Focal arterial inflammation is augmented in mice with a deficiency of the protein C gene

Summary

Increased risk of thrombosis, with propitious conditions for fibrin deposition, along with upregulation of inflammation, are important factors that enhance plaque formation in atherosclerosis. Evidence supporting the role of anticoagulant protein C (PC) as an inflammatory agent has emerged, supplementing its wellknown function as an anticoagulant. Thus, we sought to examine whethera PC deficiency would lead to an enhanced response to an acute arterial hyperplasic challenge. The presentation of early arterial inflammation was studied using a copper/silicone arterial cuff model of accelerated focal neointimal remodeling in mice with a heterozygous total deficiency of PC (PC^+/−). Increased inflammation, cell proliferation, cell migration, fibrin elevation, and tissue necrosis were observed in the treated arteries of PC^+/−mice, as compared to arteries of equally challenged age- and gender-matched WT mice. These results indicate that PC^+/−mice subjected to this challenge displayed enhanced focal arterial inflammation and thrombosis, leading to larger neointimas and subsequent localized occlusion, as compared to their WT counterparts.

Keywords

Inflammation - thrombosis - arterial injury - arterial remodeling - arterial neointima formation

Footnote:

J.G.G., M.J.S-C., and F.N. contributed equally to the performance of the experiments.

