thromboplastin and Proteinuria

Glomerular inflammation is a putative aggravation factor for type 2 diabetic nephropathy and urinary thrombin is a novel marker of glomerular inflammation. To clarify the relationship between glomerular inflammation and progression of the nephropathy, we measured urinary thrombin in 118 patients with type 2 diabetic nephropathy at different stages. To investigate the implications of urinary thrombin in the nephropathy, we compared urinary thrombin with expression of tissue factor, the trigger of blood coagulation activation, in glomeruli and with markers of renal injury (estimated glomerular filtration rate (eGFR) and proteinuria). Urinary thrombin was found in 4.9% (3/61), 0.0% (0/12), 29.6% (8/27) and 50.0% (9/18) of patient groups at stages 1, 2, 3 and 4, respectively. Thus, urinary thrombin was negligible in the patients at early stages (stages 1 and 2), but was present predominantly in the patients at advanced stages (stages 3 and 4). Tissue factor was expressed in accumulated macrophages in glomeruli, which indicates that thrombin may be generated in inflamed glomeruli presumably via inflammation-induced activation of the exudated coagulation factors into glomerular tissues and then be excreted in urine. Urinary thrombin was significantly associated with both decreased eGFR and increased proteinuria in type 2 diabetic nephropathy. Therefore, increased urinary thrombin in patients with advanced stages of type 2 diabetic nephropathy suggests that glomerular inflammation may injure the tissues, thereby impairing renal function. Monitoring an effect of anti-diabetic treatments on glomerular inflammation in the patients with type 2 diabetic nephropathy may be a possible application of urinary thrombin.

Topics: Antithrombin III; Biomarkers; C-Reactive Protein; Diabetes Mellitus, Type 2; Female; Glomerular Filtration Rate; Humans; Inflammation; Kidney Glomerulus; Male; Middle Aged; Peptide Hydrolases; Proteinuria; Thrombin; Thromboplastin

Tissue factor (TF) is a transmembrane protein that is essential for coagulation. TF is expressed on podocytes and its cytoplasmic domain has cell signalling functions in epithelial cells.. Mice lacking the cytoplasmic domain of TF (TF(CT-/-) mice) were used to study its role in physiological albuminuria and pathological proteinuria following induction of glomerulonephritis (GN).. Absence of the cytoplasmic domain of TF was associated with increased albuminuria, podocyte effacement, reduced podocyte numbers and increased spontaneous glomerular tumour necrosis factor α(TNFα) production under physiological conditions. In mice developing GN, absence of the cytoplasmic domain of TF resulted in increased proteinuria and enhanced renal TNFα production without altering other parameters of renal inflammation and injury. Studies in TF(CT-/-) chimeric mice (created by bone marrow transplantation) showed increased proteinuria and renal TNFα mRNA in GN was associated with absence of the cytoplasmic domain of TF in the kidney and was independent of the leucocyte phenotype.. These studies demonstrate that the cytoplasmic domain of TF contributes to renal albumin retention and its renal expression protects against proteinuria in leucocyte-mediated renal inflammation. Increased glomerular production of TNFα in the absence of cytoplasmic domain of TF may contribute to podocyte injury resulting in albuminuria and proteinuria.

Topics: Albuminuria; Animals; Cytoplasm; Glomerulonephritis; Intracellular Signaling Peptides and Proteins; Kidney Glomerulus; Membrane Proteins; Mice; Mice, Inbred C57BL; Mice, Knockout; Podocytes; Protein Structure, Tertiary; Proteinuria; Thromboplastin; Tumor Necrosis Factor-alpha

