Download PDF
Thromb Haemost 2006; 95(01): 166-173
DOI: 10.1160/TH05-07-0477
Animal Models
Schattauer GmbH

Treatment of embolic stroke in rats with bortezomib and recombinant human tissue plasminogen activator

Li Zhang
1   Department of Neurology
,
Zheng Gang Zhang
1   Department of Neurology
,
Xianshuang Liu
1   Department of Neurology
,
Ann Hozeska
1   Department of Neurology
,
Nancy Stagliano
2   Millennium Pharmaceuticals, Inc., Cambridge, Massachusetts, USA
,
William Riordan
2   Millennium Pharmaceuticals, Inc., Cambridge, Massachusetts, USA
,
Mei Lu
3   Department of Biostatistics and Research Epidemiology Henry Ford Health Sciences Center, Detroit, Michigan, USA
,
Michael Chopp
1   Department of Neurology
4   Department of Physics, Oakland University, Rochester, Michigan, USA
› Author AffiliationsThis work was supported by NINDS grants PO1 NS23393, RO1 NS38292, and RO1 HL64766.
Further Information

Publication History

Received 07 July 2005

Accepted after resubmission 07 November 2005

Publication Date:
28 November 2017 (online)

Also available at
    Permissions and Reprints

Summary

Stroke elicits a progressive vascular dysfunction, which contributes to the evolution of brain injury. Thrombolysis with tissue plasminogen activator (tPA) promotes adverse vascular events that limit the therapeutic window of stroke to three hours. Proteasome inhibitors reduce vascular thrombotic and inflammatory events, and consequently protect vascular function. The present study evaluated the neuroprotective effect of bortezomib,a potent and selective inhibitor of the proteasome, alone and in combination with delayed thrombolytic therapy on a rat model of embolic focal cerebral ischemia. Treatment with bortezomib reduces adverse cerebrovascular events including secondary thrombosis,inflammatory responses,and blood brain barrier (BBB) disruption, and hence reduces infarct volume and neurological functional deficit when administrated within 4 h after stroke onset. Combination of bortezomib and tPA extends the thrombolytic window for stroke to6 h, which is associated with the improvement of vascular patency and integrity. Real time RT-PCR of endothelial cells isolated by laser-capture microdissection from brain tissue and Western blot analysis showed that bortezomib upregulates endothelial nitric oxide synthase (eNOS) expression and blocks NF-κB activation. These results demonstrate that bortezomib promotes eNOS dependent vascular protection, and reduces NF-κB dependent vascular disruption, all of which may contribute to neuroprotection after stroke.

