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Protease-activated Receptor 2 is Associated with Activation of Human Macrophage Cell Line THP-1

  • Kang, Chon-Sik (Department of Oriental Pharmacy, College of Pharmacy, Wonkwang University) ;
  • Tae, Jin (Department of Oriental Pharmacy, College of Pharmacy, Wonkwang University) ;
  • Lee, Young-Mi (Department of Oriental Pharmacy, College of Pharmacy, Wonkwang University) ;
  • Kim, Byeong-Soo (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Moon, Woo-Sung (Departments of Pathology, Chonguk National University Medical School) ;
  • Kim, Dae-Ki (Departments of Immunology Chonbuk National University Medical School)
  • Published : 2005.12.30
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Abstract

Background: Protease-activated receptor 2 (PAR2) belongs to a family of G protein coupled receptors activated by proteolytic cleavage. Trypsin-like serine proteases interact with PAR2 expressed by a variety of tissues and immune cells. The aim of our study was to investigate whether PAR2 stimulation can lead to the activation of human mac rophages. Methods: PAR2-mediated proliferation of human macrophage cell line THP-1 was measured with MTT assay. We also examined the extracellular regulated kinase (ERK) phosphorylation and cytokine production induced by trypsin and PAR2-agonist using western blot and enzyme-linked immunosorbent assay (ELISA), respectively. Results: Treatment of trypsin or PAR2-activating peptide increased cell proliferation in a dose-dependent manner, and induced the activation of ERK1/2 in THP-1 cells. In addition, trypsin-induced cell proliferation was inhibited by pretreatment of an ERK inhibitor (pD98059) or trypsin inhibitor (SBTI). Moreover, PAR2 activation by trypsin increased the secretion of TNF-${\alpha}$ in THP-1 cells. Conclusion: There results suggest that P AR2 activation by trypsin-like serine proteases can induce cell proliferation through the activation of ERK in human macrophage and that PAR2 may playa crucial role in the cell proliferation and cytokine secretion induced by trypsin-like serine proteases.