Nephrotic syndrome (NS) is associated with numerous blood coagulation abnormalities and a marked predisposition to thromboembolism. Fibrin formation within the glomeruli occurs in various forms of human and experimental glomerulonephritis and may play an important role in progressive glomerular injury. The aim of this study was to measure the plasma concentrations of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) and intravascular thrombin generation markers and to analyze their relationships in patients with primary glomerulonephritis.. The study population comprised 57 patients (mean age 35.2 years; range 18-63 years) with primary glomerulonephritis: 36 with NS (NS group) and 21 without (non-NS group). The control group consisted of 24 sex- and age-matched healthy volunteers. TF and TFPI antigen, prothrombin fragment F 1+2 (PF 1+2) and thrombin-antithrombin III complex (TAT) concentrations in plasma were estimated using commercially available kits.. Serum TF and TFPI concentrations in both the NS and non-NS groups were higher than those observed in the control group. Moreover, there were significant differences in TF and TFPI concentrations between the NS and non-NS groups. TF:TFPI ratios in both the examined groups were constant and significantly higher than those in the control group. Positive correlations between TF and both PF 1+2 and TAT concentrations in the total cohort of patients were shown. Furthermore, a positive correlation between TF and TFPI concentrations was observed.. Our data support the hypothesis concerning activation of coagulation pathways in patients with primary glomerulonephritis. An inadequate TFPI concentration as a result of an elevated TF:TFPI ratio characterizes not only patients with clinical manifestations of NS but also patients with mild proteinuria.

Topics: Adolescent; Adult; Antithrombin III; Blood Coagulation; Case-Control Studies; Disease Progression; Female; Glomerulonephritis; Humans; Lipoproteins; Male; Middle Aged; Nephrotic Syndrome; Peptide Fragments; Peptide Hydrolases; Protein Precursors; Proteinuria; Prothrombin; Thromboplastin

Topics: Adult; Aged; Angiotensin-Converting Enzyme Inhibitors; Biomarkers; Case-Control Studies; Endothelium, Vascular; Female; Humans; Kidney Failure, Chronic; Lipoproteins; Male; Middle Aged; Proteinuria; Thrombomodulin; Thromboplastin; von Willebrand Factor

To investigate factors that influence urinary tissue factor (uTF) measurements: glomerular permeability and filtration, tubular function, haematuria, and urine bacterial growth.. uTF, protein creatinine index, glomerular filtration rate, retinol binding protein, N-acetyl-beta-D-glucosaminidase (NAG) and urinary haemoglobin (uHb) were measured in patients with hypertension, diabetes mellitus and nephrotic syndrome (n = 342), tubulo-interstitial disease (n = 50), and haematuria of uncertain cause (n = 50); measurements were also made in urine samples from healthy subjects for "simulated" haematuria (n = 6) and bacterial growth (n = 4) studies.. There was a weak correlation of uTF with glomerular permeability and filtration (protein creatinine index and glomerular filtration rate) and with markers of tubular function (retinol binding protein and NAG). uTF concentrations were not affected by the presence of blood or bacteria in the urine sample.. uTF concentrations are relatively stable. This is an important finding if the assay is to be used in clinical practice.

Topics: Acetylglucosaminidase; Bacteriuria; Biomarkers; Glomerular Filtration Rate; Hematuria; Humans; Kidney; Kidney Tubules; Proteinuria; Retinol-Binding Proteins; Thromboplastin

Increased glomerular tissue factor (TF) expression is associated with glomerular fibrin deposition and renal failure in human and experimental crescentic glomerulonephritis (GN). However, the in vivo functional contribution of TF to the development of glomerular fibrin deposition, crescent formation, and renal failure in GN has not been established. The contribution of TF to fibrin deposition and renal injury was studied in a rabbit model of crescentic GN in which glomerular macrophage infiltration, augmented TF expression, and fibrin deposition are prominent. Administration of anti-TF antibody inhibited glomerular TF activity in nephritic glomeruli by 96%, without affecting macrophage accumulation or systemic indices of coagulation. Anti-TF antibody significantly reduced glomerular fibrin deposition (fibrin scores, 0.43 +/- 0.10 (treated) and 1.40 +/- 0.19 (control); P < 0.0005), crescent formation (0.33 +/- 0.05 (treated) and 1.0 +/- 0.06 (control); P < 0.0005), and development of renal failure (serum creatinine, 168 +/- 22 mumol/l (treated) and 267 +/- 35 mumol/l (control); P < 0.04). This was associated with significant reduction in proteinuria (1189 +/- 277 mg/24 hours (treated) and 2060 +/- 336 mg/24 hours (control); P < 0.03) and expression of MHC class II antigen in glomeruli (1.25 +/- 0.41 (treated) and 2.83 +/- 0.53 (control); P < 0.03) and in tubules and interstitial areas. These data demonstrate that TF is the major in vivo initiator of fibrin deposition in crescentic GN. The reduction in proteinuria and glomerular major histocompatibility class II antigen expression by TF inhibition suggests that TF may also activate other mediators that contribute to glomerular injury.

