Stabilization of monocyte chemoattractant protein-1-mRNA by activated protein C

Martina Brueckmann; Antje Marx; Hans Martin Weiler; Volker Liebe; Siegfried Lang; Jens J. Kaden; Wolfgang Zieger; Martin Borggrefe; Guenter Huhle; Karl Konstantin Haase

doi:10.1055/s-0037-1613554

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2003; 89(01): 149-160
DOI: 10.1055/s-0037-1613554

Wound Healing and Inflammation/Infection

Schattauer GmbH

Stabilization of monocyte chemoattractant protein-1-mRNA by activated protein C

Authors

Martina Brueckmann

¹1^st Department of Medicine, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Mannheim, Germany
Antje Marx

¹1^st Department of Medicine, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Mannheim, Germany
Hans Martin Weiler

¹1^st Department of Medicine, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Mannheim, Germany
Volker Liebe

¹1^st Department of Medicine, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Mannheim, Germany
Siegfried Lang

¹1^st Department of Medicine, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Mannheim, Germany
Jens J. Kaden

¹1^st Department of Medicine, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Mannheim, Germany
Wolfgang Zieger

²Department of Gynecology, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Mannheim, Germany
Martin Borggrefe

¹1^st Department of Medicine, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Mannheim, Germany
Guenter Huhle

¹1^st Department of Medicine, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Mannheim, Germany
Karl Konstantin Haase

¹1^st Department of Medicine, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Mannheim, Germany

Abstract

Summary

The activated protein C (APC) pathway has been suggested to be a common link between coagulation and inflammation. APC may function to restore hemostasis via modulation of cytokine expression. We investigated the effect of APC on the endothelial expression of monocyte chemoattractant protein-1 (MCP-1), a chemokine that is controlled by the activation of central proinflammatory transcription factors, such as nuclear factor-kappa B (NF-κ B).

We found that human APC (2.5-10 μ g/ml) upregulated the amount of MCP-1-mRNA in human umbilical vein endothelial cells (HUVEC) and caused a time- and dose-dependent increase in MCP-1 protein production (p <0.001 for APC 2.5 μg/ ml at 4 up to 24 h). In this cell culture model MCP-1 induced an improvement of cell migration and wound repair after injury to endothelial monolayers. After stimulation of MCP-1-mRNA-transcription with TNF-α (0.1-1 ng/ml), HUVEC’s were washed and an inhibitor of gene transcription, Actinomycin D (1 μg/ml), was added in the presence or absence of APC. HUVEC’s receiving APC contained more MCP-1-mRNA than controls after one hour and up to eight hours suggesting an inhibitory effect of APC on MCP-1-mRNA degradation (with APC: 753 ± 56 atto mol of MCP-1-mRNA per ml of cell lysate vs. 263 ± 60 atto mol/ml without APC at t =4 h; p <0.001). Electrophoretic mobility shift assays revealed that APC attenuated NF- κB DNA-binding capacity implying that NF- B may not be involved in the upregulatory effect of APC on MCP-1 production.

The ability of APC to upregulate the production of MCP-1, most likely by increasing the stability of MCP-1-mRNA rather than by transcriptional activation via NF- B, identifies a novel immunomodulatory pathway, by which APC may control the local inflammatory reaction, thereby initiating wound repair and modulating the extent of endothelial injury.

Keywords

Endothelium - activated protein C - monocyte chemoattractant protein-1 - mRNA-stability - wound repair

