Thromb Haemost 2019; 119(03): 467-478
DOI: 10.1055/s-0038-1677532
Stroke, Systemic or Venous Thromboembolism
Georg Thieme Verlag KG Stuttgart · New York

Long-Term Treatment with Thrombomodulin Improves Functional Outcomes after Cerebral Ischemia Even if Administration is Delayed

Takafumi Nakano
1   Department of Pharmaceutical and Health Care Management, Faculty of Pharmaceutical Sciences, Fukuoka University, Jyonan, Fukuoka, Japan
2   Department of Pharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, Jyonan, Fukuoka, Japan
3   Department of Pharmacy, Fukuoka University Hospital, Jonan, Fukuoka, Japan
,
Yoshihiko Nakamura
4   Department of Emergency and Critical Care Medicine, Fukuoka University Hospital, Jonan, Fukuoka, Japan
,
Kiyoshi Matsuyama
5   Department of Biochemistry and Applied Chemistry, Kurume National College of Technology, Kurume, Japan
,
Keiichi Irie
2   Department of Pharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, Jyonan, Fukuoka, Japan
,
Mako Yasumura
2   Department of Pharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, Jyonan, Fukuoka, Japan
,
Yurie Hirata
5   Department of Biochemistry and Applied Chemistry, Kurume National College of Technology, Kurume, Japan
,
Motoki Yamasaki
2   Department of Pharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, Jyonan, Fukuoka, Japan
,
Kanae Misumi
4   Department of Emergency and Critical Care Medicine, Fukuoka University Hospital, Jonan, Fukuoka, Japan
,
Yuta Yamashita
2   Department of Pharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, Jyonan, Fukuoka, Japan
,
Takayuki Myose
2   Department of Pharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, Jyonan, Fukuoka, Japan
,
Koichi Matsuo
1   Department of Pharmaceutical and Health Care Management, Faculty of Pharmaceutical Sciences, Fukuoka University, Jyonan, Fukuoka, Japan
,
Kazunori Sano
2   Department of Pharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, Jyonan, Fukuoka, Japan
,
Hidetoshi Kamimura
1   Department of Pharmaceutical and Health Care Management, Faculty of Pharmaceutical Sciences, Fukuoka University, Jyonan, Fukuoka, Japan
3   Department of Pharmacy, Fukuoka University Hospital, Jonan, Fukuoka, Japan
,
Hiroyasu Ishikura
4   Department of Emergency and Critical Care Medicine, Fukuoka University Hospital, Jonan, Fukuoka, Japan
,
Takashi Egawa
1   Department of Pharmaceutical and Health Care Management, Faculty of Pharmaceutical Sciences, Fukuoka University, Jyonan, Fukuoka, Japan
,
Kenichi Mishima
2   Department of Pharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, Jyonan, Fukuoka, Japan
› Author Affiliations
Funding None.
Further Information

Publication History

23 July 2018

09 December 2018

Publication Date:
21 January 2019 (online)

Abstract

Our previous study indicated that recombinant human soluble thrombomodulin (rhsTM) could attenuate brain damage when administered as a bolus in the cerebral ischaemic early phase. Then, we considered that treatment with rhsTM may show therapeutic effects even when administered in the ischaemic delayed phase, because rhsTM has an action of inhibiting high-mobility group box 1 (HMGB1) as a late mediator of lethal systemic inflammation. This study was performed to investigate the effects of delayed treatment with rhsTM on ischaemic brain damage induced by high HMGB1 level in mice subjected to 4-hour middle cerebral artery occlusion (MCAO). One day after MCAO, rhsTM was administered intraperitoneally at a dose of 1 or 5 mg/kg once a day for 7 days. Neurological score, motor coordination and HMGB1 levels were measured 1, 3 and 7 days after MCAO. The presence of activated microglia was evaluated 7 days after MCAO. Systemic HMGB1 levels increased 1 to 7 days after MCAO and were higher at 7 days compared with day 1. At the same time, survival rate decreased, and activated microglia increased in the infarct area. Treatment with rhsTM improved neurological score, motor coordination, survival and prevented brain damage. Moreover, rhsTM decreased both HMGB1 level and number of activated M1 microglia. The results of this study indicated that rhsTM improved functional outcomes via inhibition of HMGB1 up-regulation and M1 microglial activation in the cerebral ischaemic delayed phase. rhsTM may become a new therapeutic agent with a wide therapeutic time window in patients with cerebral ischaemia.