Keywords

Animal models - adhesion molecules - plasminogen activators
 

  • References

  • 1 van Gijn J, Dennis MS. Issues and answers in stroke care. Lancet 1998; 352 (Suppl. 03) SIII23-7.
    PubMedGoogle Scholar
  • 2 del Zoppo G, Ginis I, Hallenbeck JM. et al. Inflammation and stroke: putative role for cytokines, adhesion molecules and iNOS in brain response to ischemia. Brain Pathol 2000; 10: 95-112.
    PubMedGoogle Scholar
  • 3 Fisher M, Francis R. Altered coagulation in cerebral ischemia. Platelet, thrombin, and plasmin activity. Arch Neurol 1990; 47: 1075-9.
    CrossrefPubMedGoogle Scholar
  • 4 Zhang ZG, Chopp M, Goussev A. et al. Cerebral microvascular obstruction by fibrin is associated with upregulation of PAI-1 acutely after onset of focal embolic ischemia in rats. J Neurosci 1999; 19: 10898-907.
    CrossrefPubMedGoogle Scholar
  • 5 Chopp M, Zhang ZG. Anti-adhesion molecule and nitric oxide protection strategies in ischemic stroke. Curr Opin Neurol 1996; 09: 68-72.
    CrossrefPubMedGoogle Scholar
  • 6 Jean WC, Spellman SR, Nussbaum ES. et al. Reperfusion injury after focal cerebral ischemia: the role of inflammation and the therapeutic horizon. Neurosurgery 1998; 43: 1382-96.
    PubMedGoogle Scholar
  • 7 Coux O, Tanaka K, Goldberg AL. Structure and functions of the 20S and 26S proteasomes. Annu Rev Biochem 1996; 65: 801-47.
    CrossrefPubMedGoogle Scholar
  • 8 Goldberg AL. Functions of the proteasome: the lysis at the end of the tunnel. Science 1995; 268: 522-3.
    CrossrefPubMedGoogle Scholar
  • 9 Di Napoli M, Papa F. MLN-519. Millennium/ PAION. Curr Opin Investig Drugs 2003; 04: 333-41.
    PubMedGoogle Scholar
  • 10 Pye J, Ardeshirpour F, McCain A. et al. Proteasome inhibition ablates activation of NF-kappa B in myocardial reperfusion and reduces reperfusion injury. Am J Physiol Heart Circ Physiol 2003; 284: H919-26.
    CrossrefPubMedGoogle Scholar
  • 11 Stangl V, Lorenz M, Meiners S. et al. Long-term upregulation of eNOS and improvement of endothelial function by inhibition of the ubiquitin-proteasome pathway. Faseb J 2004; 18: 272-9.
    CrossrefPubMedGoogle Scholar
  • 12 Williams AJ, Hale SL, Moffett JR. et al. Delayed treatment with MLN519 reduces infarction and associated neurologic deficit caused by focal ischemic brain injury in rats via antiinflammatory mechanisms involving nuclear factor-kappaB activation, gliosis, and leukocyte infiltration. J Cereb Blood Flow Metab 2003; 23: 75-87.
    CrossrefPubMedGoogle Scholar
  • 13 Zhang RL, Zhang ZG, Chopp M. et al. Thrombolysis with tissue plasminogen activator alters adhesion molecule expression in the ischemic rat brain. Stroke 1999; 30: 624-9.
    CrossrefPubMedGoogle Scholar
  • 14 Sumii T, Lo EH. Involvement of matrix metalloproteinase in thrombolysis-associated hemorrhagic transformation after embolic focal ischemia in rats. Stroke 2002; 33: 831-6.
    CrossrefPubMedGoogle Scholar
  • 15 Zhang L, Zhang ZG, Zhang RL. et al. Postischemic (6-hour) treatment with recombinant human tissue plasminogen activator and proteasome inhibitor PS-519 reduces infarction in a rat model of embolic focal cerebral ischemia. Stroke 2001; 32: 2926-31.
    CrossrefPubMedGoogle Scholar
  • 16 Zivin J, Mazzarella V. Tissue plasminogen activator plus glutamate antagonist improves outcome after embolic stroke. Arch Neurol 1991; 48: 1235-8.
    CrossrefPubMedGoogle Scholar
  • 17 Palombella VJ, Conner EM, Fuseler JW. et al. Role of the proteasome and NF-kappaB in streptococcal cell wall-induced polyarthritis. Proc Natl Acad Sci U S A 1998; 95: 15671-6.
    CrossrefPubMedGoogle Scholar
  • 18 Adams J, Palombella VJ, Sausville EA. et al. Proteasome inhibitors: a novel class of potent and effective antitumor agents. Cancer Res 1999; 59: 2615-22.
    PubMedGoogle Scholar
  • 19 Lightcap ES, McCormack TA, Pien CS. et al. Proteasome inhibition measurements: clinical application. Clin Chem 2000; 46: 673-83.
    PubMedGoogle Scholar
  • 20 Chen J, Li Y, Wang L. et al. Therapeutic benefit of intracerebral transplantation of bone marrow stromal cells after cerebral ischemia in rats. J Neurol Sci 2001; 189: 49-57.
