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Thromb Haemost 2002; 88(02): 267-273
DOI: 10.1055/s-0037-1613197
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Activated Protein C Inhibits Lipopolysaccharide-Induced Tumor Necrosis Factor-α Production by Inhibiting Activation of both Nuclear Factor-κB and Activator Protein-1 in Human Monocytes

Mehtap Yuksel
1   Department of Laboratory Medicine, Kumamoto, Japan
2   Department of Biochemistry, Kumamoto University School of Medicine, Kumamoto, Japan
,
Kenji Okajima
1   Department of Laboratory Medicine, Kumamoto, Japan
,
Mitsuhiro Uchiba
1   Department of Laboratory Medicine, Kumamoto, Japan
,
Seikoh Horiuchi
2   Department of Biochemistry, Kumamoto University School of Medicine, Kumamoto, Japan
,
Hiroaki Okabe
1   Department of Laboratory Medicine, Kumamoto, Japan
› Author Affiliations
Further Information

Publication History

Received 10 December 2001

Accepted after revision 11 April 2002

Publication Date:
27 December 2017 (online)

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Summary

Activated protein C (APC), an important natural anticoagulant, inhibits tumor necrosis factor-α (TNF-α) production and attenuates various deleterious events induced by lipopolysaccharide (LPS), contributing thereby to a significant reduction of mortality in patients with severe sepsis. In this study, we investigated the mechanism(s) by which APC inhibits TNF-α production by LPS-stimulated human monocytes in vitro. Although APC inhibited LPS-induced TNF-α production in a concentration-dependent fashion, diisopropyl fluorophosphate-treated APC, an active-site-blocked APC, had no effect. APC inhibited both the binding of nuclear factor-κB (NF-κB) to target sites and the degradation of IκBα. APC also inhibited both the binding of activator protein-1 (AP-1) to target sites and the activation of mitogen-activated protein kinase pathways. These observations strongly suggest that APC inhibited LPS-induced TNF-α production by inhibiting the activation of both NF-κB and AP-1 and that the inhibitory activity of APC might depend on its serine protease activity. These results would at least partly explain the mechanism(s) by which APC reduces the tissue injury seen in animal models of sepsis and in patients with sepsis.

