity for horseradish peroxidase and albumin at low and normal ionic strengths.

The classical concept of a negative glomerular charge barrier has recently been questioned, mainly based on the somewhat high clearance for anionic horseradish peroxidase (HRP). The validity of using anionic HRP can be tested by changing the properties of the charge barrier. A rather unequivocal approach is to reduce the ionic composition of the perfusate and hence increase the Debye length. We determined the glomerular clearance for horseradish peroxidase and serum albumin, using isolated rat kidneys perfused at 8 degrees C to reduce the tubular modification of the primary urine. The perfusate contained trace amounts of the neutral 125I-nHRP and the anionic 131I-aHRP and were otherwise identical except for different ionic strengths, 152 mM and 34 mM, respectively. During control, the fractional clearance (theta) was 0.11 +/- 0.015 for nHRP and 0.045 +/- 0.010 for aHRP, with an average clearance ratio (n/a) of 2.8 +/- 0.24. Low ionic strength reduced theta for aHRP to 0.027 +/- 0.006, giving an increased clearance ratio for HRP of 4.2 +/- 0.44. The existence of a negative charge barrier is supported by the experiments. The result obtained during normal perfusion is compatible with a charge density (omega) of 34 mEq L-1, using a model of homogeneously charged membrane. Low ionic strength perfusion reversibly reduced the concentration of fixed charges to 12 mEq L-1, suggesting an almost threefold increase of the glomerular membrane volume. Thus, the glomerular charge barrier should be regarded to have a dynamic gel structure rather than being a rigid membrane.

Glomerular Charge and Size Selectivity Assessed by Changes in Salt Intake in Type 2 Diabetic Patients

To evaluate glomerular charge selectivity in patients with type 2 diabetes, we studied changes in fractional clearance of proteins with different sizes and charges when patients were placed on two diets with different salt contents. Nineteen patients with type 2 diabetes and normoalbuminuria (< 20 micrograms/min, n = 8), microalbuminuria (20-100 micrograms/min, n = 7), or advanced albuminuria (> 100 micrograms/min, n = 4) were placed on a low-salt diet (85 mEq of sodium daily) or a high-salt diet (255 mEq of sodium daily) for 1 week, and then on the other diet, in random order. Fractional clearances of albumin and immunoglobulin G (IgG) were calculated on the last 3 days of each diet. In patients with normoalbuminuria, the high-salt diet increased the fractional clearance of IgG, which is electrically neutral, but the fractional clearance of albumin, which is anionic, was unaltered, suggesting that the pore charge of the glomerular barrier was unaffected. However, in patients with microalbuminuria, the high-salt diet increased the fractional clearances of IgG and albumin equally, indicating some neutralization of the pore charge. Fractional clearance of IgG in these first two groups was similar when salt intake was low, so pore size was the same in these groups. In patients with advanced albuminuria, fractional clearance of IgG was higher than in the other groups, indicating that size selectivity had worsened. In type 2 diabetic patients, charge selectivity is lost before size selectivity as diabetic nephropathy progresses.

Pressure-permeability relationships in basement membrane: effects of static and dynamic pressures.

The glomerular basement membrane (GBM) is an important component of the filtration barrier that is the glomerular capillary wall. Previously GBM permeability has been investigated only under static pressures and often within a supraphysiological range. We used Matrigel as a model of GBM and formed membranes at the base of filtration chamber. We measured membrane permeability under static and dynamic pressures. Matrigel membranes were size and charge selective toward neutrally and negatively charged dextrans. Their permeability (as measured by hydraulic conductivity) was found to decrease from 1.61 +/- 0.06 to 0.75 +/- 0.07 x 10(-6) cm.s-1.cmH2O-1 as static pressure increased from 6 to 78 cmH2O, an effect attributed to membrane compression. In comparison to static pressure, sinusoidal pressure waves with a mean pressure of 50 cmH2O decreased membrane permeability, e.g., fluid flux was reduced by a maximum of 2% to a value of 5.47 +/- 0.38 x 10(-5) cm/s; albumin clearance was reduced by a maximum of 5.2% to a value of 9.63 +/- 1.06 x 10(-6) ml.cm-2.s-1. Such changes were affected by the frequency of pressure wave application and could be attributed to a switching on and off of the membrane compression effect.