Topics: Animals; Antibodies; Blood Coagulation; Fibrin; Glomerulonephritis; Histocompatibility Antigens Class II; Immunoconjugates; Kidney Glomerulus; Male; Proteinuria; Rabbits; Renal Insufficiency; Thromboplastin

To investigate the significance of urinary tissue factor (uTF) concentrations in patients with glomerulonephritis.. Urine samples were collected from normal subjects (n = 57), patients with uncomplicated renal stones (n = 30), and patients with glomerulonephritis (n = 150). Samples were then centrifuged and the pellets solubilised in n-octyl-beta-glucopyranoside. uTF concentrations were determined using a one stage kinetic chromogenic assay.. The uTF concentration was higher in patients with glomerulonephritis than in normal controls (p < 0.01) or in patients with renal stones (p < 0.05). uTF activity correlated with the protein creatinine index (PCI, r = 0.41, p < 0.001) and seven patients with glomerulonephritis and a PCI < or = 0.1 g/mmol had raised uTF. Glomerulonephritis patients were subdivided into two groups depending on the PCI: < 0.2 g/mmol creatinine (mild to moderate proteinuria, group I) and > or = 0.2 g/mmol creatinine (heavy proteinuria, group II). In group I, uTF concentrations were higher in patients with either immune complex (IC) glomerulonephritis (p < 0.01) or non-IC (p < 0.05) glomerulonephritis than in normal controls. In group II, the IC glomerulonephritis group had higher uTF concentrations than normal controls (p < 0.001) or patients with renal stones (p < 0.01); and non-IC glomerulonephritis patients had higher uTF than normal controls (p < 0.01). When the glomerulonephritis groups were divided into broad WHO subtypes, the significance level varied with the type of glomerulonephritis.. uTF is increased in patients with glomerulonephritis, and its concentration may reflect the aetiopathogenesis of glomerulonephritis.

Topics: Biomarkers; Creatinine; Glomerulonephritis; Humans; Kidney Calculi; Proteinuria; Thromboplastin

We tested the hypothesis that enhanced intravascular coagulation in pregnancy could produce clinical symptoms similar to those of preeclampsia, such as hypertension, proteinuria, and edema. Having confirmed this, we then examined whether the pathological changes caused by intravascular coagulation could be suppressed by administration of antithrombin III (AT III), an endogenous inhibitor active to thrombin and factor X a. Intravascular coagulation was induced in Wistar rats on day 16-20 of pregnancy by 1-h arterial infusion of tissue thromboplastin (TP) through the left ventricle of the heart. One hour after the end of the infusion period, organ blood flows were measured by the radioactive ((57)Co-labeled) microsphere method, and fibrin deposition in organs was measured by radiolabeling with [(125)I] fibrinogen injected before TP infusion. Infusion of TP produced fibrin deposition in the kidney, lung, and liver, but not in the myometrium and placenta, as well as an 80% decrease in renal blood flow (RBF), with oliguria and proteinuria. TP also caused an increase in blood pressure (BP) accompanied by an increase in plasma renin activity (PRA), both of which were suppressed by bilateral nephrectomy before TP infusion. The prophylactic administration of AT III concentrates (60 or 300 U/kg intravenously (i.v.), followed by infusion of 30 or 150 U/kg/2 h, respectively) prevented all pathological changes in a dose-dependent manner. AT III increased placental blood flow regardless of the state of coagulation. These findings suggest that intravascular coagulation plays a significant part in the pathophysiology of preeclampsia and that AT III concentrates may have therapeutic potential in the treatment of this condition.

