Urokinase-type plasminogen activator contributes to heterogeneity of macrophages at the border of damaged site during liver repair in mice

Naoyuki Kawao; Nobuo Nagai; Yukinori Tamura; Kiyotaka Okada; Masato Yano; Yasuhiro Suzuki; Kazuo Umemura; Shigeru Ueshima; Osamu Matsuo

doi:10.1160/TH10-08-0516

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2011; 105(05): 892-900
DOI: 10.1160/TH10-08-0516

Wound Healing and Inflammation/Infection

Schattauer GmbH

Urokinase-type plasminogen activator contributes to heterogeneity of macrophages at the border of damaged site during liver repair in mice

Naoyuki Kawao

¹Department of Physiology, Kinki University Faculty of Medicine, Osakasayama, Japan

,

Nobuo Nagai

¹Department of Physiology, Kinki University Faculty of Medicine, Osakasayama, Japan

²Department of Animal Bio-science, Faculty of Bio-science, Nagahama Institute of Bio-Science and Technology, Nagahama, Japan

,

Yukinori Tamura

¹Department of Physiology, Kinki University Faculty of Medicine, Osakasayama, Japan

,

Kiyotaka Okada

¹Department of Physiology, Kinki University Faculty of Medicine, Osakasayama, Japan

,

Masato Yano

¹Department of Physiology, Kinki University Faculty of Medicine, Osakasayama, Japan

,

Yasuhiro Suzuki

³Department of Pharmacology, Hamamatsu University School of Medicine, Hamamatsu, Japan

,

Kazuo Umemura

³Department of Pharmacology, Hamamatsu University School of Medicine, Hamamatsu, Japan

,

Shigeru Ueshima

¹Department of Physiology, Kinki University Faculty of Medicine, Osakasayama, Japan

⁴Department of Food Science and Nutrition, Kinki University School of Agriculture, Nara, Japan

,

Osamu Matsuo

¹Department of Physiology, Kinki University Faculty of Medicine, Osakasayama, Japan

› Author Affiliations

Abstract

Summary

Urokinase-type plasminogen activator (u-PA) plays an important role in tissue remodelling through the activation of plasminogen in the liver, but its mechanisms are less well known. Here, we investigated the involvement of u-PA in the accumulation and phenotypic heterogeneity of macrophages at the damaged site during liver repair. After induction of liver injury by photochemical reaction in mice, the subsequent pathological responses and expression of phenotypic markers in activated macrophages were analysed histologically. Fibrinolytic activity at the damaged site was also examined by fibrin zymography. In wild-type mice, the extent of damage decreased gradually until day 14 and was associated with an accumulation of macrophages at the border of the damaged site. In addition, the macrophages that accumulated near the damaged tissue expressed CD206, a marker of highly phagocytic macrophages, on day 7. Further, macrophages that were adjacent to CD206-positive cells expressed inducible nitric oxide synthase (iNOS), a pro-inflammatory marker. u-PA activity increased at the damaged site on days 4 and 7, which distributed primarily at the border region. In contrast, in u-PA-deficient mice, the decrease in damage size and the accumulation of macrophages were impaired. Further, neither CD206 nor iNOS was expressed in the macrophages that accumulated at the border region in u-PA-deficient mice. Mice deficient for the gene encoding either u-PA receptor (u-PAR) or tissue-type plasminogen activator experienced normal recovery during liver repair. These data indicate that u-PA mediates the accumulation of macrophages and their phenotypic heterogeneity at the border of damaged sites through u-PAR-independent mechanisms.

