thromboplastin and Peritonitis

TFPI-tissue factor pathway inhibitor appears to play a primary role in regulating TF-induced coagulation. During CAPD procoagulant and anticoagulant activities of the mesothelium are balanced under normal conditions. The aim of the work was to assess TF and TFPI concentrations during peritonitis in CAPD patients. The study was performed in 9 CAPD subjects with peritonitis and 14 clinically stable CAPD patients. TF, total, free and truncated TFPI, thrombomodulin concentrations were measured in plasma and dialysate; C-reactive protein and tumor necrosis factor-TNF alpha were assayed in serum. In 8 patients with S. aureus peritonitis TF and TFPI were not found in dialysate, but were detectable in a case with E. coli peritonitis. C-reactive protein and TNF alpha significantly elevated at the beginning of peritonitis, fell sharply after the therapy. Further studies are needed to establish whether the kind of bacterial peritonitis (Gram-positive or negative) may affect TF and TFPI in plasma and dialysate in CAPD patients.

Topics: Adult; Aged; Anticoagulants; Bacterial Infections; C-Reactive Protein; Dialysis Solutions; Humans; Lipoproteins; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; Staphylococcal Infections; Thrombomodulin; Thromboplastin; Tumor Necrosis Factor-alpha

Topics: Animals; Anti-Bacterial Agents; Anti-Infective Agents; Blood Coagulation; Coagulase; Female; Fibrin; Fibrinogen; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Peritonitis; Prothrombin; Staphylococcal Infections; Staphylococcus aureus; Thromboplastin

Inhibition of blood coagulation appears to be an important therapeutic strategy to improve the outcome in sepsis. However, the beneficial effect of anticoagulant treatment in sepsis is solely based on experimental data using inhibitors of the extrinsic coagulant pathway. The role of the intrinsic pathway of coagulation in the pathogenesis of sepsis has not been explored yet.. In the current study, we contribute to determine the role of factor (F)VIII, the key player of the intrinsic coagulant pathway, on host defense against peritonitis.. Hemizygous FVIII-deficient mice and their wild-type littermates were challenged with 1 x 10(4) bacteria in a septic peritonitis model.. The intraperitoneal injection of Escherichia coli led to growth and dissemination of bacteria and provoked an inflammatory response as evident from elevated cytokine levels, increased cell influx into tissues, liver necrosis, and endothelialitis resulting in mortality. The FVIII-deficient genotype slightly reduced bacterial outgrowth but had no effect on markers of inflammation and/or survival. In addition, FVIII-deficient mice showed profound activation of coagulation, thereby improving the hemophilic phenotype of FVIII-deficient mice.. FVIII deficiency slightly modifies host defense in septic peritonitis in mice, but does not influence the final outcome of peritonitis. Therefore, we question the importance of the intrinsic coagulant pathway during sepsis.

Topics: Animals; Blood Coagulation; Colony Count, Microbial; Escherichia coli; Factor VIII; Hemophilia A; Immunity; Inflammation; Lipoproteins; Mice; Peritonitis; RNA, Messenger; Sepsis; Survival Rate; Thromboplastin

Anticoagulants have gained increasing attention for the treatment of sepsis. Inhibition of the tissue factor (TF)/factor (F) VIIa pathway has been shown to attenuate the activation of coagulation and to prevent death in a primate model of sepsis caused by gram-negative bacteria.. To determine the role of the TF/FVIIa complex in the host response to peritonitis, mice received an intraperitoneal injection of live Escherichia coli with or without concurrent treatment with recombinant nematode anticoagulant protein c2 (rNAPc2), a selective inhibitor of the TF/FVIIa pathway.. Peritonitis was associated with an increase in the expression of TF at the tissue level and activation of coagulation, as reflected by elevated levels of thrombin-antithrombin complexes and by increased fibrin(ogen) deposition in the liver and lungs. rNAPc2 strongly attenuated this procoagulant response but did not influence the inflammatory response (histopathology, leukocyte recruitment to the peritoneal cavity, and cytokine and chemokine levels). Moreover, rNAPc2 did not alter bacterial outgrowth locally or dissemination of the infection, and survival was not different between rNAPc2-treated mice and control mice.. These data suggest that TF/FVIIa activity contributes to the activation of coagulation during E. coli peritonitis but does not play a role in the inflammatory response or antibacterial host defense.