Topics: Animals; Antithrombin III; Blood Coagulation; Female; Fibrin; Hypertension; Kidney; Pre-Eclampsia; Pregnancy; Pregnancy Complications, Cardiovascular; Proteinuria; Rats; Rats, Wistar; Regional Blood Flow; Renin; Serum Albumin; Thromboplastin

Correlations between glomerular expression of tissue factor (TF) activity and antigen and cellular localization of TF mRNA was studied in crescentic glomerulonephritis (GN) in rabbits. Glomerular TF activity increased 8.7-fold 24 hours after initiation of GN (234 +/- 49 mU/10(3) glomeruli; normal, 27 +/- 10 mU/10(3) glomeruli; P = 0.003) in association with a 2.1-fold increase in TF antigen (154 +/- 34 ng/10(3) glomeruli; normal, 72 +/- 10 ng/10(3) glomeruli; P = 0.055), early macrophage infiltration, and no significant increase in TF mRNA. At the peak glomerular macrophage infiltration (day 4), TF activity remained augmented (230 +/- 63 mU/10(3) glomeruli) and TF mRNA, colocalized within macrophages, was significantly increased compared with normal (267 +/- 42%; P = 0.001). TF antigen was not increased in glomeruli (114 +/- 17 ng/10(3) glomeruli), although significant urinary excretion of TF antigen was detectable (478 +/- 121 ng/24 hours; normal, < 1 ng/24 hours; P = 0.032). At this time, the M(r) of glomerular TF (49 to 61 kd) was increased compared with TF in normal glomeruli (49 to 58 kd) as a result of increased glycosylation. At day 7, TF activity and antigen within glomeruli had decreased, although urinary excretion of TF antigen and glomerular TF mRNA remained elevated. These studies suggest that early up-regulation of TF activity is largely a result of functional up-regulation of constitutive TF in intrinsic glomerular cells. In more advanced disease, infiltrating macrophages are the major site of TF synthesis. The increased M(r) of glomerular TF, as a result of synthesis of more highly glycosylated protein by macrophages and the shedding of TF into the urine, suggests that substantial turnover of glomerular TF occurs at this stage.

Topics: Animals; Antibodies, Monoclonal; Glomerulonephritis; In Situ Hybridization; Kidney Glomerulus; Male; Proteinuria; Rabbits; RNA, Messenger; Thromboplastin

We studied the relationships between albuminuria, tissue factor-induced coagulation, and endothelial cell dysfunction in 67 patients with non-insulin-dependent diabetes mellitus (NIDDM) who were divided into three groups on the basis of their urinary albumin excretion rate (AER). To assess the early phase of tissue factor-induced coagulation, activated factor VII (FVIIa) levels in plasma were measured by a direct fluorogenic assay. As markers of endothelial cell dysfunction, levels of von Willebrand factor (vWF), tissue-type plasminogen activator-plasminogen activator inhibitor-1 (TPA-PAI-1) complex, PAI-1, and tissue factor pathway inhibitor (TFPI) were measured. FVIIa levels were increased in normoalbuminuric NIDDM patients (AER < 15 micrograms/min) when compared with normal control subjects. This FVIIa increase was accompanied by an increase in thrombin-antithrombin III complex (TAT) levels, indicating increased activation of coagulation even in normoalbuminuric patients. In NIDDM patients with microalbuminuria (AER = 15-200 micrograms/min), the FVIIa level, the FVIIa-FVII antigen (Ag) ratio (an indicator of activation of FVII zymogen to FVIIa), and the TAT level were further increased. This group also had higher levels of endothelial cell-derived factors (vWF, TPA-PAI-1 complex, and PAI-1) than the control group. The levels of endothelial cell-derived factors (including TFPI) were highest in the NIDDM patients with overt albuminuria (AER > 200 micrograms/min). In all 67 diabetic patients, AER showed a strong positive correlation with FVIIa (r = .574, P < .0001) and a weakly but still significant correlation with FVIIa-FVII:Ag (r = .365, P = .01), vWF (r = .315, P < .01), and TAT (r = .323, P < .01).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Aged; Blood Coagulation; Blood Pressure; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Electrocardiography; Endothelium, Vascular; Enzyme Activation; Factor VII; Female; Heart Diseases; Humans; Male; Middle Aged; Proteinuria; Thromboplastin; von Willebrand Factor