    CrossrefPubMedGoogle Scholar
  • 21 Schallert T, Whishaw IQ. Bilateral cutaneous stimulation of the somatosensory system in hemidecorticate rats. Behav Neurosci 1984; 98: 518-40.
    CrossrefPubMedGoogle Scholar
  • 22 Hernandez TD, Schallert T. Seizures and recovery from experimental brain damage. Exp Neurol 1988; 102: 318-24.
    CrossrefPubMedGoogle Scholar
  • 23 Swanson RA, Morton MT, Tsao-Wu G. et al. A semiautomated method for measuring brain infarct volume. J Cereb Blood Flow Metab 1990; 10: 290-3.
    CrossrefPubMedGoogle Scholar
  • 24 Zhang L, Zhang ZG, Zhang R. et al. Adjuvant treatment with a glycoprotein IIb/IIIa receptor inhibitor increases the therapeutic window for low-dose tissue plasminogen activator administration ina rat model of embolic stroke. Circulation 2003; 107: 2837-43.
    CrossrefPubMedGoogle Scholar
  • 25 Mun-Bryce S, Rosenberg GA. Matrix metalloproteinases in cerebrovascular disease. J Cereb Blood Flow Metab 1998; 18: 1163-72.
    CrossrefPubMedGoogle Scholar
  • 26 Uchinami H, Yamamoto Y, Kume M. et al. Effect of heat shock preconditioning on NF-kappaB/I-kappaB pathway during I/R injury of the rat liver. Am J Physiol Gastrointest Liver Physiol 2002; 282: G962-71.
    CrossrefPubMedGoogle Scholar
  • 27 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25: 402-8.
    CrossrefPubMedGoogle Scholar
  • 28 Zhang ZG, Zhang L, Tsang W. et al. Dynamic platelet accumulation at the site of the occluded middle cerebral artery and in downstream microvessels is associated with loss of microvascular integrity after embolic middle cerebral artery occlusion. Brain Res 2001; 912: 181-94.
    CrossrefPubMedGoogle Scholar
  • 29 Stringer HA, van Swieten P, Heijnen HF. et al. Plasminogen activator inhibitor-1 released from activated platelets plays a key role in thrombolysis resistance. Studies with thrombi generated in the Chandler loop. Arterioscler Thromb 1994; 14: 1452-8.
    CrossrefPubMedGoogle Scholar
  • 30 Ostrowska JK, Wojtukiewicz MZ, Chabielska E. et al. Proteasome inhibitor prevents experimental arterial thrombosis in renovascular hypertensive rats. Thromb Haemost 2004; 92: 171-7.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 31 Kolev K, Skopal J, Simon L. et al. Matrix metalloproteinase-9 expression in post-hypoxic human brain capillary endothelial cells: H2O2 as a trigger and NFkappaB as a signal transducer. Thromb Haemost 2003; 90: 528-37.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 32 Elliott PJ, Pien CS, McCormack TA. et al. Proteasome inhibition: A novel mechanism to combat asthma. J Allergy Clin Immunol 1999; 104: 294-300.
    CrossrefPubMedGoogle Scholar
  • 33 Justicia C, Panes J, Sole S. et al. Neutrophil infiltration increases matrix metalloproteinase-9 in the ischemic brain after occlusion/reperfusion of the middle cerebral artery in rats. J Cereb Blood Flow Metab 2003; 23: 1430-40.
    CrossrefPubMedGoogle Scholar
  • 34 Lo EH, Wang X, Cuzner ML. Extracellular proteolysis in brain injury and inflammation: role for plasminogen activators and matrix metalloproteinases. J Neurosci Res 2002; 69: 1-9.
    CrossrefPubMedGoogle Scholar
  • 35 Iadecola C, Pelligrino DA, Moskowitz MA. et al. Nitric oxide synthase inhibition and cerebrovascular regulation. J Cereb Blood Flow Metab 1994; 14: 175-92.
    CrossrefPubMedGoogle Scholar
  • 36 Podar K, Shringarpure R, Tai YT. et al. Caveolin-1 is required for vascular endothelial growth factor-triggered multiple myeloma cell migration and is targeted by bortezomib. Cancer Res 2004; 64: 7500-6.
    CrossrefPubMedGoogle Scholar
  • 37 Monaco C, Andreakos E, Kiriakidis S. et al. Canonical pathway of nuclear factor kappa B activation selectively regulates proinflammatory and prothrombotic responses in human atherosclerosis. Proc Natl Acad Sci US A 2004; 101: 5634-9.
    CrossrefPubMedGoogle Scholar
  • 38 Dechend R, Maass M, Gieffers J. et al. Chlamydia pneumoniae infection of vascular smooth muscle and endothelial cells activates NF-κB and induces tissue factor and PAI-1 expression: a potential link to accelerated arteriosclerosis. Circulation 1999; 100: 1369-73.
    CrossrefPubMedGoogle Scholar