PDF herunterladen
Thromb Haemost 2010; 103(05): 891-900
DOI: 10.1160/TH09-07-0500
Theme Issue Article
Schattauer GmbH

Different therapy options protecting microvasculature after experimental cerebral ischaemia and reperfusion

Dorothe Burggraf
1   Department of Neurology, Ludwig-Maximilians University, Klinikum Großhadern, Munich, Germany
,
Milan R. Vosko
2   Department of Neurology and Psychiatry, Linz, Austria
,
Marion Schubert
1   Department of Neurology, Ludwig-Maximilians University, Klinikum Großhadern, Munich, Germany
,
Jean-Marie Stassen
3   Thrombogenics NV, Leuven, Belgium
,
Gerhard F. Hamann
4   Department of Neurology, HSK Dr. Horst-Schmidt-Klinik, Wiesbaden, Germany
› Institutsangaben Financial support: This study was supported by Thrombogenics NV, a Belgian Biopharmaceutical Development Company. ThrombogenicsNV is the license holder of the use of microplasmin for the treatment of ischaemic stroke. The study was supported by a Marie-Curie individual fellowship of the European Community (QLRI-CT-2001–51077; M.R.V.).
Weitere Informationen

Publikationsverlauf

Received: 31. Juli 2009

Accepted after major revision: 11. März 2009

Publikationsdatum:
22. November 2017 (online)

Auch verfügbar auf
    Lizenzen und Reprints

Summary

Recombinant tissue plasminogen activator (rt-PA) is successfully used in human stroke, but often shows serious drawbacks. To find an alternative, we hypothesised that the novel thrombolytic microplasmin would have fewer adverse effects on haemoglobin extravasation and microvascular damage compared with the effects of rt-PA and tenecteplase (TNK). A constant period of ischaemia (3 hours) was induced in a rat suture model followed by reperfusion (24 hours). Mikroplasmin (10 mg/kg), TNK (5 mg/kg), rt-PA (9 mg/kg) and saline (control), were administered. The volume of the ischaemic lesion was calculated, the loss of collagen type IV and the extravasation of haemoglobin were quantified by Western blotting. The matrix-metalloproteinases 2 and 9 (MMP-2/-9) were quantified by zymography and their endogenous tissue inhibitors (TIMPs) were analysed by reverse zymography. Microplasmin treatment caused the lowest volume of the ischaemic lesion (51.0 ± 22.6 mm3) compared with control (167.3 ± 13.1 mm3; p<0.05). The content of col-lagen type IV was significantly increased and haemoglobin extravasation reduced (154 ± 24%; p<0.05) compared with control (442 ± 124%); MMP-2/-9 and the corresponding TIMPs remained unchanged. In comparison, TNK did not significantly reduce basal lamina damage and caused the highest extravasation. MMP-2/-9 were severely increased after TNK treatment (p<0.05). Thus, the balance between MMPs and TIMPs was shifted toward the inhibitory side with TNK. Microplasmin had a protective effect on the microvascular basal lamina and blood-brain barrier, whereas TNK was significantly disadvantageous from the viewpoint of ischaemic damage. Microplasmin also appears to be safer than other PAs in terms of damage to the microvasculature associated with thrombolytic therapy of ischaemic stroke.

Keywords

Cerebral ischaemia - microplasmin - MMPs - plasminogen activator - rat - rt-PA - tenecteplase
 