Authors' Contributions

T.N. performed the in vivo experiments, analysed the data and described the manuscript. Y.N., Mako Y., Motoki Y., Kanae M., Y.Y. and T.M. performed the in vivo experiments and analysed the data. Kioshi M. and Y.H. performed the in vitro experiments and analysed the data. K.I. interpreted the data and contributed most to the writing of the manuscript. Koichi M., K.S., H.K., H.I., T.E. and Kenichi M. interpreted the data and contributed to the writing of the manuscript. All authors read and approved the final manuscript.


Supplementary Material

 
  • References

  • 1 Hacke W, Kaste M, Fieschi C. , et al; The European Cooperative Acute Stroke Study (ECASS). Intravenous thrombolysis with recombinant tissue plasminogen activator for acute hemispheric stroke. JAMA 1995; 274 (13) 1017-1025
  • 2 Marler JR, Goldstein LB. Medicine. Stroke--tPA and the clinic. Science 2003; 301 (5640): 1677
  • 3 Saver JL, Levine SR. Alteplase for ischaemic stroke--much sooner is much better. Lancet 2010; 375 (9727): 1667-1668
  • 4 Ikezoe T. Thrombomodulin/activated protein C system in septic disseminated intravascular coagulation. J Intensive Care 2015; 3 (01) 1
  • 5 Mohri M, Sugimoto E, Sata M, Asano T. The inhibitory effect of recombinant human soluble thrombomodulin on initiation and extension of coagulation--a comparison with other anticoagulants. Thromb Haemost 1999; 82 (06) 1687-1693
  • 6 Olivot JM, Labreuche J, Aiach M, Amarenco P. ; GENIC Investigators. Soluble thrombomodulin and brain infarction: case-control and prospective study. Stroke 2004; 35 (08) 1946-1951
  • 7 Aikawa N, Shimazaki S, Yamamoto Y. , et al. Thrombomodulin alfa in the treatment of infectious patients complicated by disseminated intravascular coagulation: subanalysis from the phase 3 trial. Shock 2011; 35 (04) 349-354
  • 8 Yamakawa K, Fujimi S, Mohri T. , et al. Treatment effects of recombinant human soluble thrombomodulin in patients with severe sepsis: a historical control study. Crit Care 2011; 15 (03) R123
  • 9 Saito H, Maruyama I, Shimazaki S. , et al. Efficacy and safety of recombinant human soluble thrombomodulin (ART-123) in disseminated intravascular coagulation: results of a phase III, randomized, double-blind clinical trial. J Thromb Haemost 2007; 5 (01) 31-41
  • 10 Moll S, Lindley C, Pescatore S. , et al. Phase I study of a novel recombinant human soluble thrombomodulin, ART-123. J Thromb Haemost 2004; 2 (10) 1745-1751
  • 11 Kearon C, Comp P, Douketis J, Royds R, Yamada K, Gent M. Dose-response study of recombinant human soluble thrombomodulin (ART-123) in the prevention of venous thromboembolism after total hip replacement. J Thromb Haemost 2005; 3 (05) 962-968
  • 12 Ito T, Kawahara K, Okamoto K. , et al. Proteolytic cleavage of high mobility group box 1 protein by thrombin-thrombomodulin complexes. Arterioscler Thromb Vasc Biol 2008; 28 (10) 1825-1830
  • 13 Yamato M, Minematsu Y, Fujii J. , et al. Effective combination therapy of polymyxin-B direct hemoperfusion and recombinant thrombomodulin for septic shock accompanied by disseminated intravascular coagulation: a historical controlled trial. Ther Apher Dial 2013; 17 (05) 472-476
  • 14 Ruf W, Edgington TS. Structural biology of tissue factor, the initiator of thrombogenesis in vivo. FASEB J 1994; 8 (06) 385-390
