Korean Chemical Engineering Research

The Korean Institute of Chemical Engineers (한국화학공학회)
권호 표지

Effect of Soluble EPCR on the Anti-Inflammatory Effects by Activated Protein C

수용성 EPCR에 의한 활성화된 단백질 C의 항염증 작용에 관한 연구

  • Bae, Jong-Sup (Department of Herbal Pharmaceutical Engineering, Daegu Haany University) ;
  • Park, Moon-Ki (Department of Herbal Pharmaceutical Engineering, Daegu Haany University) ;
  • Park, Sang-Wook (Division of Chemical Engineering, Pusan National University)
  • 배종섭 (대구한의대학교 한방제약공학과) ;
  • 박문기 (대구한의대학교 한방제약공학과) ;
  • 박상욱 (부산대학교 화학공학과)
  • Received : 2009.03.25
  • Accepted : 2009.07.18
  • Published : 2009.08.31
Download PDF
⟨ Previous Next ⟩

Abstract

In this study, we evaluated the effect of soluble EPCR(Soluble Endothelial Protein C Receptor, sEPCR) on the anti-inflammatory activities by activated protein C(APC) in endothelium. We demonstrated that sEPCR inhibited the barrier protective activity, the inhibition of neutrophils adhesion toward endothelial cells and the inhibition of transendothelial migration by APC in endothelial cells. Interestingly, sEPCR also blocked the mechanism by which APC inhibited the expression of cell adhesion molecules(CAM) by TNF-alpha in endothelial cells. These results suggested that the anti-inflammatory activities of APC was inhibited by sEPCR which blocked the binding motifs of Gla domain of APC to membrane bound EPCR. This finding will provide the important evidence in the development of new medicine for the treatment of severe sepsis and inflammatory diseases and good clue for understanding unknown mechanisms by which APC showed the anti-inflammatory activities in endothelium.

본 논문에서는 혈관내피세포에서 활성화된 단백질 C(Activated Protein C, APC)의 항염증 작용에서 수용성 EPCR(Soluble Endothelial Protein C Receptor, sEPCR)의 효과를 관찰하였다. sEPCR은 APC가 매개하는 항염증 작용에 있어 내피세포막의 보호효과를 저해하고, 혈관내피세포에 대한 백혈구의 부착저해 효과를 억제하며, 혈관내피세포를 관통하는 백혈구의 이동을 저해하는 효과를 억제한다. 그리고 흥미롭게도 sEPCR은 내피세포에서 TNF-alpha에 의한 세포부착단백질의 발현을 억제하는 APC의 기전을 저해함으로써 APC가 가지는 항염증 효과를 억제한다. 이것은 APC의 Gla 도메인이 내피세포의 수용체인 EPCR에 결합할 수 있는 부위에 sEPCR이 상호작용함으로써 더 이상 APC이 세포막에 존재하는 EPCR과 결합을 못함으로써 APC의 항염증 작용은 억제되는 것을 의미한다. 이 결과는 향후 중증 패혈증 및 염증질환을 효과적으로 치료할 수 있는 신약개발에 중요한 단서를 제공할 것이고 내피세포에서 아직 명확하게 밝혀지지 않은 APC의 항염증 작용의 기전을 밝히는 데 좋은 정보를 제공할 것이다.

