Regulation of Murine Protein C Gene Expression In Vivo: Effects of Tumor Necrosis Factor-α, Interleukin-1, and Transforming Growth Factor-β

Summary

Protein C is a precursor of the anticoagulant serine protease, activated protein C, which inhibits coagulation factors Va and VIIIa. Although the liver appears to be the primary site of protein C synthesis, we previously demonstrated that the kidney and male reproductive organs also expressed abundant protein C mRNA in the mouse. In the present study, we further investigated the effects of tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), and transforming growth factor-β (TGF-β) on the expression of protein C mRNA in the principal producing organs, i.e., the liver, kidney, and testis. Both quantitative reverse transcription-PCR assay and in situ hybridization analysis revealed that TNF-α decreased protein C mRNA expression in the liver, kidney, and testis. IL-1 also down-regulated protein C mRNA expression in the liver and testis, but not in the kidney. In contrast, TGF-β unchanged the expression level of protein C mRNA in these three organs. These observations suggest that TNF-α and IL-1 may contribute to an increase in the procoagulant potential by down-regulation of protein C synthesis in the tissues during inflammatory processes.

• References

• 1 Kisiel W. Human plasma protein C. Isolation, characterization and mechanism of activation by α-thrombin. J Clin Invest 1979; 64: 761-9.
• 2 Esmon CT. The roles of protein C and thrombomodulin in the regulation of blood coagulation. J Biol Chem 1989; 264: 4743-6.
• 3 Marlar RA, Kleiss AJ, Griffin JH. Mechanism of action of human activated protein C, a thrombin-dependent anticoagulant enzyme. Blood 1982; 59: 1067-72.
• 4 van Hinsbergh VWM, Bertina RM, van Wijngaarden A, van Tilburg NH, Emeiss JJ, Haverkate F. Activated protein C decreases plasminogen activator-inhibitor activity in endothelial cell conditioned medium. Blood 1985; 65: 444-51.
• 5 Sakata Y, Loskutoff DJ, Gladson CL, Hekman CM, Griffin JH. Mechanism of protein C-dependent clot lysis: role of plasminogen activator inhibitor. Blood 1986; 68: 1218-23.
• 6 Bajzar L, Manuel R, Nesheim ME. Purification and characterization of TAFI, a thrombin-activatable fibrinolysis inhibitor. J Biol Chem 1995; 270: 14477-84.
• 7 Seligsohn U, Berger A, Abend M, Rubin L, Attias D, Zivelin A, Rapaport SI. Homozygous protein C deficiency manifested by massive venous thrombosis in the newborn. N Engl J Med 1984; 310: 559-62.
• 8 Bovill EG, Bauer KA, Dickerman JD, Callas P, West B. The clinical spectrum of heterozygous protein C deficiency in a large new England kindred. Blood 1989; 73: 712-7.
• 9 Kurosawa-Ohsawa K, Kimura M, Kume-Iwaki A, Tanaka T, Tanaka S. Anti-protein C monoclonal antibody induces thrombus in mice. Blood 1990; 75: 2156-63.
• 10 Taylor FB, Chang Jr A, Esmon CT, D’Angelo A, Vigano-D’Angelo S, Blick KE. Protein C prevents the coagulopathic and lethal effects of Escherichia coli infusion in the baboon. J Clin Invest 1987; 79: 918-25.
• 11 Gerson WT, Dickerman JD, Bovill EG, Golden E. Severe acquired protein C deficiency in purpura fulminans associated with disseminated intravascular coagulation: treatment with protein C concentrate. Pediatrics 1993; 91: 418-22.
• 12 Beckmann RJ, Shmidt RJ, Santerre RF, Plutzky J, Crabtree GR, Long GL. The structure and evolution of a 461 amino acid human protein C precursor and its messenger RNA, based upon the DNA sequence of cloned human liver cDNAs. Nucleic Acids Res 1985; 13: 5233-47.
• 13 Okafuji T, Maekawa K, Nawa K, Marumoto Y. The cDNA cloning and mRNA expression of rat protein C. Biochem Biophys Acta 1992; 1131: 329-32.
• 14 Jamison CS, McDowell SA, Marlar RA, Degen SJF. Developmental expression of protein C and protein S in the rat. Thromb Res 1995; 78: 407-19.
• 15 Yamamoto K, Loskutoff DJ. Extrahepatic expression and regulation of protein C in the mouse. Am J Pathol 1998; 153: 547-55.