  • References

  • 1 The national institute of neurological disorders and stroke rtPA Stroke Study Group.. Tissue plasminogen activator for acute ischemic Stroke. N Engl J Med 1995; 333: 1581-1588.
    CrossrefPubMedGoogle Scholar
  • 2 Benchenane K, Berezowski V, Fernandez-Monreal M. et al. Oxygen glucose deprivation switches the transport of tPA across the blood-brain barrier from an LRP-dependent to an increased LRP-independent process. Stroke 2005; 36: 1065-1070.
    CrossrefPubMedGoogle Scholar
  • 3 Jespers L, Vanwetswinkel S, Lijnen HR. et al. Structural and functional basis of plasminogen activation by staphylokinase. Thromb Haemost 1999; 81: 479-485.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 4 Gibson CM, Cannon CP, Murphy SA. et al. Weight-adjusted dosing of TNK-tissue plasminogen activator and its relation to angiographic outcomes in the thrombolysis in myocardial infarction 10B trial. TIMI 10B Investigators. Am J Cardiol 1999; 84: 976-980.
    CrossrefPubMedGoogle Scholar
  • 5 Davydov L, Cheng JW. Tenecteplase: a review. Clin Ther 2001; 23: 982-997.
    CrossrefPubMedGoogle Scholar
  • 6 Shi GY, Wu HL. Isolation and characterization of microplasminogen. A low molecular weight form of plasminogen. J Biol Chem 1988; 263: 17071-17075.
    PubMedGoogle Scholar
  • 7 Lapchak PA. Development of thrombolytic therapy for stroke: a perspective. Expert Opin Investig Drugs 2002; 01: 1623-1632.
    PubMedGoogle Scholar
  • 8 Nagai N, Demarsin E, Van Hoef B. et al. Recombinant human microplasmin: production and potential therapeutic properties. J Thromb Haemost 2003; 01: 307-313.
    CrossrefPubMedGoogle Scholar
  • 9 Hamann GF, Okada Y, del Zoppo GJ. Hemorrhagic transformation and microvascular integrity during focal cerebral ischemia/reperfusion. J Cereb Blood Flow Metab 1996; 16: 1373-1378.
    CrossrefPubMedGoogle Scholar
  • 10 Vosko MR, Burggraf D, Liebetrau M. et al. Influence of the duration of ischemia and reperfusion on infarct volume and microvascular damage in mice. Neurol Res 2006; 28: 200-205.
    CrossrefPubMedGoogle Scholar
  • 11 Hamann GF, Okada Y, Fitridge R. et al. Microvascular basal lamina antigens disappear during cerebral ischemia and reperfusion. Stroke 1995; 26: 2120-2126.
    CrossrefPubMedGoogle Scholar
  • 12 Hamann GF, Liebetrau M, Martens HK. Microvascular basal lamina injury after experimental focal cerebral ischemia and reperfusion in the rat. J Cereb Blood Flow Metab 2002; 22: 526-533.
    CrossrefPubMedGoogle Scholar
  • 13 Mignatti P. Extracellular matrix remodeling by metalloproteinases and plasminogen activators. Kidney Int Suppl 1995; 49: S12-14.
    PubMedGoogle Scholar
  • 14 Burggraf D, Martens HK, Jager G. et al. Recombinant human tissue plasminogen activator protects the basal lamina in experimental focal cerebral ischemia. Thromb Haemost 2003; 89: 1072-1080.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 15 Van de Werf F, Cannon CP, Luyten A. et al. Safety assessment of single-bolus administration of TNK tissue-plasminogen activator in acute myocardial infarction: the ASSENT-1 trial. Am Heart J 1999; 137: 786-791.
    CrossrefPubMedGoogle Scholar
  • 16 Paxinos G, Watson C. The rat brain in stereotaxic coordinates. 1998. San Diego; Academic Press.:
    Google Scholar
  • 17 Burggraf D, Martens HK, Liebetrau M. et al. A new approach to reduce the number of animals used in experimental focal cerebral ischemia models. Neurosci Lett 2005; 386: 88-93.
    CrossrefPubMedGoogle Scholar
  • 18 Miyazawa T, Bonnekoh P, Hossmann KA. Temperature effect on immunostaining of microtubule-associated protein 2 and synaptophysin after 30 minutes of fore-brain ischemia in rat. Acta Neuropathol 1993; 85: 526-532.
    PubMedGoogle Scholar
  • 19 Kloss CU, Thomassen N, Fesl G. et al. Tissue-saving infarct volumetry using histochemistry validated by MRI in rat focal ischemia. Neurol Res 2002; 24: 713-718.
    CrossrefPubMedGoogle Scholar
  • 20 Ogata Y, Itoh Y, Nagase H. Steps involved in activation of the pro-matrix metalloproteinase 9 (progelatinase B)-tissue inhibitor of metalloproteinases-1 complex by 4-aminophenylmercuric acetate and proteinases. J Biol Chem 1995; 270: 18506-18511.
    CrossrefPubMedGoogle Scholar
  • 21 Hacke W, Donnan G, Fieschi C. et al. ATLANTIS Trials Investigators; ECASS Trials Investigators; NINDS rt-PA Study Group Investigators. Association of outcome with early stroke treatment: pooled analysis of ATLANTIS, ECASS, and NINDS rt-PA stroke trials. Lancet 2004; 363: 768-774.
    CrossrefPubMedGoogle Scholar
  • 22 Kilic E, Bahr M, Hermann DM. Effects of recombinant tissue plasminogen activator after intraluminal thread occlusion in mice: role of hemodynamic alterations. Stroke 2001; 32: 2641-2647.
    CrossrefPubMedGoogle Scholar