Icodextrin with nitroprusside increases ultrafiltration and peritoneal transport during long CAPD dwells

Icodextrin with nitroprusside increases ultrafiltration and peritoneal transport during long CAPD dwells

H2O2causes endothelial barrier dysfunction without disrupting the arginine-nitric oxide pathway

We have previously demonstrated that nitric oxide (.NO) donors attenuate and that inhibition of endogenous nitric oxide synthase (NOS) enhances hydrogen peroxide (H2O2)-mediated porcine pulmonary artery endothelial cell (PAEC) injury. The current study investigates the hypothesis that oxidant-mediated inhibition of NOS contributes to PAEC injury. PAEC barrier function, measured as the transmonolayer clearance of albumin, was significantly impaired by H2O2 (10-100 microM) in the absence of cytotoxicity. Treatment with H2O2 did not alter NOS activity, measured as the conversion of [3H]arginine to [3H]citrulline in PAEC lysates, either immediately after treatment with 0-250 microM H2O2 for 30 min or for up to 120 min after treatment with 100 microM H2O2. H2O2 had little effect on NOS activity in intact PAECs, measured as 1) the formation of [3H]citrulline in [3H]arginine-loaded PAECs, 2) PAEC guanosine 3',5'-cyclic monophosphate content, and 3) PAEC.NO release to the culture media. These results indicate that the arginine-.NO pathway remains intact after exposure to oxidant conditions sufficient to promote functional derangements of vascular endothelial cells.

Role of tyrosine phosphorylation in thrombin-induced endothelial cell contraction and barrier function.

Thrombin-induced endothelial cell (EC) barrier dysfunction is highly dependent upon phosphorylation of serine and threonine residues present on myosin light chains (MLC) catalyzed by a novel EC myosin light chain kinase (MLCK) isoform. In this study, we examined the participation of tyrosine protein phosphorylation in EC contraction, gap formation and barrier dysfunction. We first determined that thrombin significantly increases protein tyrosine kinase activity and protein tyrosine phosphorylation in bovine pulmonary artery EC. Tyrosine kinase inhibitors, genistein and 2,5 DHC, reduced EC tyrosine kinase activities, however, only genistein significantly attenuated thrombin-mediated increases in albumin clearance and reductions in transendothelial electrical resistance. Similarly, genistein but not 2,5 DHC, decreased basal and thrombin-induced Ca2+ increases and MLC phosphorylation in the absence of alterations in Type 1 or 2A serine/threonine phosphatase activities. Immunoprecipitation of the EC MLCK isoform revealed a 214 kD immunoreactive phosphotyrosine protein and genistein pretreatment significantly reduced MLCK activity in MLCK immunoprecipitates. Although thrombin induced the translocation of p60src from the cytosol to the EC cytoskeleton, a detectable increase in the level of MLCK tyrosine phosphorylation was not noted after thrombin challenge. Taken together, our data suggest that genistein-sensitive tyrosine kinase activities are involved in thrombin-mediated EC MLCK activation, MLC phosphorylation, and barrier dysfunction.

Role of 8-epi PGF 2α, 8-isoprostane, in H 2O 2-induced derangements of pulmonary artery endothelial cell barrier function

The non-enzymatic peroxidation product of arachidonic acid, 8-epi-PGF 2α or 8-isoprostane (8-IP) was measured in H 2O 2-exposed cultured pulmonary artery endothelial cell (PAEC) monolayers using a commercially-available enzyme immunoassay kit. H 2O 2 (50 μM for 1–30 min) significantly increased 8-IP production in a time-dependent fashion. Treatment with higher H 2O 2 concentrations (100 or 250 μM) failed to further increase 8-IP generation. Determinations of thiobarbituric acid reactive substances (TBARS) and lipid hydroperoxides (LOOH) were not sufficiently sensitive to detect lipid peroxidation in PAEC exposed to 50 μM H 2O 2 for 15 min. 8-IP (100 pM–500 nM for 2 h) caused PAEC monolayer barrier dysfunction measured as the transmonolayer clearance of albumin without causing significant PAEC cytotoxicity (measured as intracellular lactate dehydrogenase release). This is the first report to provide evidence that 8-IP generated in H 2O 2-exposed PAEC contributes to oxidant-mediated alterations in monolayer barrier function at non-cytotoxic concentrations.

