Molecular mechanisms for NG-nitro-L-arginine methyl ester action against cerebral ischemia–reperfusion injury-induced blood–brain barrier dysfunction

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Abstract

Background: Ischemic stroke, an acute neurological injury lacking an effective therapy, is a leading cause of death worldwide. The unmet need in stroke research is to identify viable therapeutic targets and to understand their interplay during cerebral ischemia-reperfusion (I/R) injury.

Objective: To explore the protective effects and molecular mechanism of NG-nitro-L-arginine methyl ester (L-NAME) in cerebral ischemia-reperfusion injury-induced blood-brain barrier (BBB) dysfunction.

Methods: Two hundred fifty-six rats were randomly assigned to a sham operation group, I/R group, and I/R with L-NAME treatment group. Brain water content was determined by calculating dry/wet weight. The permeability of the BBB was observed using an electron microscope and by determining the Evans Blue leakage from brain tissue on the ischemic side. The expression of brain MMP-9 and GFAP was determined using an immunohistochemical method. The expression of ZO-1 protein was determined by western blotting.

Results: We found that L-NAME remarkably attenuated the permeability of the BBB after I/R as assessed by Evans Blue leakage and brain water content (p < 0.05). This was further confirmed by examination of the ultrastructural morphology of the BBB using a transmission electron microscope. Furthermore, we found that expression of the zonae occludens-1 (ZO-1) was decreased in endothelial cells, and expression of MMP-9 and GFAP was increased in the basement membrane and astrocyte end-feet in vehicle control groups, respectively, but these changes could be prevented by L-NAME pretreatment.

Conclusion: These results suggested that the neuroprotective effects of L-NAME against BBB damage induced by I/R might be related to the upregulation of tight junction proteins and inhibition of MMP-9 and GFAP expression. L-NAME can be used as a potential MMP-9-based multiple targeting therapeutic strategy in cerebral I/R injury.

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  • 1. Li M Qu YZ Zhao ZW Wu SX Liu YY Wei XY et al. Astragaloside IV protects against focal cerebral ischemia/reperfusion injury correlating to suppression of neutrophils adhesion-related molecules. Neurochem Int. 2012; 60:458-65.

  • 2. Roger VL Go AS Lloyd-Jones DM Benjamin EJ Berry JD Borden WB et al. Heart disease and stroke statistics-2012 update: a report from the American Heart Association. Circulation. 2012; 125: e2-e220.

  • 3. Datta A Jingru Q Khor TH Teo MT Heese K Sze SK. Quantitative neuroproteomics of an in vivo rodent model of focal cerebral ischemia/reperfusion injury reveals a temporal regulation of novel pathophysiological molecular markers. J Proteome Res. 2011; 10: 5199-213.

  • 4. Wang Y Liao X Zhao X Wang DZ Wang C Nguyen-Huynh MN et al. Using recombinant tissue plasminogen activator to treat acute ischemic stroke in China: analysis of the results from the Chinese National Stroke Registry (CNSR). Stroke. 2011; 42: 1658-64.

  • 5. Tsubota H Marui A Esaki J Bir SC Ikeda T Sakata R. Remote postconditioning may attenuate ischaemia- reperfusion injury in the murine hindlimb through adenosine receptor activation. Eur J Vasc Endovasc Surg. 2010; 40:804-9.

  • 6. Kago T Takagi N Date I Takenaga Y Takagi K Takeo S. Cerebral ischemia enhances tyrosine phosphorylation of occludin in brain capillaries. Biochem Biophys Res Commun. 2006; 339:1197-203.

  • 7. Jiao H Wang Z Liu Y Wang P Xue Y. Specific role of tight junction proteins claudin-5 occludin and ZO-1 of the blood-brain barrier in a focal cerebral ischemic insult. J Mol Neurosci. 2011; 44:130-9.

  • 8. Rosenberg GA Estrada EY Dencoff JE. Matrix metalloproteinases and TIMPs are associated with blood-brain barrier opening after reperfusion in rat brain. Stroke. 1998; 29:2189-95.

  • 9. Fukuda S Fini CA Mabuchi T Koziol JA Eggleston LL Jr del Zoppo GJ. Focal cerebral ischemia induces active proteases that degrade microvascular matrix. Stroke. 2004; 35:998-1004.

  • 10. Bauer AT B ⃞rgers HF Rabie T Marti HH. Matrix metalloproteinase-9 mediates hypoxia-induced vascular leakage in the brain via tight junction rearrangement. J Cereb Blood Flow Metab. 2010; 30: 837-48.

  • 11. Dejonckheere E Vandenbroucke RE Libert C. Matrix metalloproteinases as drug targets in ischemia/ reperfusion injury. Drug Discov Today. 2011; 16: 762-78.

  • 12. Pascual O Casper KB Kubera C Zhang J Revilla- Sanchez R Sul JY et al. Astrocytic purinergic signaling coordinates synaptic networks. Science. 2005; 310: 113-6.

  • 13. Wang W Redecker C Yu ZY Xie MJ Tian DS Zhang L et al. Rat focal cerebral ischemia induced astrocyte proliferation and delayed neuronal death are attenuated by cyclin-dependent kinase inhibition. J Clin Neurosci. 2008; 15:278-85.

  • 14. Gursoy-Ozdemir Y Bolay H Sariba_ O Dalkara T. Role of endothelial nitric oxide generation and peroxynitrite formation in reperfusion injury after focal cerebral ischemia. Stroke. 2000; 31:1974-80; discussion 1981.

  • 15. Gursoy-Ozdemir Y Can A Dalkara T. Reperfusioninduced oxidative/nitrative injury to neurovascular unit after focal cerebral ischemia. Stroke. 2004; 35: 1449-53.