Keywords

u-PA - macrophage accumulation - macrophage activation - macrophage heterogeneity - liver repair

Full Text

References

References
1 Singer AJ, Clark RA. Cutaneous wound healing. N Engl J Med 1999; 341: 738-746.
2 Mirabelli-Badenier M, Braunersreuther V, Viviani GL. et al. CC and CXC chemokines are pivotal mediators of cerebral injury in ischaemic stroke. Thromb Haemost. 2011 105: prepublished online.
3 Soehnlein O, Zernecke A, Weber C. Neutrophils launch monocyte extravasation by release of granule proteins. Thromb Haemost 2009; 102: 198-205.
4 Duffield JS, Forbes SJ, Constandinou CM. et al. Selective depletion of macrophages reveals distinct, opposing roles during liver injury and repair. J Clin Invest 2005; 115: 56-65.
5 Kawao N, Nagai N, Ishida C. et al. Plasminogen is essential for granulation tissue formation during the recovery process after liver injury in mice. J Thromb Haemost 2010; 8: 1555-1566.
6 Stout RD, Suttles J. Functional plasticity of macrophages: reversible adaptation to changing microenvironments. J Leukoc Biol 2004; 76: 509-513.
7 Newby AC, George SJ, Ismail Y. et al. Vulnerable atherosclerotic plaque metalloproteinases and foam cell phenotypes. Thromb Haemost 2009; 101: 1006-1011.
8 Mosser DM, Edwards JP. Exploring the full spectrum of macrophage activation. Nat Rev Immunol 2008; 8: 958-969.
9 Gazi U, Martinez-Pomares L. Influence of the mannose receptor in host immune responses. Immunobiology 2009; 214: 554-561.
10 Aderem A, Underhill DM. Mechanisms of phagocytosis in macrophages. Annu Rev Immunol 1999; 17: 593-623.
11 Castellino FJ, Ploplis VA. Structure and function of the plasminogen/plasmin system. Thromb Haemost 2005; 93: 647-654.
12 Blasi F, Carmeliet P. uPAR: a versatile signalling orchestrator. Nat Rev Mol Cell Biol 2002; 3: 932-943.
13 Nagai N, Okada K, Kawao N. et al. Urokinase-type plasminogen activator receptor (uPAR) augments brain damage in a murine model of ischemic stroke. Neurosci Lett 2008; 432: 46-49.
14 Nagai N, De Mol M, Lijnen HR. et al. Role of plasminogen system components in focal cerebral ischemic infarction: a gene targeting and gene transfer study in mice. Circulation 1999; 99: 2440-2444.
15 Bezerra JA, Bugge TH, Melin-Aldana H. et al. Plasminogen deficiency leads to impaired remodeling after a toxic injury to the liver. Proc Natl Acad Sci USA 1999; 96: 15143-15148.
16 Bezerra JA, Currier AR, Melin-Aldana H. et al. Plasminogen activators direct reorganization of the liver lobule after acute injury. Am J Pathol 2001; 158: 921-929.
17 Ng VL, Sabla GE, Melin-Aldana H. et al. Plasminogen deficiency results in poor clearance of non-fibrin matrix and persistent activation of hepatic stellate cells after an acute injury. J Hepatol 2001; 35: 781-789.
18 Okada K, Ueshima S, Imano M. et al. The regulation of liver regeneration by the plasmin/alpha 2-antiplasmin system. J Hepatol 2004; 40: 110-116.
19 Kawao N, Okada K, Kawata S. et al. Plasmin decreases the BH3-only protein BimEL via the ERK1/2 signaling pathway in hepatocytes. Biochim Biophys Acta 2007; 1773: 718-727.
20 Okada K, Ueshima S, Kawao N. et al. Binding of plasminogen to hepatocytes isolated from injured mice liver and nonparenchymal cell-dependent proliferation of hepatocytes. Blood Coagul Fibrinolysis 2008; 19: 503-511.
21 Shimizu M, Hara A, Okuno M. et al. Mechanism of retarded liver regeneration in plasminogen activator-deficient mice: impaired activation of hepatocyte growth factor after Fas-mediated massive hepatic apoptosis. Hepatology 2001; 33: 569-576.