  • 23 Burggraf D, Martens HK, Wunderlich N. et al. Rt-PA causes a significant increase in endogenous u-PA during experimental focal cerebral ischemia. Eur J Neurosci 2004; 20: 2903-2908.
    CrossrefPubMedGoogle Scholar
  • 24 Kilic E, Hermann DM, Hossmann KA. Recombinant tissue plasminogen activator reduces infarct size after reversible thread occlusion of middle cerebral artery in mice. Neuroreport 1999; 10: 107-111.
    CrossrefPubMedGoogle Scholar
  • 25 Tabrizi P, Wang L, Seeds N. et al. Tissue plasminogen activator (tPA) deficiency exacerbates cerebrovascular fibrin deposition and brain injury in a murine stroke model: studies in tPA-deficient mice and wild-type mice on a matched genetic background. Arterioscler Thromb Vasc Biol 1999; 19: 2801-2806.
    CrossrefPubMedGoogle Scholar
  • 26 Wang YF, Tsirka SE, Strickland S. et al. Tissue plasminogen activator (tPA) increases neuronal damage after focal cerebral ischemia in wild-type and tPA-deficient mice. Nat Med 1998; 04: 228-231.
    CrossrefPubMedGoogle Scholar
  • 27 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.
    CrossrefPubMedGoogle Scholar
  • 28 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: 10898-10907.
    CrossrefPubMedGoogle Scholar
  • 29 Nagai N, Zhao BQ, Suzuki Y. et al. Tissue-type plasminogen activator has paradoxical roles in focal cerebral ischemic injury by thrombotic middle cerebral artery occlusion with mild or severe photochemical damage in mice. J Cereb Blood Flow Metab 2002; 22: 648-651.
    CrossrefPubMedGoogle Scholar
  • 30 Lo EH, Broderick JP, Moskowitz MA. tPA and proteolysis in the neurovascular unit. Stroke 2004; 35: 354-356.
    CrossrefPubMedGoogle Scholar
  • 31 Trinkl A, Vosko MR, Wunderlich N. et al. Pravastatin reduces microvascular basal lamina damage following focal cerebral ischemia and reperfusion. Eur J Neurosci 2006; 24: 520-526.
    CrossrefPubMedGoogle Scholar
  • 32 Montaner J, Molina CA, Monasterio J. et al. Matrix metalloproteinase-9 pretreatment level predicts intracranial hemorrhagic complications after thrombolysis in human stroke. Circulation 2003; 107: 598-603.
    CrossrefPubMedGoogle Scholar
  • 33 Refino CJ, Paoni NF, Keyt BA. et al. A variant of t-PA (T103N, KHRR 296–299 AAAA) that, by bolus, has increased potency and decreased systemic activation of plasminogen. Thromb Haemost 1993; 70: 313-319.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 34 Collen D, Stassen JM, Yasuda T. et al. Comparative thrombolytic properties of tissue-type plasminogen activator and of a plasminogen activator inhibitor-1-resistant glycosylation variant, in a combined arterial and venous thrombosis model in the dog. Thromb Haemost 1994; 72: 98-104.
    PubMedGoogle Scholar
  • 35 Chapman DF, Lyden P, Lapchak PA. et al. Comparison of TNK with wild-type tissue plasminogen activator in a rabbit embolic stroke model. Stroke 2001; 32: 748-752.
    CrossrefPubMedGoogle Scholar
  • 36 Haley EC, Lyden PD, Johnston KC. et al. TNK in Stroke Investigators. A pilot dose-escalation safety study of tenecteplase in acute ischemic stroke. Stroke 2005; 36: 607-612.
    CrossrefPubMedGoogle Scholar
  • 37 Zhang L, Zhang ZG, Zhang C. et al. Intravenous administration of a GPIIb/IIIa receptor antagonist extends the therapeutic window of intra-arterial tenecteplase-tissue plasminogen activator in a rat stroke model. Stroke 2004; 35: 2890-2895.
    CrossrefPubMedGoogle Scholar
  • 38 Nagai N, De Mol M, Van Hoef B. et al. Depletion of circulating a2-antiplasmin by intravenous plasmin or immunoneutralization reduces focal cerebral ischemic injury in the absence of arterial recanalization. Blood 2001; 97: 3086-3092.
    CrossrefPubMedGoogle Scholar
  • 39 Suzuki Y, Chen F, Ni Y, Marchal G. et al. Microplasmin reduces ischemic brain damage and improves neurological function in a rat stroke model monitored with MRI. Stroke 2004; 35: 2402-2406.
    CrossrefPubMedGoogle Scholar
  • 40 Chen F, Suzuki Y, Nagai N. et al. Microplasmin and tissue plasminogen activator: comparison of therapeutic effects in rat stroke model at multiparametric MR imaging. Radiology 2007; 244: 429-438.
    CrossrefPubMedGoogle Scholar
  • 41 Kano T, Katayama Y, Tejima E. et al. Hemorrhagic transformation after fibrinolytic therapy with tissue plasminogen activator in a rat thromboembolic model of stroke. Brain Res 2000; 854: 245-248.
    CrossrefPubMedGoogle Scholar
  • 42 Verstraete M. Third-generation thrombolytic drugs. Am J Med 2000; 109: 52-58.
    CrossrefPubMedGoogle Scholar
  • 43 Khatri P, Neff J, Broderick JP. et al. IMS-I Investigators. Revascularization end points in stroke interventional trials: recanalization versus reperfusion in IMS-I. Stroke 2005; 36: 2400-2403.
    CrossrefPubMedGoogle Scholar