The mechanism involved in glomerular fibrin deposition was investigated during mercuric chloride (HgCl2)-induced autoimmune glomerulonephritis in the Brown Norway rat. To ascertain whether the local hemostatic system was activated secondarily to the immunological conflict, the ability of glomerular lysates to induce coagulation in vitro was assessed in treated and control rats. Glomerular procoagulant activity (PCA) of HgCl2-injected rats was measured on day 12 (latent phase of the disease), day 20 (acme), and days 32 and 42 (recovery phase) after the first mercury injection. PCA rose 3-fold (p less than 0.02) at day 20 and then almost returned to control values. Proteinuria, PCA, and the incidence of glomerular fibrin deposits peaked concomitantly at day 20. Glomerular PCA was characterized as thromboplastin. The number of Ia positive cells detected by monoclonal OX-6 antibody was not different from the control number at any phase of the disease; the number of macrophages per glomerular section detected by electron microscopy at day 20 in HgCl2-injected rats was 1.80 +/- 0.60, versus 0.30 +/- 0.11 in the controls. No correlation was found between glomerular PCA and either the number of monocytes/macrophages or of Ia-positive cells present in the glomeruli. Since glomerular PCA was maximal at the onset of fibrin formation in the glomeruli and then decreased toward its basal level, and since the fibrin disappeared, it is concluded that increased production of thromboplastin by glomeruli, with activation of the extrinsic coagulation pathway, may contribute to intraglomerular fibrin deposition in HgCl2-induced glomerulonephritis.

Topics: Animals; Autoimmune Diseases; Female; Fibrin; Fibrin Fibrinogen Degradation Products; Glomerulonephritis; Histocompatibility Antigens Class II; Kidney Glomerulus; Macrophages; Male; Mercuric Chloride; Proteinuria; Rats; Rats, Inbred BN; Thromboplastin

Four patients had clinical manifestations of the hemolytic-uremic syndrome. No evidence of active intravascular coagulation was found during the acute phase of the illness, using a sensitive assay to measure soluble circulating fibrin in the plasma of these patients, three of whom developed the clinical syndrome while hospitalized for gastro-enteritis. These findings, coupled with the findings of others, suggest that either the episode of intravascular coagulation precedes the development of the clinical manifestations, or that platelet thrombosis is occurring in the absence of activation of plasma clotting factors. In any case, heparin anticoagulant therapy does not seem indicated.

Topics: Anuria; Blood Cell Count; Blood Coagulation Tests; Blood Platelets; Blood Pressure; Blood Urea Nitrogen; Carbon Radioisotopes; Child; Child, Preschool; Creatinine; Disseminated Intravascular Coagulation; Factor V; Factor VIII; Female; Fibrin; Fibrinogen; Hematuria; Hemoglobins; Hemolytic-Uremic Syndrome; Heparin; Humans; Infant; Male; Proteinuria; Prothrombin Time; Thromboplastin; Thrombosis

Topics: Adolescent; Anticoagulants; Blood Coagulation Disorders; Blood Coagulation Tests; Factor IX; Furosemide; Glucocorticoids; Hemophilia B; Humans; Immune Sera; Immunization; Immunoelectrophoresis; Male; Nephrotic Syndrome; Prednisone; Proteinuria; Prothrombin Time; Thromboplastin

Topics: Animals; Blood Platelets; Edema; Female; Humans; Hypertension, Renal; Immune Sera; Kidney Glomerulus; Kidney Tubules; Microscopy, Electron; Placenta; Pre-Eclampsia; Pregnancy; Proteinuria; Rabbits; Regional Blood Flow; Thromboplastin