• 16 He X, Shen L, Bjartell A, Malm J, Lilja H, Dahlback B. The gene encoding vitamin K-dependent anticoagulant protein C is expressed in human male reproductive tissues. J Histochem Cytochem 1995; 43: 563-70.
• 17 Esmon CT. The regulation of natural anticoagulant pathways. Science 1987; 235: 1348-52.
• 18 He X, Shen L, Bjartell A, Dahlback B. The gene encoding vitamin K-dependent anticoagulant protein S is expressed in multiple rabbit tissues as demonstrated by Northern blotting, immunocytochemistry and in situ hybridization. J Histochem Cytochem 1995; 43: 85-96.
• 19 Radtke K-P, Fernandez JA, Greengard JS, Tang WW, Wilson CB, Loskutoff DJ. Protein C inhibitor is expressed in tubular cells of human kidney. J Clin Invest 1994; 94: 2117-24.
• 20 Malm J, He X, Bjartell A, Shen L, Dahlback B. Synthesis of vitamin K-dependent protein S in the Leidig cells of human testis. Biochem J 1994; 302: 845-50.
• 21 Laurell M, Christensson A, Abrahamsson P-A, Stenflo J, Lija H. Protein C inhibitor in human body fluids. Seminal plasma is rich in inhibitor antigen deriving from cells throughout the male reproductive system. J Clin Invest 1992; 89: 1094-101.
• 22 Scarpati EM, Sadler JE. Regulation of endothelial cell coagulant properties: Modulation of tissue factor, plasminogen activator inhibitors, and thrombomodulin by phorbol 12-myristate 13-acetate and tumor necrosis factor. J Biol Chem 1989; 264: 20705-13.
• 23 Schleef RR, Bevilacqua MP, Sawdey M, Gimbrone Jr MA, Loskutoff DJ. Cytokine activation of vascular endothelium. Effects on tissue-type plasminogen activator and type-1 plasminogen activator inhibitor. J Biol Chem 1988; 263: 5797-803.
• 24 Sawdey M, Podor TJ, Loskutoff DJ. Regulation of type 1 plasminogen activator inhibitor gene expression in cultured bovine aortic endothelial cells: induction by transforming growth factor-β, lipopolysaccharide, and tumor necrosis factor-α . J Biol Chem 1989; 264: 10396-401.
• 25 Idell S, Kumar A, Zwieb C, Holiday D, Koenig KB, Johnson AR. Effects of TGF-β and TNF-α on procoagulant and fibrinolytic pathways of human tracheal epithelial cells. Am J Physiol 1994; 267: 693-703.
• 26 Sawdey MS, Loskutoff DJ. Regulation of murine type 1 plasminogen activator inhibitor gene expression in vivo. Tissue specificity and induction by lipopolysaccharide, tumor necrosis factor-α, and transforming growth factor-β. J Clin Invest 1991; 88: 1346-53.
• 27 Jansen PM, Boermeester MA, Fischer E, de Jong IW, van der Poll T, Moldawer LL, Hack CE, Lowry SF. Contribution of interleukin-1 to activation of coagulation and fibrinolysis, neutrophil degranulation, and the release of secretory-type phospholipase A₂ in sepsis: Studies in nonhuman primates after interleukin-1α administration and during lethal bacteremia. Blood 1995; 86: 1027-34.
• 28 Samad F, Yamamoto K, Pandey M, Loskutoff DJ. Elevated expression of transforming growth factor-β in adipose tissue from obese mice. Mol Med 1997; 3: 37-48.
• 29 Boswell JM, Yui MA, Burt DW, Delley VE. Increased tumor necrosis factor and IL-1β gene expression in the kidneys of mice with lupus nephritis. J Immunol 1988; 141: 3050-4.
• 30 Brennan DC, Yui MA, Wuthrich RP, Kelley VE. Tumor necrosis factor and IL-1 in New Zealand Black/White mice. Enhanced gene expression and acceleration of renal injury. J Immunol 1989; 143: 3470-5.
• 31 Hotamisligil GS, Shargill NS, Spiegelman BM. Adipose expression of tumor necrosis factor-α: Direct role in obesity-linked insulin resistance. Science 1993; 259: 87-91.
• 32 Yamamoto K, Loskutoff DJ. Fibrin deposition in tissues from endotoxin-treated mice correlates with decreases in the expression of urokinase-type but not tissue-type plasminogen activator. J Clin Invest 1996; 97: 2440-51.