Keywords

Activated protein C - human monocytes - TNF-α - NF-κB - AP-1
 

  • References

  • 1 Esmon CT. The protein C anticoagulant pathway. Arterioscler Thromb 1992; 12: 135-45.
    CrossrefPubMedGoogle Scholar
  • 2 Walker FJ, Sexton PW, Esmon CT. The inhibition of blood coagulation by activated protein C through the selective inactivation of activated factor V. Biochim Biophys Acta 1979; 571: 333-42.
    CrossrefPubMedGoogle Scholar
  • 3 Fulcher CA, Gardiner JE, Griffin JH, Zimmerman TS. Proteolytic inactivation of human factor VIII procoagulant protein by activated human protein C and its analogy with factor V. Blood 1984; 63: 486-9.
    PubMedGoogle Scholar
  • 4 Taylor Jr FB, Chang 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.
    CrossrefPubMedGoogle Scholar
  • 5 Murakami K, Okajima K, Uchiba M, Johno M, Nakagaki T, Okabe H, Takatsuki K. Activated protein C attenuates endotoxin-induced pulmonary vascular injury by inhibiting activated leukocytes in rats. Blood 1996; 87: 642-7.
    PubMedGoogle Scholar
  • 6 Isobe H, Okajima K, Uchiba M, Mizutani A, Harada N, Nagasaki A, Okabe K. Activated protein C prevents endotoxin-induced hypotension in rats by inhibiting excessive production of nitric oxide. Circulation 2001; 104: 1171-5.
    CrossrefPubMedGoogle Scholar
  • 7 Mizutani A, Okajima K, Uchiba M, Noguchi T. Activated protein C reduces ischemia/reperfusion-induced renal injury in rats by inhibiting leukocyte activation. Blood 2000; 95: 3781-7.
    PubMedGoogle Scholar
  • 8 Murakami K, Okajima K, Uchiba M, Johno M, Nakagaki T, Okabe H, Takatsuki K. Activated protein C prevents LPS-induced pulmonary vascular injury by inhibiting cytokine production. Am J Physiol 1997; 272: L197-L202.
    PubMedGoogle Scholar
  • 9 Levi M, de Jonge E, van der Poll T, ten Cate H. Disseminated intravascular coagulation. Thromb Haemost 1999; 82: 695-705.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 10 Bernard GR, Vincent JL, Laterre PF, LaRosa SP, Dhainaut JF, Lopez-Rodriguez A, Steingrub JS, Garber GE, Helterbrand JD, Ely EW, Fisher CJ. Efficacy and safety of recombinant human activated protein C for severe sepsis. N Engl J Med 2001; 344: 699-709.
    CrossrefPubMedGoogle Scholar
  • 11 Baldwin Jr AS. The NF-κB and IκB proteins: new discoveries and insights. Annu Rev Immunol 1996; 14: 649-81.
    CrossrefPubMedGoogle Scholar
  • 12 Karin M, Ben-Neriah Y. Phosphorylation meets ubiquitination: the control of NF-κB activity. Annu Rev Immunol 2000; 18: 621-63.
    CrossrefPubMedGoogle Scholar
  • 13 Finco TS, Baldwin AS. Mechanistic aspects of NF-κB regulation: the emerging role of phosphorylation and proteolysis. Immunity 1995; 03: 263-72.
    CrossrefPubMedGoogle Scholar
  • 14 Davis RJ. MAPKs: new JNK expands the group. Trends Biochem Sci 1994; 19: 470-3.
    CrossrefPubMedGoogle Scholar
  • 15 Hambleton J, Weinstein SL, Lem L, DeFranco AL. Activation of c-Jun N-terminal kinase in bacterial lipopolysaccharide-stimulated macrophages. Proc Natl Acad Sci USA 1996; 93: 2774-8.
    CrossrefPubMedGoogle Scholar
  • 16 Lee JC, Laydon JT, McDonnell PC, Gallagher TF, Kumar S, Green D, McNulty D, Blumenthal MJ, Heys JR, Landvatter SW, Strickler JE, McLaughlin MM, Siemens IR, Fisher SM, Livi GP, White JR, Adams JL, Young PR. A protein kinase involved in the regulation of inflammatory cytokine biosynthesis. Nature 1994; 372: 739-46.
    CrossrefPubMedGoogle Scholar
  • 17 Karin M. The regulation of AP-1 activity by mitogen-activated protein kinases. J Biol Chem 1995; 270: 16483-6.
    CrossrefPubMedGoogle Scholar
  • 18 Whitmarsh AJ, Davis RJ. Transcription factor AP-1 regulation by mitogenactivated protein kinase signal transduction pathways. J Mol Med 1996; 74: 589-607.
    CrossrefPubMedGoogle Scholar