Topics: Animals; Blood Coagulation; Disease Models, Animal; Escherichia coli; Escherichia coli Infections; Factor VIIa; Helminth Proteins; Inflammation; Male; Mice; Mice, Inbred C57BL; Peritonitis; Thromboplastin

Patients treated with peritoneal dialysis frequently suffer from recurrent peritonitis episodes. During peritonitis, inflammatory mediators are released and a serofibrinous exudate is formed in the peritoneal cavity, which promotes fibrosis and abdominal adhesion development. Human peritoneal mesothelial cells (HMC) play a critical role in maintaining the intraperitoneal balance between fibrinolysis and coagulation by expressing the fibrinolytic enzyme tissue-type plasminogen activator (t-PA) and its specific inhibitor, plasminogen activator inhibitor-1 (PAI-1) as well as the procoagulant protein, tissue factor.. Cultured HMC were used to examine the effect of a 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, simvastatin, on the expression of t-PA, PAI-1 and tissue factor after activation of the cells with tumor necrosis factor-alpha (TNF-alpha). Antigen concentrations in the cell supernatants were measured by enzyme-linked immunosorbent assay (ELISA). Northern blot analysis was conducted for mRNA expression. Luciferase reporter gene assays and Western blot analysis in human fibrosarcoma HT1080 cells and HMC were performed to analyze the effect of simvastatin on the transcription factors nuclear factor kappa B (NF-kappa B) and activator protein-1 (AP-1), which regulate tissue factor gene expression.. Incubation of HMC with TNF-alpha resulted in significantly decreased t-PA and increased PAI-1 synthesis. In the presence of simvastatin t-PA synthesis in control and TNF-alpha-treated cells dose-dependently increased, reaching 5.8-fold and 7.7-fold higher t-PA levels, respectively, at 5 micromol/L simvastatin after 48 hours. Simvastatin dose-dependently suppressed PAI-1 production in both control and TNF-alpha-treated cells. At 5 micromol/L, simvastatin lowered PAI-1 synthesis 3.4-fold and 4.0-fold, respectively, thereby also completely suppressing the TNF-alpha effect itself. Similarly, simvastatin down-regulated the expression of tissue factor and also completely opposed the TNF-alpha-induced tissue factor expression. The effects of simvastatin on t-PA, PAI-1 and tissue factor expression were prevented by mevalonate and geranylgeraniol (GG), suggesting the involvement of geranylgeranyl-modified intermediates in simvastatin's mode of action. Also, simvastatin reduced NF-kappa B- and AP-1-dependent reporter gene activity in TNF-alpha-treated HT-1080 fibrosarcoma cells and reduced the nuclear levels of p50-NF-kappa B, p65-NF-kappa B, and the AP-1 components c-fos and c-jun in HMC.. The HMG-CoA reductase inhibitor simvastatin is an effective stimulator of the mesothelial fibrinolytic capacity and suppresses the procoagulant activity both under normal and inflammatory conditions. Our findings provide a molecular explanation for the anti-inflammatory properties of statins in HMC and a rationale for the use of these drugs to protect peritoneal dialysis patients from peritoneal fibrosis and adhesion development during bacterial peritonitis.

Topics: Antineoplastic Agents; Cells, Cultured; Diterpenes; Epithelial Cells; Epithelium; Fibrinolysis; Gene Expression; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Mevalonic Acid; Omentum; Peritoneal Dialysis; Peritoneum; Peritonitis; Plasminogen Activator Inhibitor 1; Simvastatin; Thromboplastin; Tumor Necrosis Factor-alpha

To determine whether treatment with recombinant human tissue factor pathway inhibitor (TFPI), an inhibitor of the extrinsic coagulation pathway, can improve survival in a clinically relevant model of gram-negative sepsis, rabbits were given an intraperitoneal inoculation of a suspension containing hemoglobin (40 microg/mL), porcine mucin (150 microg/mL), and viable Escherichia coli O18:K1 (1.0 +/- 0.5 x 10(5) cfu/kg). Treatment with gentamicin (5 mg/kg every 12 h for five doses) was instituted 4 h after induction of peritonitis. At the same time point, rabbits were randomized to receive a 24-h infusion of vehicle or one of three different doses of TFPI. Treatment groups, 7-day survival rates, and significance versus control were as follows: control, 1 of 20; TFPI(LOW DOSE) (0.1 mg/kg, then 1 microg/kg/min), 3 of 12 (P = .14); TFPI(MID DOSE), (0.5 mg/kg, then 5 microg/kg/min), 7 of 12 (P = .002); TFPI(HIGH DOSE) (10 mg/kg, then 10 microg/kg/min), 4 of 13 (P = .04). Thus, delayed treatment with TFPI improves survival in septic rabbits.