The Influence of Apoptosis on Intestinal Barrier Integrity in Rats

Background: Apoptosis is a critical step responsible for maintaining the cellular balance between proliferation and death and for controlling tumorigenesis. Although an increase in intestinal apoptotic cells has been considered to be associated with the pathogenesis of gastrointestinal injury, little is understood concerning the role of apoptosis in the development of intestinal barrier dysfunction. Methods: Apoptosis induced by intraperitoneal injection of doxorubicin in rats was evaluated by transmission electron microscopy and the TUNEL histochemistry method. Treatment with deoxy-D-glucose (a glycolytic pathway inhibitor) or cycloheximide (a protein synthesis inhibitor) was performed after doxorubicin challenge. Passage of human serum albumin from blood to the intestinal interstitium and the intestinal lumen or from the intestine to the intestinal interstitium and blood was evaluated by means of albumin clearance. Results: A significant increase in gut water content, albumin flux, and bidirectional clearance of albumin accompanied by apoptotic epithelial cell increase was noted in doxorubicin-challenged rats treated with saline. The increase in endothelial and epithelial permeability and the increase of apoptosis could partly be prevented by treatment with deoxy-D-glucose or cycloheximide. Conclusion: Doxorubicin-increased epithelial apoptosis within the intestine occurs simultaneously with increased bidirectional permeability of the intestinal barrier, probably associated with both glycolytic and protein synthesis pathways. Apoptosis may thus play a role in the pathogenesis of intestinal barrier dysfunction.

Fractional Clearance of Albumin is Influenced by its Degradation during Renal Passage

1. The fractional clearance of intact albumin as determined by fractionation of urine by gel chromatography gave a value of 3.9 +/- 1.6 x 10(-4) for the isolated perfused kidney and 2.1 +/- 0.6 x 10(-4) in vivo using ALZET osmotic pumps. 2. Albumin fractional clearance as measured by detection of the tritium label on the albumin molecule by radioactivity analysis gave a value of 7.5 +/- 3.9 x 10(-3) for the isolated perfused kidney and 2.3 +/- 0.9 x 10(-3) in vivo. 3. The major differences between assays that detect intact albumin compared with non-specific assays in the estimates of the fractional clearance of albumin can be explained by the degradation of approx. 90% of albumin to small peptides during its renal passage. This has been demonstrated by size exclusion chromatography of urine samples from experiments where (i) exogenous tritium-labelled albumin was used in isolated perfused kidneys, (ii) exogenous tritium-labelled albumin was administered intravenously and (iii) analysis was made with metabolically labelled endogenous albumin in vivo.

Impaired autoregulation of GFR in hypertensive non-insulin dependent diabetic patients

We investigated the effect of acute lowering of blood pressure (BP) upon glomerular filtration rate (GFR) in hypertensive non-insulin-dependent diabetes mellitus (NIDDM) patients, 14 with diabetic nephropathy and 12 with normoalbuminuria. The study was performed twice with the subjects receiving an intravenous injection of either clonidine (150 to 225 micrograms) or saline (0.154 mmol/liter). We assessed GFR, albuminuria, and BP. The two groups were well matched with respect to demographic data, baseline GFR and BP. Clonidine induced similar reductions in mean arterial blood pressure 19 (SE +/- 4) and 21 (SE +/- 3) mm Hg in patients with and without nephropathy, respectively. In the nephropathy group GFR diminished in average from 90 (SE +/- 6) to 81 (SE +/- 7) ml/min/1.73 m2 (P = 0.006), fractional clearance of albumin (x 10(-6)) declined from a geometric mean of 219 (antilog SE /divided by 1.3) to 186 (antilog SE /divided by 1.3) (P = 0.04), and four patients had a complete pressure-passive vasculature, defined as delta GFR% = delta MABP%. A significant correlation between relative reductions in MABP and GFR (r = 0.78, P < 0.001) was demonstrated in albuminuric patients. None of the normoalbuminuric patients had a complete pressure-passive vasculature and there were no significant differences in GFR between the two examinations, but five had abnormal autoregulation of GFR. Mean difference between changes in GFR (95% confidence interval) between the nephropathic and normoalbuminuric group was 5.5 (divided by 2.7 to 13.7) ml/min/1.73 m2 (P = 0.18). Our study suggests that hypertensive NIDDM patients, particularly patients with nephropathy, frequently suffer from impaired or abolished autoregulation of GFR.

Protein, not adenosine or adenine nucleotides, mediates platelet decrease in endothelial permeability.

Platelets and platelet-conditioned medium (PCM) decrease endothelial protein permeability in vitro. Adenosine and a > 100-kDa protein have previously been implicated as the soluble factors released from platelets that decrease endothelial permeability. The objective of this study was to further investigate the role of adenosine in this platelet response. Measurements of adenosine and its precursor adenine nucleotides by high-performance liquid chromatography were correlated with the assessment of permeability by 125I-labeled albumin clearance and electrical resistance across endothelial cell monolayers derived from the bovine pulmonary artery. PCM contained micromolar concentrations of AMP, ADP, and ATP, but adenosine was below detectable levels (< or = 0.1 microM). Adenosine deaminase, an enzyme that converts adenosine to inactive inosine, or an adenosine-receptor antagonist did not block the platelet- or PCM-mediated decrease in endothelial permeability. A < 3-kDa fraction of PCM that contained micromolar concentrations of AMP and ADP did not affect endothelial permeability, whereas a > 3-kDa fraction that contained much reduced levels of AMP and ADP significantly decreased permeability. This activity of PCM was sensitive to insoluble trypsin. This study rules out adenosine and adenine nucleotides as primary factors in the platelet-induced decrease in endothelial permeability and suggests that the active factor is a protein.