  • 19 White B, Schmidt M, Murphy C, Livingstone W, O'Toole D, Lawler M, O'Neill L, Kelleher D, Schwarz HP, Smith OP. Activated protein C inhibits lipopolysaccharide-induced nuclear translocation of nuclear factor κB (NF-κB) and tumor necrosis factor α (TNF-α) production in the THP-1 monocytic cell line. Br J Haematol 2000; 110: 130-4.
    CrossrefPubMedGoogle Scholar
  • 20 Schmidt-Supprian M, Murphy C, White B, Lawler M, Kapurniotu A, Voelter W, Smith O, Bernhagen J. Activated protein C inhibits tumor necrosis factor and macrophage migration inhibitory factor production in monocytes. Eur Cytokine Netw 2000; 11: 407-13.
    PubMedGoogle Scholar
  • 21 Comp PC, Nixon RR, Esmon CT. Determination of functional levels of protein C, an antithrombotic protein, using thrombin-thrombomodulin complex. Blood 1984; 63: 115-21.
    PubMedGoogle Scholar
  • 22 Grey ST, Tsuchida A, Hau H, Orthner CL, Salem HH, Hancock WW. Selective inhibitory effects of the anticoagulant activated protein C on the responses of human mononuclear phagocytes to LPS, IFN-γ, or phorbol ester. J Immunol 1994; 153: 3664-72.
    PubMedGoogle Scholar
  • 23 Uchiba M, Okajima K, Murakami K, Okabe H, Takatsuki K. Effect of nafamostat mesilate on pulmonary vascular injury induced by lipopolysaccharide in rats. Am J Respir Crit Care Med 1997; 155: 711-8.
    CrossrefPubMedGoogle Scholar
  • 24 Cheshire JL, Baldwin Jr AS. Synergistic activation of NF-κB by tumor necrosis factor alpha and gamma interferon via enhanced IκBα degradation and de novo IκBβ degradation. Mol Cell Biol 1997; 17: 6746-54.
    CrossrefPubMedGoogle Scholar
  • 25 Yao J, Mackman N, Edgington TS, Fan ST. Lipopolysaccharide induction of the tumor necrosis factor-α promoter in human monocytic cells: regulation by Egr-1, c-Jun, and NF-κB transcription factors. J Biol Chem 1997; 272: 17795-801.
    CrossrefPubMedGoogle Scholar
  • 26 Karin M, Liu Z, Zandi E. AP-1 function and regulation. Curr Opin Cell Biol 1997; 09: 240-6.
    CrossrefPubMedGoogle Scholar
  • 27 Kyriakis JM, Banerjee P, Nikolakaki E, Dai T, Rubie EA, Ahmad MF, Avruch J, Woodgett JR. The stress-activated protein kinase subfamily of c-Jun kinases. Nature 1994; 369: 156-60.
    CrossrefPubMedGoogle Scholar
  • 28 Derijard B, Hibi M, Wu IH, Barrett T, Su B, Deng T, Karin M, Davis RJ. JNK1: a protein kinase stimulated by UV light and Ha-Ras that binds and phosphorylates the c-Jun activation domain. Cell 1994; 76: 1025-37.
    CrossrefPubMedGoogle Scholar
  • 29 Minden A, Karin M. Regulation and function of the JNK subgroup of MAP kinases. Biochim Biophys Acta 1997; 1333: F85-F104.
    PubMedGoogle Scholar
  • 30 Raingeaud J, Gupta S, Rogers JS, Dickens M, Han J, Ulevitch RJ, Davis RJ. Pro-inflammatory cytokines and environmental stress cause p38 mitogenactivated protein kinase activation by dual phosphorylation on tyrosine and threonine. J Biol Chem 1995; 270: 7420-6.
    CrossrefPubMedGoogle Scholar
  • 31 Steffan NM, Bren GD, Frantz B, Tocci MJ, O’Neill EA, Paya CV. Regulation of IκBα phosphorylation by PKC- and Ca2+-dependent signal trandusction pathways. J Immunol 1995; 155: 4685-91.
    PubMedGoogle Scholar
  • 32 Hancock WW, Grey ST, Hau L, Akalin E, Orthner C, Sayegh MH, Salem HH. Binding of activated protein C to a specific receptor on human mononuclear phagocytes inhibits intracellular calcium signaling and monocyte-dependent proliferative responses. Transplantation 1995; 60: 1525-32.
    CrossrefPubMedGoogle Scholar
  • 33 Chow CW, Grinstein S, Rotstein OD. Signaling events in monocytes and macrophages. New Horiz 1995; 03: 342-51.
    PubMedGoogle Scholar
  • 34 Carlos TM, Harlan JM. Leukocyte-endothelial adhesion molecules. Blood 1994; 84: 2068-101.
    PubMedGoogle Scholar
  • 35 Joyce DE, Gelbert L, Ciaccia A, DeHoff B, Grinnell BW. Gene expression profile of antithrombotic protein C defines new mechanisms modulating inflammation and apoptosis. J Biol Chem 2001; 276: 11199-203.
