Tissue Factor Pathway Inhibitor Production by Human Mesangial Cells in Culture

Summary

Fibrin formation within the glomeruli has been observed in various forms of human and experimental glomerulonephritis and it may play an important role in progressive glomerular injury. Furthermore it has been hypothesized that glomerular fibrin deposition may occur through activation of either the intrinsic or extrinsic coagulation pathway. It has been demonstrated that a procoagulant activity (PCA) which is compatible with tissue factor is present in the glomeruli and becomes increased in human proliferative glomerulonephritis and in animal models of nephritis. Tissue factor pathway inhibitor (TFPI) regulates the extrinsic pathway of blood coagulation through its ability to inhibit tissue factor activity. TFPI is present in plasma and in platelets, and it is now thought to be produced mainly by endothelial cells. We examined whether human mesangial cells (HMC) could produce TFPI and attempted to clarify regulatory factors which affect TFPI production. Cultured HMC were used and TFPI in the cell supernatants was measured by ELISA using a specific antibody. Cultured HMC showed the production of TFPI. Immunoblot analysis revealed 40 kD protein of TFPI. The concentration of TFPI was significantly increased following the incubation with thrombin and heparin, including low molecular weight heparin, in a dose- and time-dependent manner. However, fetal calf serum, phorbol myristate acetate, lipopolysaccharide, IL-1β and tissue factor did not stimulate TFPI synthesis. Our data show that cultured HMC have the ability to produce TFPI which inhibits fibrin formation. It is possible that thrombin-induced enhancement of TFPI synthesis may be caused by the autoregulatory system of blood coagulation and that with heparin it may represent another anticoagulatory effect of heparin.

• References

• 1 Tipping PG, Dowling JP, Holdsworth SR. Glomerular procoagulant activity in human proliferative glomerulonephritis. J Clin Invest 1988; 81: 119-125
• 2 Holdsworth SR, Tipping PG. Macrophage-induced glomerular fibrin deposition in experimental glomerulonephritis in the rabbit. J Clin Invest 1985; 76: 1367-1374
• 3 Tipping PG, Holdsworth SR. The participation of macrophages, glomerular procoagulant activity, and factor VIII in glomerular fibrin deposition. Am J Pathol 1986; 124: 10-17
• 4 Wiggins RC, Glatfelter A, Brukman J. Procoagulant activity in glomeruli and urine of rabbits with nephrotoxic nephritis. Lab Invest 1985; 53: 156-164
• 5 Wiggins RC, Njoku N, Sedor JR. Tissue factor production by cultured rat mesangial cells. Stimulation by TNF-α and lipopolysaccharide Kidney Int 1990; 37: 1281-1285
• 6 Yamabe H, Yoshikawa S, Ohsawa H, Inuma H, Miyata M, Sasaki T, Kai-zuka M, Tamura N, Onodera K. Tissue factor production by cultured rat glomerular epithelial cells. Nephrol Dial Transplant 1993; 8: 519-523
• 7 Rapaport SI. The extrinsic pathway inhibitor: A regulator of tissue factor-dependent blood coagulation. Thromb Haemost 1991; 66: 6-15
• 8 Bajaj MS, Rana SV, Wysolmerski RB, Bajaj SP. Inhibitor of the factor VIIa-tissue factor complex is reduced in patients with disseminated intra-vascular coagulation but not in patients with severe hepatocellular disease. J Clin Invest 1987; 79: 1874-1878
• 9 Striker GE, Striker LJ. Biology of disease; glomerular cell culture. Lab Invest 1985; 53: 122-131
• 10 Broze Jr GJ, Girard TJ, Novotny WF. Regulation by a multivalent Kunitz-type inhibitor. Biochemistry 1990; 29: 7539-7546
• 11 Bajaj MS, Kuppuswamy MN, Saito H, Spitzer SG, Bajaj SP. Cultured normal human hepatocytes do not synthesize lipoprotein-associated coagulation inhibitor: Evidence that endothelium is the principle site of its synthesis. Proc Natl Acad Sci USA 1990; 87: 8869-8873
• 12 Werling RW, Zacharski LR, Kisiel W, Bajaj SP, Memoli VA, Rousseau SM. Distribution of tissue factor pathway inhibitor in normal and malignant human tissues. Thromb Haemost 1993; 69: 366-369
• 13 Tipping PG, Erlich JH, Apostolopoulos J, Holdsworth SR. Expression of tissue factor pathway inhibitor (TFPI) in fibrin dependent crescentic glomerulonephritis in rabbits. J Am Soc Nephrol 1994; 5: 796 (abstract)
• 14 Wam-Cramer Bj, Maki SL, Zivelin A, Rapaport SI. Partial purification and characterization of the extrisic pathway inhibitor (the factor Xa-dependent plasma inhibitor of factor Vila/tissue factor). Thromb Res 1987; 48: 11-22
• 15 Novotny WF. Tissue factor pathway inhibitor. Semin Thromb Haemost 1994; 20: 101-108
• 16 Villamediana LM, Rondeau E, He Cl, Medcalf RL, Peraldi MN, Lacave R, Delarue F, Sraer JD. Thrombin regulates components of the fibrinolytic system in human mesangial cells. Kidney Int 1990; 38: 956-961
• 17 Lacave R, Rondeau E, Ochi S, Delarue F, Schleuning WD, Spaer J-D. Characterization of plasminogen activator and its inhibitor in human mesangial cells. Kidney Int 1989; 35: 806-811
• 18 Ameri A, Kuppuswamy MN, Basu S, Bajaj SP. Expression of tissue factor pathway inhibitor by cultured endothelial cells in response to inflammatory mediators. Blood 1992; 79: 3219-3226
• 19 Van der Vogt CPE, Dirven RJ, Reitsma PH, Bertina RM. Expression of tissue factor and tissue factor pathway inhibitor in monocytes in response to bacterial lipopolysaccharide and phorbolester. Blood Coag Fibrinol 1994; 5: 211-220
• 20 Novotny WF, Girard TJ, Miletich JP, Broze Jr GJ. Platelets secrete a coagulation inhibitor functionally and antigenically similar to the lipoprotein associated coagulation inhibitor. Blood 1988; 72: 2020-2025
• 21 Sandset PM, Abildgaard U, Larsen ML. Heparin induces release of extrinsic coagulation pathway inhibitor (EPI). Thromb Res 1988; 50: 803-813
• 22 Kario K, Matsuo T, Yamada T, Matsuo M. Increased tissue factor pathway inhibitor levels in uremic patients on regular hemodialysis. Thromb Haemost 1994; 71: 275-279
• 23 Norrheim L, Abildgaard U, Larsen ML, Lindahl AK. Involvement of the extrinsic pathway in the activities of low molecular weight heparins. Thromb Res 1991; 14: 19-27
• 24 Abildgaard U. Heparin/low molecular weight heparin and tissue factor pathway inhibitor. Haemostasis 1993; 23: 103-106
• 25 Harenberg J, Siegele M, Dempfle CE, Stehle G, Heene DL. Protamine neutralization of the release of tissue factor pathway inhibitor activity by heparins. Thromb Haemost 1993; 70: 942-945