Site-specific thrombin receptor antibodies inhibit Ca2+ signaling and increased endothelial permeability.

Thrombin receptor is activated by thrombin-mediated cleavage of the receptor's NH2 terminus between Arg-41 and Ser-42, generating a new NH2 terminus that functions as a "tethered ligand" by binding to sites on the receptor. We prepared antibodies (Abs) directed against specific receptor domains to study the tethered ligand-receptor interactions required for signaling the increase in endothelial permeability to albumin. We used polyclonal Abs directed against the peptide sequences corresponding to the extracellular NH2 terminus [residues 70-99 (AbDD) and 1-160 (AbEE)] and extracellular loops 1 and 2 [residues 161-178 (AbL1) and 244-265 (AbL2)] of the seven-transmembrane thrombin receptor. Receptor activation was determined by measuring changes in cytosolic Ca2+ concentration ([Ca2+]i) in human dermal microvascular endothelial cells (HMEC) loaded with Ca(2+)-sensitive fura 2-acetoxymethyl ester dye. The transendothelial 125I-labeled albumin clearance rate (a measure of endothelial permeability) was determined across the confluent HMEC monolayers. AbEE (300 micrograms/ml), directed against the entire extracellular NH2-terminal extension, inhibited the thrombin-induced increases in [Ca2+]i and the endothelial 125I-albumin clearance rate (> 90% reduction in both responses). AbDD (300 micrograms/ml), directed against a sequence within the NH2-terminal extension, inhibited 70% of the thrombin-induced increase in [Ca2+]i and 60% of the increased 125I-albumin clearance rate. AbL2 (300 micrograms/ml) inhibited these responses by 70 and 80%, respectively. However, AbL1 (300 micrograms/ml) had no effect on either response. We conclude that NH2-terminal extension and loop 2 are critical sites for thrombin receptor activation in endothelial cells and thus lead to increased [Ca2+]i and transendothelial permeability to albumin.

Effects of supplementation with vitamin C or E on albuminuria, glomerular TGF-beta, and glomerular size in diabetes.

Oxidant stress and a reduction in antioxidant status, including reduced plasma and tissue ascorbic acid content, occur in diabetic patients and experimental models of diabetes. In this study, the effects of treatment of streptozotocin diabetic rats for 2 mo with vitamin C (10 g/kg body wt/d) or dietary vitamin E (200 mg/kg body wt/d) in the drinking water on urinary albumin excretion, glomerular transforming growth factor (TGF)-beta content, and glomerular size were examined. Treatment of diabetic rats with vitamin C or E had no effect on blood glucose levels compared with that in untreated diabetics (453 +/- 28 g/dl +/- SEM). Body weight, BP, and creatinine clearance rates were not significantly different among the study groups. Kidney weight was significantly higher in all of the diabetic groups compared with age-matched control rats. Treatment with vitamin C, but not vitamin E, significantly reduced kidney weight compared with that in untreated diabetic rats. Immunohistochemical staining for TGF-beta was 2.5-fold higher in glomeruli of cortical sections from untreated diabetic rats versus control rats. Treatment with vitamin C or E prevented the increase in glomerular TGF-beta immunoreactivity. Glomerular volume was also significantly increased (twofold) in kidneys of untreated diabetic rats compared with control rats, as assessed by light microscopy. Treatment with vitamin C prevented and treatment with vitamin E reduced the increase in glomerular volume. Treatment with vitamin C also prevented the sevenfold increase in albumin clearance otherwise seen in untreated diabetic rats. By contrast, treatment with vitamin E had no effect on albumin clearance despite reductions in glomerular size and TGF-beta. Renal cortical vitamin E and plasma, but not renal cortical vitamin C, were reduced in diabetic rats versus control rats. Supplementation of diabetic rats with vitamin C markedly increased plasma and renal cortical vitamin C content to values greater than those in control rats. Supplementation with vitamin E increased renal cortical vitamin E content by 50% compared with values in control rats and also increased plasma and renal cortical vitamin C. These results support the potential utility of antioxidant treatment for the prevention of renal injury in diabetes.