    CrossrefPubMedGoogle Scholar
  • 36 Stein B, Baldwin Jr AS, Ballard DW, Greene WC, Angel P, Herrlich P. Cross-coupling of the NF-κB p65 and Fos/Jun transcription factors produces potentiated biological function. EMBO J 1993; 12: 3879-91.
    PubMedGoogle Scholar
  • 37 Yasumoto K, Okamoto S, Mukaida N, Murakami S, Mai M, Matsushima K. Tumor necrosis factor α and interferon γ synergistically induce interleukin 8 production in a human gastric cancer cell line through acting concurrently on AP-1 and NF-κB-like binding sites of the interleukin 8 gene. J Biol Chem 1992; 267: 22506-11.
    PubMedGoogle Scholar
  • 38 Chen ZJ, Parent L, Maniatis T. Site-specific phosphorylation of IκBα by a novel ubiquitination-dependent protein kinase activity. Cell 1996; 84: 853-62.
    CrossrefPubMedGoogle Scholar
  • 39 Lee FS, Hagler J, Chen ZJ, Maniatis T. Activation of the IκBα kinase complex by MEKK1, a kinase of the JNK pathway. Cell 1997; 88: 213-22.
    CrossrefPubMedGoogle Scholar
  • 40 Derijard B, Raingeaud J, Barrett T, Wu IH, Han J, Ulevitch RJ, Davis RJ. Independent human MAP kinase signal transduction pathways defined by MEK and MKK isoforms. Science 1995; 267: 682-5.
    CrossrefPubMedGoogle Scholar
  • 41 Lin A, Minden A, Martinetto H, Claret FX, Lange-Carter C, Mercurio F, Johnson GL, Karin M. Identification of a dual specificity kinase that activates the Jun kinases and p38-Mpk2. Science 1995; 268: 286-90.
    CrossrefPubMedGoogle Scholar
  • 42 Taoka Y, Okajima K, Uchiba M, Murakami K, Harada N, Johno M, Naruo M. Activated protein C reduces the severity of compression-induced spinal cord injury in rats by inhibiting activation of leukocytes. J Neurosci 1998; 18: 1393-8.
    CrossrefPubMedGoogle Scholar
  • 43 Hirose K, Okajima K, Taoka Y, Uchiba M, Tagami H, Nakano K, Utoh J, Okabe H, Kitamura N. Activated protein C reduces the ischemia/reperfusioninduced spinal cord injury in rats by inhibiting neutrophil activation. Ann Surg 2000; 232: 272-80.
    CrossrefPubMedGoogle Scholar
  • 44 Yeh KY, Yeh M, Glass J, Granger DN. Rapid activation of NF-κB and AP-1 and target gene expression in postischemic rat intestine. Gastroenterology 2000; 118: 525-34.
    CrossrefPubMedGoogle Scholar
  • 45 Bone RC. The pathogenesis of sepsis. Ann Intern Med 1991; 115: 457-69.
    CrossrefPubMedGoogle Scholar
  • 46 Opal SM, Scannon PJ, Vincent JL, White M, Carroll SF, Palardy JE, Parejo NA, Pribble JP, Lemke JH. Relationship between plasma levels of lipopolysaccharide (LPS) and LPS-binding protein in patients with severe sepsis and septic shock. J Infect Dis 1999; 180: 1584-9.
    CrossrefPubMedGoogle Scholar
  • 47 Heeb MJ, Espana F, Griffin JH. Inhibition and complexation of activated protein C by two major inhibitors in plasma. Blood 1989; 73: 446-54.
    PubMedGoogle Scholar
  • 48 Marlar RA, Kressin DC, Madden RM. Contribution of plasma proteinase inhibitors to the regulation of activated protein C in plasma. Thromb Haemost 1993; 69: 16-20.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 49 Shu F, Kobayashi H, Fukudome K, Tsuneyoshi N, Kimoto M, Terao T. Activated protein C suppresses tissue factor expression on U937 cells in the endothelial protein C receptor-dependent manner. FEBS Lett 2000; 477: 208-12.
    CrossrefPubMedGoogle Scholar
  • 50 Taylor Jr FB, Stearns-Kurosawa DJ, Kurosawa S, Ferrell G, Chang ACK, Laszik Z, Kosanke S, Peer G, Esmon CT. The endothelial cell protein C receptor aids in host defense against Escherichia coli sepsis. Blood 2000; 95: 1680-6.
    PubMedGoogle Scholar
  • 51 Galligan L, Livingstone W, Volkov Y, Hokamp K, Murphy C, Lawler M, Fukudome K, Smith O. Characterization of protein C receptor expression in monocytes. Br J Haematol 2001; 115: 408-14.
    CrossrefPubMedGoogle Scholar