Topics: Animals; Anti-Bacterial Agents; Anticoagulants; Blood Coagulation; Blood Pressure; Dose-Response Relationship, Drug; Drug Therapy, Combination; Escherichia coli Infections; Gentamicins; Humans; Lipoproteins; Oxygen; Peritonitis; Rabbits; Recombinant Proteins; Shock, Septic; Thromboplastin

The expression of a range of surface molecules/receptors that are important in the host response to infection and foreign antigens was examined using peritoneal macrophages isolated from patients on continuous ambulatory peritoneal dialysis (CAPD) with peritonitis. The macrophage phenotypic profile was compared with that of normal peripheral blood monocytes. Consistently there was increased expression by macrophages of CD14, ICAM-1 (CD54), Fc gamma RI (CD64), Fc gamma RII (CDw32), Fc gamma RIII (CD16), transferrin receptors (CD71) and tissue factor. Increased expression of MHC class II was marginally significant. There was no detectable expression of either the p55 (CD25) or p70 chains of the IL-2 receptor. The expression of the complement receptors, CR1 (CD35) and CR3 (CD11b, CD18), was reduced. The activity of well-known inflammatory cytokines, rather than uraemic molecules, can account for the phenotypic profile of these extravasated peritoneal macrophages. The results of this study indicate that peritoneal macrophages from CAPD patients with peritonitis display a phenotype consistent with them being in vivo-derived inflammatory macrophages, and that they are appropriate for use in studies of anti-inflammatory agents.

Topics: Antigens, CD; Biomarkers; Flow Cytometry; Histocompatibility Antigens Class I; Histocompatibility Antigens Class II; Humans; Immunoglobulin G; Immunophenotyping; Macrophages; Monocytes; Peritoneal Cavity; Peritonitis; Receptors, Complement; Receptors, Fc; Receptors, Interleukin-2; Thromboplastin

Endotoxin levels and mononuclear phagocyte thromboplastin activities in samples from peripheral blood and peritoneal fluid were determined in nine patients with secondary bacterial peritonitis (appendicitis with perforation, or diverticulitis) and in five control patients (uncomplicated duodenal ulcer or gallbladder stones). None or only negligible amounts of endotoxin, always less than 0.01 ng/dl (contamination), and no growth of bacteria were detected in controls. In the patients with peritonitis, peritoneal fluid samples always contained gram-negative bacteria, and large amounts (mean, 31.6 ng/dl) of endotoxin were seen. Plasma from these patients also contained endotoxin (mean, 0.56 ng/dl) despite negative blood cultures. Mononuclear phagocytes from controls had low thromboplastin values, whereas both circulating monocytes and peritoneal macrophages from peritonitis patients showed a substantial increase (multifold) of thromboplastin.

Topics: Adolescent; Adult; Aged; Ascitic Fluid; Bacterial Infections; Endotoxins; Female; Humans; Macrophages; Male; Middle Aged; Monocytes; Peritonitis; Phagocytes; Thromboplastin

Thromboplastin values in blood monocytes, peritoneal macrophages, and in tissue samples from lung, aortic wall, liver, spleen, pancreas, kidney, jejunum, and colon were determined at 4, 10, and 16 h after induction of acute peritonitis (cecal perforation) or sham operation in rats. A maximum 35-fold and 100-fold rise of values was respectively demonstrated in monocytes and peritoneal macrophages in septic animals as compared to controls. This monocyte-macrophage-derived thromboplastin is probably central to the activation of blood coagulation and fibrin depositions/adhesion formation in septic peritonitis. A simultaneous and significant fall in thromboplastin content of standardized specimens from lung, aortic wall, liver, spleen, pancreas, and jejunum was observed in rats with peritonitis. This could reflect mobilization of thromboplastin in favor of the infectious focus. No significant changes were detected in tissue from kidney, whereas samples from the colon of septic animals showed a consistent increase as compared to controls.

Topics: Animals; Macrophages; Male; Monocytes; Peritonitis; Rats; Rats, Inbred Strains; Thromboplastin; Tissue Distribution