  • 15 Levi M, Keller TT, van Gorp E, ten Cate H. Infection and inflammation and the coagulation system. Cardiovasc Res 2003; 60 (01) 26-39
  • 16 Zeerleder S, Hack CE, Wuillemin WA. Disseminated intravascular coagulation in sepsis. Chest 2005; 128 (04) 2864-2875
  • 17 Semeraro N, Ammollo CT, Semeraro F, Colucci M. Sepsis, thrombosis and organ dysfunction. Thromb Res 2012; 129 (03) 290-295
  • 18 Harris HE, Raucci A. Alarmin(g) news about danger: workshop on innate danger signals and HMGB1. EMBO Rep 2006; 7 (08) 774-778
  • 19 Allam R, Kumar SV, Darisipudi MN, Anders HJ. Extracellular histones in tissue injury and inflammation. J Mol Med (Berl) 2014; 92 (05) 465-472
  • 20 Zhang Q, Raoof M, Chen Y. , et al. Circulating mitochondrial DAMPs cause inflammatory responses to injury. Nature 2010; 464 (7285): 104-107
  • 21 Wang H, Bloom O, Zhang M. , et al. HMG-1 as a late mediator of endotoxin lethality in mice. Science 1999; 285 (5425): 248-251
  • 22 Wang H, Yang H, Czura CJ, Sama AE, Tracey KJ. HMGB1 as a late mediator of lethal systemic inflammation. Am J Respir Crit Care Med 2001; 164 (10 Pt 1): 1768-1773
  • 23 Kim JB, Sig Choi J, Yu YM. , et al. HMGB1, a novel cytokine-like mediator linking acute neuronal death and delayed neuroinflammation in the postischemic brain. J Neurosci 2006; 26 (24) 6413-6421
  • 24 Hayakawa K, Mishima K, Nozako M. , et al. Delayed treatment with minocycline ameliorates neurologic impairment through activated microglia expressing a high-mobility group box1-inhibiting mechanism. Stroke 2008; 39 (03) 951-958
  • 25 Nakamura Y, Nakano T, Irie K. , et al. Recombinant human soluble thrombomodulin ameliorates cerebral ischemic injury through a high-mobility group box 1 inhibitory mechanism without hemorrhagic complications in mice. J Neurol Sci 2016; 362: 278-282
  • 26 Hayakawa K, Mishima K, Irie K. , et al. Cannabidiol prevents a post-ischemic injury progressively induced by cerebral ischemia via a high-mobility group box1-inhibiting mechanism. Neuropharmacology 2008; 55 (08) 1280-1286
  • 27 Yang Y, Liu H, Zhang H. , et al. ST2/IL-33-dependent microglial response limits acute ischemic brain injury. J Neurosci 2017; 37 (18) 4692-4704
  • 28 Sauerbrey F. Verwendung von schwingquarzen zur wägung dünner schichten und microwägung. Z Phys 1959; 155 (02) 206-222
  • 29 Okahata Y, Niikura K, Sugiura Y, Sawada M, Morii T. Kinetic studies of sequence-specific binding of GCN4-bZIP peptides to DNA strands immobilized on a 27-MHz quartz-crystal microbalance. Biochemistry 1998; 37 (16) 5666-5672
  • 30 Gu JJ, Chen JB, Zhang JH, Zhang H, Wang SS. Recombinant human soluble thrombomodulin protects against brain injury in a CVST rat model, via downregulation of the HMGB1-RAGE axis. Mol Med Rep 2016; 14 (06) 5217-5222
  • 31 Ryang YM, Dang J, Kipp M. , et al. Solulin reduces infarct volume and regulates gene-expression in transient middle cerebral artery occlusion in rats. BMC Neurosci 2011; 12: 113
  • 32 Su EJ, Geyer M, Wahl M. , et al. The thrombomodulin analog Solulin promotes reperfusion and reduces infarct volume in a thrombotic stroke model. J Thromb Haemost 2011; 9 (06) 1174-1182
  • 33 Haruma J, Teshigawara K, Hishikawa T. , et al. Anti-high mobility group box-1 (HMGB1) antibody attenuates delayed cerebral vasospasm and brain injury after subarachnoid hemorrhage in rats. Sci Rep 2016; 6: 37755