Volltext

Referenzen

References
1 Esmon CT. The protein C anticoagulant pathway. Arterioscler Thromb 1992; 12: 135-45.
2 Esmon CT, Taylor FB, Snow RT. Inflammation and coagulation: linked processes potentially regulated through a common pathway mediated by protein C. Thromb Haemost 1991; 66: 160-5.
3 Yan SB, Grinnell BW. Recombinant human protein C, protein S, and thrombomodulin as antithrombotics. Perspect Drug Discovery Res 1993; 1: 503-20.
4 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-gamma, or phorbol ester. J Immunol 1994; 153: 3664-72.
5 Schmidt-Supprian M, Murphy C, White B, Lawler M, Kapurniotu A, Voelter W, Smith O, Berhagen J. Activated protein C inhibits tumor necrosis factor and magrophage migration inhibitory factor production in monocytes. Eur Cytokine Netw 2000; 11: 407-13.
6 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.
7 Hooper WC, Phillips DJ, Renshaw MA, Evatt BL, Benson JM. The up-regulation of IL-6 and IL-8 in human endothelial cells by activated protein C. J Immunol 1998; 161: 2567-73.
8 Rollins BJ. Monocyte chemoattractant protein-1: a potent regulator of monocyte recruitment in inflammatory disease. Mol Med Today 1996; 2: 198-204.
9 Carr MW, Roth SJ, Luther E, Rose SS, Springer TA. Monocyte chemoattractant protein-1 acts as a T-lymphocyte chemoattractant. Proc Natl Acad Sci USA 1994; 91: 3652-6.
10 Wang JM, Sica A, Peri G, Walter S, Padura IM, Libby P, Ceska M, Lindley I, Colotta E, Mantovani A. Expression of monocyte chemo-tactic protein and interleukin-8 by cytokine activated human vascular smooth muscle cells. Arterioscler Thromb Vasc Biol 1991; 11: 1166-74.
11 Brown Z, Gerritsen ME, Carley WW, Strieter RM, Kunkel SL, Westwick J. Chemokine gene expression and secretion by cytokine-activated human microvascular endothelial cells: differential regulation on monocyte chemoattractant protein-1 and interleukin 8 in response to interferon-gamma. Am J Pathol 1994; 145: 913-21.
12 Martin T, Cardarelli PM, Parry GC, Felts KA, Cobb RR. Cytokine induction of monocyte chemoattractant protein-1 gene expression in human endothelial cells depends on the cooperative action of NF-B and AP-1. Eur J Immunol 1997; 27: 1091-7.
13 Weber KSC, Nelson PJ, Groene H-J, Weber C. Expression of CCR2 by endothelial cells – implications for MCP-1 mediated wound injury repair and in vivo inflammatory activation of endothelium. Arterioscler Thromb Vasc Biol 1999; 19: 2085-93.
14 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins: identification by morphologic and immunologic criteria. J Clin Invest 1973; 52: 2745-56.
15 Dignam JD, Lebovitz RM, Roeder RG. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res 1983; 11: 1475-89.
16 Esmon CT, Esmon NL, LeBonnice BF, Johnson AE. Protein C activation. Methods Enzymol 1993; 222: 358.
17 Lowe HC, Fahmy RG, Kavurma MM, Baker A, Chesterman CN, Khachigian LM. Catalytic oligonucleotides define a key regulatory role for early growth response factor-1 in the porcine model of coronary in-stent restenosis. Circ Res 2001; 89: 670-7.
18 Lo LW, Cheng JJ, Chiu JJ, Wung BS, Liu YC, Wang DL. Endothelial exposure to hypoxia induces EGR-1 expression involving PKC alpha-mediated Ras/Raf-1/ERK1/2 pathway. J Cell Physiol 2001; 188: 304-12.
19 Teruyama K, Abe M, Nakano T, Iwasaka-Yagi C, Takahashi S, Yamada S, Sato Y. Role of transcription factor Ets-1 in the apoptosis of human vascular endothelial cells. J Cell Physiol 2001; 188: 243-52.
20 Esmon CT. The anticoagulant and anti-inflammatory roles of the protein C anticoagulant pathway. J Autoimmun 2000; 15: 113-6.
21 Ylä-Herttula S, Lipton BA, Sarkioja ME, Yoshimura T, Leonard EJ, Witztum JL, Steinberg D. Expression of monocyte chemotactic protein-1 in macrophage-rich areas of human and rabbit atherosclerotic lesions. Proc Natl Acad Sci USA 1991; 88: 5252-6.
22 Zisman DA, Kunkel SL, Strieter RM, Tsai WC, Bucknell K, Wilkowski J, Standiford TJ. MCP-1 protects mice in lethal endotoxemia. J Clin Invest 1997; 99: 2832-6.
23 Matsukawa A, Hogaboam CM, Lukacs NW, Lincoln PM, Strieter RM, Kunkel SL. Endogenous MCP-1 influences systemic cytokine balance in a murine model of acute septic peritonitis. Exp Mol Pathol 2000; 68: 77-84.
24 Hooper WC, Phillips DJ, Renshaw MA. Activated protein C induction of MCP-1 in human endothelial cells – a possible role for endothelial cell nitric oxide synthase. Thrombosis Research 2001; 103: 209-19.
25 Ueda A, Okuda K, Ohno S, Shirai A, Igarashi T, Matsunaga K, Fukushima J, Kawamoto S, Ishigatsubo Y, Okubo T. NF-kappa B and Sp1 regulate transcription of the human monocyte chemoattractant gene. J Immunol 1994; 153: 2052-63.
26 Barnes PF, Karin M. Nuclear factor-kappa B: a pivotal transcription factor in chronic inflammatory diseases. New Engl J Med 1997; 336: 1066-71.
27 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 on nuclear factor κB (NF-B) and tumor necrosis factor a (TNF-) production in the THP-1 monocytic cell line. Br J Haematol 2000; 110: 130-4.
28 Stula M, Orzechowski HD, Gschwend S, Vetter R, von Harsdorf R, Dietz R, Paul M. Influence of sustained mechanical stress on EGR-1 mRNA expression in cultured human endothelial cells. Mol Cell Biochem 2000; 210: 101-8.
29 Bochkov VN, Mechtcheriakova D, Lucerna M, Huer J, Malli R, Graier WF, Hofer E, Binder BR, Leitinger N. Oxidized phospho-lipids stimulate tissue factor expression in human endothelial cells via activation of ERK/EGR-1 and Ca(++)/NFAT. Blood 2002; 99: 199-206.