  • 16. Gu Y Zheng G Xu M Li Y Chen X Zhu W et al. Caveolin-1 regulates nitric oxide-mediated matrix metalloproteinases activity and blood-brain barrier permeability in focal cerebral ischemia and reperfusion injury. J Neurochem. 2012; 120:147-56.

  • 17. Stevanovic ID Jovanovic MD Colic M Jelenkovic A Bokonjic D Ninkovic M et al. N-nitro-L-arginine methyl ester influence on aluminium toxicity in the brain. Folia Neuropathol. 2011; 49:219-29.

  • 18. Longa EZ Weinstein PR Carlson S Cummins R. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke. 1989; 20:84-91.

  • 19. Vakili A Kataoka H Plesnila N. Role of arginine vasopressin V1 and V2 receptors for brain damage after transient focal cerebral ischemia. J Cereb Blood Flow Metab. 2005; 25:1012-9.

  • 20. Belayev L Busto R Zhao W Ginsberg MD. Quantitative evaluation of blood-brain barrier permeability following middle cerebral artery occlusion in rats. Brain Res. 1996; 739:88-96.

  • 21. Yu F Kamada H Niizuma K Endo H Chan PH. Induction of MMP-9 expression and endothelial injury by oxidative stress after spinal cord injury. J Neurotrauma. 2008; 25:184-95.

  • 22. Date I Takagi N Takagi K Tanonaka K Funakoshi H Matsumoto K et al. Hepatocyte growth factor attenuates cerebral ischemia- induced increase in permeability of the blood-brain barrier and decreases in expression of tight junctional proteins in cerebral vessels. Neurosci Lett. 2006; 407:141-5.

  • 23. Qu YZ Li M Zhao YL Zhao ZW Wei XY Liu JP et al. Astragaloside IV attenuates cerebral ischemia- reperfusion-induced increase in permeability of the blood-brain barrier in rats. Eur J Pharmacol. 2009; 606: 137-41.

  • 24. Jung JE Kim GS Chen H Maier CM Narasimhan P Song YS et al. Reperfusion and neurovascular dysfunction in stroke: from basic mechanisms to potential strategies for neuroprotection. Mol Neurobiol. 2010; 41:172-9.

  • 25. Glaser N. Cerebral injury and cerebral edema in children with diabetic ketoacidosis: could cerebral ischemia and reperfusion injury be involved? Pediatr Diabetes. 2009; 10:534-41.

  • 26. Ballabh P Braun A Nedergaard M. The blood-brain barrier: an overview: structure regulation and clinical implications. Neurobiol Dis. 2004; 16:1-13.

  • 27. Dohgu S Takata F Yamauchi A Nakagawa S Egawa T Naito M et al. Brain pericytes contribute to the induction and up-regulation of blood-brain barrier functions through transforming growth factor- â production. Brain Res. 2005; 1038:208-15.

  • 28. Pun PB Lu J Moochhala S. Involvement of ROS in BBB dysfunction. Free Radic Res. 2009; 43:348-64.

  • 29. Zan L Wu H Jiang J Zhao S Song Y Teng G et al. Temporal profile of Src SSeCKS and angiogenic factors after focal cerebral ischemia: correlations with angiogenesis and cerebral edema. Neurochem Int. 2011; 58:872-9.

  • 30. Ortinski PI Dong J Mungenast A Yue C Takano H Watson DJ et al. Selective induction of astrocytic gliosis generates deficits in neuronal inhibition. Nat Neurosci. 2010; 13:584-91.

  • 31. Cheung WM Wang CK Kuo JS Lin TN. Changes in the level of glial fibrillary acidic protein (GFAP) after mild and severe focal cerebral ischemia. Chin J Physiol. 1999; 42:227-35.

  • 32. Pfefferkorn T Rosenberg GA. Closure of the bloodbrain barrier by matrix metalloproteinase inhibition reduces rtPA-mediated mortality in cerebral ischemia with delayed reperfusion. Stroke. 2003; 34: 2025-30.

  • 33. Horstmann S Kalb P Koziol J Gardner H Wagner S. Profiles of matrix metalloproteinases their inhibitors and laminin in stroke patients: influence of different therapies. Stroke. 2003; 34:2165-70.

  • 34. Suofu Y Clark JF Broderick JP Kurosawa Y Wagner KR Lu A. Matrix metalloproteinase-2 or -9 deletions protect against hemorrhagic transformation during early stage of cerebral ischemia and reperfusion. Neuroscience. 2012; 212:180-9.

  • 35. Ryang Y-M Dang J Kipp M Petersen K-U Fahlenkamp AV Gempt J et al. Solulin reduces infarct volume and regulates gene-expression in transient middle cerebral artery occlusion in rats. BMC Neurosci. 2011; 12:113.

  • 36. Wu Y Wang YP Guo P Ye XH Wang J Yuan SY et al. A lipoxin A4 analog ameliorates blood-brain barrier dysfunction and reduces MMP-9 expression in a rat model of focal cerebral ischemia-reperfusion injury. J Mol Neurosci. 2012; 46: 483-91.

  • 37. Tai S-H Chen H-Y Lee E-J Chen T-Y Lin H-W Hung Y-C et al. Melatonin inhibits postischemic matrix metalloproteinase-9 (MMP-9) activation via dual modulation of plasminogen/plasmin system and endogenous MMP inhibitor in mice subjected to transient focal cerebral ischemia. J Pineal Res. 2010; 49:332-41.

  • 38. Wang Z Xue Y Jiao H Liu Y Wang P. Doxycyclinemediated protective effect against focal cerebral ischemia-reperfusion injury through the modulation of tight junctions and PKCä signaling in rats. J Mol Neurosci. 2012; 47:89-100.

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