22 Porcheray F, Viaud S, Rimaniol AC. et al. Macrophage activation switching: an asset for the resolution of inflammation. Clin Exp Immunol 2005; 142: 481-489.
23 Kawao N, Nagai N, Okada K. et al. Role of plasminogen in macrophage accumulation during liver repair. Thromb Res 2010; 125: e214-221.
24 Kamanaka Y, Kawabata A, Matsuya H. et al. Effect of a potent iNOS inhibitor (ONO-1714) on acetaminophen-induced hepatotoxicity in the rat. Life Sci 2003; 74: 793-802.
25 Nagai N, Kawao N, Okada K. et al. Initial brain lesion size affects the extent of subsequent pathophysiological responses. Brain Res 2010; 1322: 109-117.
26 Nagai N, Kawao N, Okada K. et al. Systemic transplantation of embryonic stem cells accelerates brain lesion decrease and angiogenesis. Neuroreport 2010; 21: 575-579.
27 Umemura K, Kohno Y, Matsuno H. et al. A new model for photochemically induced thrombosis in the inner ear microcirculation and the use of hearing loss as a measure for microcirculatory disorders. Eur Arch Otorhinolaryngol 1990; 248: 105-108.
28 Suzuki Y, Nagai N, Umemura K. et al. Stromelysin-1 (MMP-3) is critical for intracranial bleeding after t-PA treatment of stroke in mice. J Thromb Haemost 2007; 5: 1732-1739.
29 Suzuki Y, Nagai N, Yamakawa K. et al. Tissue-type plasminogen activator (t-PA) induces stromelysin-1 (MMP-3) in endothelial cells through activation of lipo-protein receptor-related protein. Blood 2009; 114: 3352-3358.
30 Carmeliet P, Schoonjans L, Kieckens L. et al. Physiological consequences of loss of plasminogen activator gene function in mice. Nature 1994; 368: 419-424.
31 Dewerchin M, Nuffelen AV, Wallays G. et al. Generation and characterization of urokinase receptor-deficient mice. J Clin Invest 1996; 97: 870-878.
32 Prasad S, McDaid JP, Tam FW. et al. Pkd2 dosage influences cellular repair responses following ischemia-reperfusion injury. Am J Pathol 2009; 175: 1493-1503.
33 Hou Y, Okamoto C, Okada K. et al. c-Myc is essential for urokinase plasminogen activator expression on hypoxia-induced vascular smooth muscle cells. Cardiovasc Res 2007; 75: 186-194.
34 Sheehan JJ, Zhou C, Gravanis I. et al. Proteolytic activation of monocyte chemoattractant protein-1 by plasmin underlies excitotoxic neurodegeneration in mice. J Neurosci 2007; 27: 1738-1745.
35 Gong Y, Hart E, Shchurin A. et al. Inflammatory macrophage migration requires MMP-9 activation by plasminogen in mice. J Clin Invest 2008; 118: 3012-3024.
36 Gordon S. Alternative activation of macrophages. Nat Rev Immunol 2003; 3: 23-35.
37 Stout RD, Jiang C, Matta B. et al. Macrophages sequentially change their functional phenotype in response to changes in microenvironmental influences. J Immunol 2005; 175: 342-349.
38 Bryer SC, Fantuzzi G, Van Rooijen N. et al. Urokinase-type plasminogen activator plays essential roles in macrophage chemotaxis and skeletal muscle regeneration. J Immunol 2008; 180: 1179-1188.
39 Kwon M, MacLeod TJ, Zhang Y. et al. S100A10, annexin A2, and annexin a2 heterotetramer as candidate plasminogen receptors. Front Biosci 2005; 10: 300-325.
40 Das R, Burke T, Plow EF. Histone H2B as a functionally important plasminogen receptor on macrophages. Blood 2007; 110: 3763-3772.
41 Li Q, Laumonnier Y, Syrovets T. et al. Plasmin triggers cytokine induction in human monocyte-derived macrophages. Arterioscler Thromb Vasc Biol 2007; 27: 1383-1389.
42 Tarui T, Akakura N, Majumdar M. et al. Direct interaction of the kringle domain of urokinase-type plasminogen activator (uPA) and integrin alpha v beta 3 induces signal transduction and enhances plasminogen activation. Thromb Haemost 2006; 95: 524-534.