  • 34 Hori O, Brett J, Slattery T. , et al. The receptor for advanced glycation end products (RAGE) is a cellular binding site for amphoterin. Mediation of neurite outgrowth and co-expression of rage and amphoterin in the developing nervous system. J Biol Chem 1995; 270 (43) 25752-25761
  • 35 Das N, Dewan V, Grace PM. , et al. HMGB1 activates proinflammatory signaling via TLR5 leading to allodynia. Cell Reports 2016; 17 (04) 1128-1140
  • 36 Brett J, Schmidt AM, Yan SD. , et al. Survey of the distribution of a newly characterized receptor for advanced glycation end products in tissues. Am J Pathol 1993; 143 (06) 1699-1712
  • 37 Thornalley PJ. Cell activation by glycated proteins. AGE receptors, receptor recognition factors and functional classification of AGEs. Cell Mol Biol 1998; 44 (07) 1013-1023
  • 38 Sousa MM, Yan SD, Stern D, Saraiva MJ. Interaction of the receptor for advanced glycation end products (RAGE) with transthyretin triggers nuclear transcription factor kB (NF-kB) activation. Lab Invest 2000; 80 (07) 1101-1110
  • 39 Morioka T, Kalehua AN, Streit WJ. The microglial reaction in the rat dorsal hippocampus following transient forebrain ischemia. J Cereb Blood Flow Metab 1991; 11 (06) 966-973
  • 40 Lemstra AW, Groen in't Woud JC, Hoozemans JJ. , et al. Microglia activation in sepsis: a case-control study. J Neuroinflammation 2007; 4: 4
  • 41 Takizawa T, Shibata M, Kayama Y. , et al. High-mobility group box 1 is an important mediator of microglial activation induced by cortical spreading depression. J Cereb Blood Flow Metab 2017; 37 (03) 890-901
  • 42 Pan J, Jin JL, Ge HM. , et al. Malibatol A regulates microglia M1/M2 polarization in experimental stroke in a PPARγ-dependent manner. J Neuroinflammation 2015; 12: 51
  • 43 Higashi Y, Aratake T, Shimizu S. , et al. Influence of extracellular zinc on M1 microglial activation. Sci Rep 2017; 7: 43778
  • 44 Qin C, Fan WH, Liu Q. , et al. Fingolimod protects against ischemic white matter damage by modulating microglia toward M2 polarization via STAT3 pathway. Stroke 2017; 48 (12) 3336-3346
  • 45 Xiong XY, Liu L, Yang QW. Functions and mechanisms of microglia/macrophages in neuroinflammation and neurogenesis after stroke. Prog Neurobiol 2016; 142: 23-44
  • 46 Chen S, Dong Z, Cheng M. , et al. Homocysteine exaggerates microglia activation and neuroinflammation through microglia localized STAT3 overactivation following ischemic stroke. J Neuroinflammation 2017; 14 (01) 187
  • 47 Huang M, Wan Y, Mao L. , et al. Inhibiting the migration of M1 microglia at hyperacute period could improve outcome of tMCAO rats. CNS Neurosci Ther 2017; 23 (03) 222-232
  • 48 Wolter J, Schild L, Bock F. , et al. Thrombomodulin-dependent protein C activation is required for mitochondrial function and myelination in the central nervous system. J Thromb Haemost 2016; 14 (11) 2212-2226
  • 49 Pathak R, Wang J, Garg S, Aykin-Burns N, Petersen KU, Hauer-Jensen M. Recombinant thrombomodulin (Solulin) ameliorates early intestinal radiation toxicity in a preclinical rat model. Radiat Res 2016; 186 (02) 112-120
  • 50 Aoki Y, Takei R, Mohri M. , et al. Antithrombotic effects of recombinant human soluble thrombomodulin (rhs-TM) on arteriovenous shunt thrombosis in rats. Am J Hematol 1994; 47 (03) 162-166