Keywords

References

  1. Roitt I, Brostoff J, Male D: Immunology (sixth edition); Mononuclear phagocytes in immune defence. p147-162, London, UK, Mosby, 2002
  2. Brass LF, Molino M: Protease-activated G protein-coupled receptors on human platelets and endothelial cells. Thromb Haemostasis 78;234-241, 1997
  3. Steinhoff M, Buddenkotte J, Shpacovitch V, Rattenholl A, Moormann C, Vergnolle N, Luger TA, Hollenberg MD: Proteinase-activated receptors: transducers of proteinase-mediated signaling in inflammation and immune response. Endocr Rev 26;1-43, 2005 https://doi.org/10.1210/er.2003-0025
  4. Coughlin SR: Thrombin signalling and protease-activated receptors. Nature 407;258-264, 2000 https://doi.org/10.1038/35025229
  5. Nystedt S, Emilsson K, Wahlestedt C, Sundelin J: Molecular cloning of a potential proteinase activated receptor. Proc Natl Acad Sci USA 91;9208-9212, 1994
  6. Steinhoff M, Vergnolle N, Young SH, Tognetto M, Amadesi S, Ennes HS, Trevisani M, Hollenberg MD, Wallace JL, Caughey GH, Mitchell SE, Williams LM, Geppetti P, Mayer EA, Bunnett NW: Agonists of proteinase-activated receptor 2 induce inflammation by a neurogenic mechanism. Nat Med 6;151-158, 2000 https://doi.org/10.1038/72247
  7. Koshikawa N, Nagashima Y, Miyagi Y, Mizushima H, Yanoma S, Yasumitsu H, Miyazaki K: Expression of trypsin in vascular endothelial cells. FEBS Lett 409;442-448, 1997 https://doi.org/10.1016/S0014-5793(97)00565-6
  8. Cottrell GS, Amadesi S, Grady EF, Bunnett NW: Trypsin IV, a novel agonist of protease-activated receptors 2 and 4. J Biol Chem 279;13532-13539, 2004 https://doi.org/10.1074/jbc.M312090200
  9. Kim JA, Choi SC, Yun KJ, Kim DK, Han MK, Seo GS, Yeom JJ, Kim TH, Nah YH, Lee YM: Expression of protease-activated receptor 2 in ulcerative colitis. Inflamm Bowel Dis 9;224-229, 2003 https://doi.org/10.1097/00054725-200307000-00002
  10. Nystedt S, Ramakrishnan V, Sundelin J: The proteinase-activated receptor 2 is induced by inflammatory mediators in human endothelial cells. Comparison with the thrombin receptor. J Biol Chem 271;14910-14915, 1996 https://doi.org/10.1074/jbc.271.25.14910
  11. Dery O, Corvera CU, Steinhoff M, Bunnett NW: Proteinaseactivated receptors: novel mechanisms of signaling by serine proteases. Am J Physiol 274;C1429-C1452, 1998 https://doi.org/10.1152/ajpcell.1998.274.6.C1429
  12. Vergnolle N: Proteinase-activated receptor-2-activating peptides induce leukocyte rolling, adhesion, and extravasation in vivo. J Immunol 163;5064-5069, 1999
  13. Lindner JR, Kahn ML, Coughlin SR, Sambrano GR, Schauble E, Bernstein D, Foy D, Hafezi-Moghadam A, Ley K: Delayed onset of inflammation in protease-activated receptor-2-deficient mice. J Immunol 165;6504-6510, 2000 https://doi.org/10.4049/jimmunol.165.11.6504
  14. Scarborough RM, Naughton MA, Teng W, Hung DT, Rose J, Vu TKH, Wheaton VI, Turck CW, Coughlin SR: Tethered ligand agonist peptides: structural requirements for thrombin receptor activation reveal mechanism of proteolytic unmasking of agonist function. J Biol Chem 267;13146-13149, 1992
  15. Howells GL, Macey MG, Chinni C, Hou L, Fox MT, Harriott P, Stone SR: Proteinase-activated receptor-2: expression by human neutrophils. J Cell Sci 110;881-887, 1997
  16. Kahn ML, Nakanishi-Matsui M, Shapiro MJ, Ishihara H, Coughlin SR: Protease-activated receptors 1 and 4 mediate activation of human platelets by thrombin. J Clin Invest 103;879-887, 1999 https://doi.org/10.1172/JCI6042
  17. Mari B, Guerin S, Far DF, Breitmayer JP, Belhacene N, Peyron JF, Rossi B, Auberger P: Thrombin and trypsin-induced $Ca^{2+}$ mobilization in human T cell lines through interaction with different protease-activated receptors. FASEB J 10;309-316, 1996 https://doi.org/10.1096/fasebj.10.2.8641564
  18. Colognato R, Slupsky JR, Jendrach M, Burysek L, Syrovets T, Simmet T: Differential expression and regulation of protease-activated receptors in human peripheral monocytes and monocyte-derived antigen-presenting cells. Blood 102;2645-2652, 2003 https://doi.org/10.1182/blood-2002-08-2497
  19. Ishihara H, Connolly AJ, Zeng D, Kahn ML, Zheng YW, Timmons C, Tram T, Coughlin SR: Protease-activated receptor 3 is a second thrombin receptor in humans. Nature 386;502-506, 1997 https://doi.org/10.1038/386502a0
  20. Cocks TM, Moffatt JD: Protease-activated receptors: sentries for inflammation?. Trends Pharmacol Sci 21:103-108, 2000 https://doi.org/10.1016/S0165-6147(99)01440-6
  21. Temkin V, Kantor B, Weg V, Hartman ML, Levi-Schaffer F: Tryptase activates the mitogen-activated protein kinase/activator protein-1 pathway in human peripheral blood eosinophils, causing cytokine production and release. J Immunol 169;2662-2669, 2002 https://doi.org/10.4049/jimmunol.169.5.2662
  22. Suk K, Cha S: Thrombin-induced interleukin-8 production and its regulation by interferon-${\gamma}$ and prostaglandin $E_{2}$ in human monocytic U937 cells. Immunol Lett 67;223-227, 1999 https://doi.org/10.1016/S0165-2478(99)00015-2
  23. Johnson JD, Chang JP: Function and agonistspecific $Ca^{2+}$ signalling: the requirement for and mechanism of spatial and temporal complexity in $Ca^{2+}$ signals. Biochem Cell Biol 78;217-240, 2000 https://doi.org/10.1139/bcb-78-3-217
  24. Ando T, Homma R, Ino Y, Ito G, Miyahara A, Yanagihara T, Kimura H, Ikeda S, Yamakawa H, Iwaki M: Trypsin-like protease of mites: purification and characterization of trypsin-like protease from mite faecal extract Dermatophagoides farinae: relationship between trypsin-like protease and Der f III. Clin Exp Allergy 23;777-784, 1993 https://doi.org/10.1111/j.1365-2222.1993.tb00366.x
  25. Schwartz LB, Lewis RA, Austen KF: Tryptase from human pulmonary mast cells: purification and characterization. J Biol Chem 256;11939-11943, 1981