Keywords

Acknowledgement

Supported by : 농림부

References

  1. Esmon, C. T., "The Roles of Protein C and Thrombomodulin in the Regulation of Blood Coagulation," J. Biol. Chem., 264, 4743-4746(1989)
  2. Esmon, C. T., 'Thrombomodulin as a Model of Molecular Mechanisms that Modulate Protease Specificity and Function at the Vessel Surface,' FASEB J., 9(10), 946-955(1995) https://doi.org/10.1096/fasebj.9.10.7615164
  3. Esmon, C. T., 'Molecular Events that Control the Protein C Anticoagulant Pathway,' Thromb Haemostasis., 70, 29-35(1993)
  4. Stenflo, J., "Structure and Function of Protein C," Semin. Thromb. Hemost., 10(2), 109-121(1984) https://doi.org/10.1055/s-2007-1004413
  5. Joyce, D. E., Gelbert, L., Ciaccia, A., DeHoff, B. and Grinnell, B. W., 'Gene Expression Profile of Antithrombotic Protein c Defines New Mechanisms Modulating Inflammation and Apoptosis,' J. Biol. Chem., 264(14), 11199-11203(2001) https://doi.org/10.1074/jbc.C100017200
  6. Riewald, M., Petrovan, R. J., Donner, A., Mueller, B. M. and Ruf, W., "Activation of Endothelial Cell Protease Activated Receptor 1 by the Protein C Pathway," Science, 296(5574), 1880-1882(2002) https://doi.org/10.1126/science.1071699
  7. Mosnier, L. O. and Griffin, J. H., "Inhibition of Staurosporineinduced Apoptosis of Endothelial Cells by Activated Protein C Requires Protease-activated Receptor-1 And endothelial cell protein C receptor," Biochem. J., 373, 65-70(2003) https://doi.org/10.1042/BJ20030341
  8. Cheng, T., Liu, D., Griffin, J. H., Fernandez, J. A., Castellino, F., Rosen, E. D., Fukudome, K. and Zlokovic, B. V., "Activated Protein C Blocks p53-Mediated Apoptosis in Ischemic Human Brain Endothelium and is Neuroprotective," Nat. Med., 9, 338-342(2003) https://doi.org/10.1038/nm826
  9. Guo, H., Liu, D., Gelbard, H., Cheng, T., Insalaco, R., Fernandez, J. A., Griffin, J. H. and Zlokovic, B. V., "Activated Protein C Prevents Neuronal Apoptosis Via Protease Activated Receptors 1 and 3," Neuron, 41, 563-572(2004) https://doi.org/10.1016/S0896-6273(04)00019-4
  10. Bae, J. S., Yang, L., Manditody, C. and Rezaie, A. R., "The Ligand Occupancy of Endothelial Protein C Receptor Switches the Protease-activated Receptor 1-dependent Signaling Specific-ity of Thrombin from a Permeability-enhancing to a Barrier-protective Response in Endothelial Cells," Blood, 110(12), 3909-3916(2007) https://doi.org/10.1182/blood-2007-06-096651
  11. Bae, J. S., Yang, L., Manditody, C. and Rezaie, A. R., "Engineering a Disulfide Bond to Stabilize the Calcium-binding Loop of Activated Protein C Eliminates Its Anticoagulant But not its Protective Signaling Properties," J. Biol. Chem., 282(12), 9251-9259(2007) https://doi.org/10.1074/jbc.M610547200
  12. Bernard, G. R., Vincent, J. L., Laterre, P. F., LaRosa, S. P., Dhainaut, J. F., Lopez-Rodriguez, A., Steingrub, J. S., Garber, G. E., Helterbrand, J. D., Ely, E. W. and Fisher, C. J. Jr; Recombinant human protein C Worldwide Evaluation in Severe Sepsis(PROWESS) study group, "Efficacy and Safety of Recombinant Human Activated Protein C for Severe Sepsis," N. Engl. J. Med., 344, 699-709(2001) https://doi.org/10.1056/NEJM200103083441001
  13. Monnier, L. O., Zlokovic, B. V. and Griffin, J. H., "The Cytoprotective Protein C Pathway," Blood, 109(8), 3161-3172(2007) https://doi.org/10.1182/blood-2006-09-003004
  14. Esmon, S. T., 'The Protein C Pathway,' Chest, 124(3 supple), 26S-32S(2003) https://doi.org/10.1378/chest.124.3_suppl.26S
  15. Bae, J. S., Yang, L. and Rezaie, A. R., "Receptors of the Protein C Activation and Activated Protein C Signaling Pathways are Colocalized in Lipid Rafts of Endothelial Cells," Proc. Natl. Acad. Sci. U.S.A., 104(8), 2867-2872(2007) https://doi.org/10.1073/pnas.0611493104
  16. Fukudome, K., Kurosawa, S., Stearns-Kurosawa, D. J., He, X., Rezaie, A. R. and Esmon, C. T., "The Endothelial Cell Protein C Receptor. Cell Surface Expression and Direct Ligand Binding by the Soluble Receptor," J. Biol. Chem., 271, 17491-17498(1996) https://doi.org/10.1074/jbc.271.29.17491
  17. Regan, L. M., Mollica, J. S., Rezaie, A. R. and Esmon, C. T., "The Interaction Between the Endothelial Cell Protein C Receptor and Protein C Is Dictated by the gamma-Carboxyglutamic Acid Domain of Protein C," J. Biol. Chem., 272, 26279-26284(1997) https://doi.org/10.1074/jbc.272.42.26279
  18. Kurosawa, S., Stearns-Kurosawa, D. J., Hidari, D. and Esmon, C. T., "Identification of Functional Endothelial Protein C Receptor in Human Plasma," J. Clin. Invest., 100(2), 411-418(1997) https://doi.org/10.1172/JCI119548
  19. Kurosawa, S., Stearns-Kurosawa, D. J., Carson, C. W., D'Angelo, A., Della Valle, P. and Esmon, C. T., 'Plasma Levels of Endothelial Cell Protein C Receptor are Elevated in Patients with Sepsis and Systemic Lupus Erythematosus: Lack of Correlation with Thrombomodulin Suggests Involvement of Different Pathological Processes,' Blood, 91(2), 725-727(1998)
  20. Regan, L. M., Stearns-Kurosawa, D. J., Kurosawa, S., Mollica, J., Fukudome, K. and Esmon C. T., "The Endothelial Cell Protein C Receptor. Inhibition of Activated Protein C Anticoagulant Function Without Modulation of Reaction with Proteinase Inhibitors," J. Biol. Chem., 271(29), 17499-17503(1996) https://doi.org/10.1074/jbc.271.29.17499
  21. Bae, J. S. and Rezaie, A. R., 'Protease Activated Receptor 1(PAR-1) Activation by Thrombin is Protective in Human Pulmonary Artery Endothelial Cells If Endothelial Protein C Receptor is Occupied by Its Natural Ligand,' Semin, Thrombo. Haemost., 100(1), 101-109(2008)