• References

• 1 Griffin JH, Evatt B, Wideman C. et al. Anticoagulant proteinC pathway defective in majority of thrombophilic patients. Blood 1993; 83: 2008-9.
• 2 Castellino FJ. Gene targeting in hemostasis: Protein C. Front Biosci 2001; 06: D807-D19.
• 3 Joyce DE, Gelbert L, Ciaccia A. et al. Gene expression profile of antithrombotic protein C defines new mechanisms modulating inflammation and apoptosis. J Biol Chem 2001; 276: 11199-203.
• 4 Kirschenbaum LA, McKevitt D, Rullan M. et al. Importance of platelets and fibrinogen in neutrophilendothelial cell interactions in septic shock. Crit Care Med 2004; 32: 1094-9.
• 5 O’Brien LA, Gupta A, Grinnell BW. Activated protein C and sepsis. Front Biosci 2006; 11: 676-98.
• 6 Cheng T, Liu D, Griffin JH. et al. Activated Protein C blocks p53-mediated apoptosis in ischemic human brain endothelium and protects in a murine stroke model through EPCR and PAR-1. Nature Med 2003; 09: 338-42.
• 7 Guo H, Liu D, Gelbard H. et al. Activated proteinC prevents neuronal apoptosis via protease activated receptors 1 and 3. Neuron 2004; 41: 563-72.
• 8 Liu D, Cheng T, Guo H. et al. Tissue plasminogen activator neurovascular toxicity is controlled by activated protein C. Nat Med 2004; 10: 1379-83.
• 9 Jalbert LR, Rosen ED, Moons L. et al. Inactivation of the gene for anticoagulant Protein C causes lethal perinatal consumptive coagulopathy in mice. J Clin Invest 1998; 102: 1481-8.
• 10 Lay AJ, Zhong L, Rosen ED. et al. Mice witha severe deficiency of proteinC display prothrombotic and proinflammatory phenotypes and compromised maternal reproductive capabilities. J Clin Invest 2005; 115: 1552-6.
• 11 Murakami K, Okajima K, Uchiba M. et al. Activated proteinC attenuates endotoxin-induced pulmonary vascular injury by inhibiting activated leukocytes in rats. Blood 1996; 87: 642-7.
• 12 Shibata M, Kumar SR, Amar A. et al. Anti-inflammatory, antithrombotic, and neuroprotective effects of activated proteinC in a murine model of focal ischemic stroke. Circulation 2001; 103: 1799-805.
• 13 Bernard GR, Vincent JL, Laterre PF. et al. Efficacy and safety of recombinant human activated protein C for severe sepsis. N Engl J Med 2001; 344: 699-709.
• 14 Dhainaut JF, Yan SB, Margolis BD. et al. Drotrecogin alfa (activated) (recombinant human activated protein C) reduces host coagulopathy response in patients with severe sepsis. Thromb Haemost 2003; 90: 642-53.
• 15 Tsuchida A, Thomson NM, Salem HH. et al. Serial monitoring shows plasma proteinC and free proteinS levels are decreased during human acute renal allograft rejection. Transplant Proc 1990; 22: 2134-6.
• 16 Kiechl S, Muigg A, Santer P. et al. Poor response to activated proteinC as a prominent risk predictor of advanced atherosclerosis and arterial disease. Circulation 1999; 99: 614-9.
• 17 Kario K, Sakata T, Miyata T. et al. Is acquired activated proteinC resistance a cardiovascular risk?. Circulation 2000; 101: E120.
• 18 Ploplis VA, Cornelissen I, Sandoval-Cooper MJ. et al. Remodeling of the vessel wall after copper-induced injury is highly attenuated in mice with a total deficiency of plasminogen activator inhibitor-1. Am J Pathol 2001; 158: 107-17.
• 19 Reunanen A, Knekt P, Marniemi J. et al. Serum calcium, magnesium, copper and zinc and risk of cardiovascular death. Eur J Clin Nutr 1996; 50: 431-7.
• 20 Mansoor MA, Bergmark C, Haswell SJ. et al. Correlation between plasma total homocysteine and copper in patients with peripheral vascular disease. Clin Chem 2000; 46: 385-91.
• 21 Ganopolsky JG, Castellino FJ. A protein C deficiency exacerbates inflammatory and hypotensive responses in mice during polymicrobial sepsis in a cecal ligation and puncture model. Am J Pathol 2004; 165: 1433-46.
• 22 Masson P. Trichrome stainings and their preliminary technique. J Tech Meth 1929; 12: 75-90.
• 23 Sheehan D, Hrapchak B. Connective Tissue and Muscle Fiber Stains. In: Theory and Practice of Histotechnology. Battelle Press; Columbus, Ohio, USA: 1987: 196.
• 24 Sato J, Schorey J, Ploplis VA. et al. The fibrinolytic system in dissemination and matrix protein deposition duringa Mycobacterium infection. Am J Pathol 2003; 163: 517-31.
• 25 Volker W, Dorszewski A, Unruh V. et al. Copper-induced inflammatory reactions of rat carotid arteries mimic restenosis/arteriosclerosis-like neointima formation. Atherosclerosis 1997; 130: 29-36.
• 26 Ross R. Atherosclerosis--an inflammatory disease. N Engl J Med 1999; 340: 115-26.
• 27 Gerszten RE, Garcia-Zepeda EA, Lim YC. et al. MCP-1 and IL-8 trigger firm adhesion of monocytes to vascular endothelium under flow conditions. Nature 1999; 398: 718-23.
• 28 Feistritzer C, Riewald M. Endothelial barrier protection by activated proteinC through PAR1-dependent sphingosine 1-phosphate receptor-1 crossactivation. Blood 2005; 105: 3178-84.
• 29 Finigan JH, Dudek SM, Singleton PA. et al. Activated proteinC mediates novel lung endothelial barrier enhancement: role of sphingosine 1-phosphate receptor transactivation. J Biol Chem 2005; 280: 17286-93.
• 30 Sturn DH, Kaneider NC, Feistritzer C. et al. Expression and function of the endothelial protein C receptor in human neutrophils. Blood 2003; 102: 1499-505.
• 31 Grey ST, Tsuchida A, Hau H. et al. Selective inhibitory effects of the anticoagulant activated proteinC on the responses of human mononuclear phagocytes to LPS, IFN-g, or phorbol ester. J Immunol 1994; 153: 3664-72.
• 32 Hancock WW, Grey ST, Hau L. et al. Binding of activated proteinC to a specific receptor on human mononuclear phagocytes inhibits intracellular calcium signaling and monocyte-dependent proliferative responses. Transplantation 1995; 60: 1525-32.
• 33 Grey ST, Hancock WW. A physiologic anti-inflammatory pathway based on thrombomodulin expression and generation of activated proteinC by human mononuclear phagocytes. J Immunol 1996; 156: 2256-63.
• 34 Uchiba M, Okajima K, Oike Y. et al. Activated protein C induces endothelial cell proliferation by mitogen-activated protein kinase activation in vitro and angiogenesis in vivo . Circ Res 2004; 95: 34-41.