• 33 Tada N, Sato M, Tsujimura A, Iwase R, Hashimoto-Gotoh T. Isolation and characterization of a mouse protein C cDNA. J Biochem 1992; 111: 491-5.
• 34 Tokunaga K, Taniguchi H, Yoda K, Shimizu M, Sakiyama S. Nucleotide sequence of a full-length cDNA for mouse cytoskeletal β-actin mRNA. Nucleic Acids Res 1986; 14: 2829.
• 35 Keeton M, Eguchi Y, Sawdey M, Ahn C, Loskutoff DJ. Cellular localization of type 1 plasminogen activator inhibitor messenger RNA and protein in murine renal tissue. Am J Pathol 1993; 142: 59-70.
• 36 Griffin J, Mosher D, Zimmerman T, Kleiss A. Protein C, an antithrombotic protein, is reduced in hospitalized patients with intravascular coagulation. Blood 1982; 60: 261-4.
• 37 Marlar RA, Endres-Brooks J, Miller C. Serial studies of protein C and its plasma inhibitor in patients with disseminated intravascular coagulation. Blood 1985; 66: 59-63.
• 38 Fijnvandraat K, Derkx B, Peters M, Bijlmer R, Sturk A, Prins MH, van Deventer SJH, ten Cate JW. Coagulation activation and tissue necrosis in meningococcal septic shock: severly reduced protein C levels predict a high mortality. Thromb Haemost 1995; 73: 15-20.
• 39 Nawroth PP, Stern DM. Modulation of endothelial cell hemostatic properties by tumor necrosis factor. J Exp Med 1986; 163: 740-5.
• 40 Dinarello CA. Interleukin-1 and interleukin-1 antagonism. Blood 1991; 77: 1627-52.
• 41 Emeis JJ, Kooistra T. Interleukin 1 and lipopolysaccharide induce an inhibitor of tissue-type plasminogen activator in vivo and in cultured endothelial cells. J Exp Med 1986; 163: 1260-6.
• 42 Bevilacqua MP, Schleef RR, Gimbrone Jr MA, Loskutoff DJ. Regulation of the fibrinolytic system of cultured vascular endothelium by interleukin 1. J Clin Invest 1986; 78: 587-91.
• 43 Conway EM, Rosenberg RD. Tumor necrosis factor suppresses transcription of the thrombomodulin gene in endothelial cells. Mol Cell Biol 1988; 8: 5588-92.
• 44 Lentz SR, Tsiang M, Sadler JE. Regulation of thrombomodulin by tumor necrosis factor-α: Comparison of transcriptional and posttranscriptional mechanisms. Blood 1991; 77: 542-50.
• 45 Nawroth PP, Handley DA, Esmon CT, Stern DM. Interleukin 1 induces endothelial cell procoagulant while suppressing cell-surface anticoagulant activity. Proc Natl Acad Sci USA 1986; 83: 3460-4.
• 46 Fischer E, Marano MA, Barber AE, Hudson A, Lee K, Rock CS, Hawes AS, Thompson RC, Lowry SF, Moldawer LL. Comparison between effects of interleukin-1α administration and sublethal endotoxemia in primates. Am J Pathol 1991; 261: 442-52.
• 47 Cybulsky MI, Chan MKW, Movat HZ. Biology of disease. Acute inflammation and microthrombosis induced by endotoxin, interleukin-1, and tumor necrosis factor and their implication in Gram-negative infection. Lab Invest 1988; 58: 365-78.
• 48 Kant KS, Pollak VE, Weiss MA, Glueck HI, Miller MA, Hess EV. Glomerular thrombosis in systemic lupus erythematosus: Prevalence and significance. Medicine 1981; 60: 71-86.
• 49 Viberti G, Wiseman MJ, Pinto JR, Messent J. Diabetic nephropathy. In: Kahn CR, Weir GC. editors Joslin’s Diabetes Mellitus. Philadelphia: Lea & Febiger; 1994: 691-737.
• 50 Moll S, Menoud PA, Fulpius T, Pastore Y, Takahashi S, Fossati L, Vassalli JD, Sappino AP, Schifferli JA, Izui S. Induction of plasminogen activator inhibitor type 1 in murine lupus-like glomerulonephritis. Kidney Int 1995; 48: 1459-68.
• 51 Yamamoto T, Noble NA, Cohen AH, Nast CC, Hishida A, Gold LI. et al Expression of transforming growth factor β is elevated in human and experimental diabetic nephropathy. Proc Natl Acad Sci USA 1993; 90: 1814-8.