Plasma and Brain Matrix Metalloproteinase-9 After Acute Focal Cerebral Ischemia in Rats

Background and Purpose— Plasma levels of matrix metalloproteinase-9 (MMP-9) have been proposed to be a useful biomarker for assessing pathological events in brain. Here, we examined the temporal profiles of MMP-9 in blood and brain using a rat model of acute focal cerebral ischemia. Methods— Plasma and brain levels of MMP-2 and MMP-9 were quantified at 3, 6, 12, and 24 hours after permanent middle cerebral artery occlusion in male Sprague-Dawley rats. Infarct volumes at 24 hours were confirmed with 2,3,5-triphenyl-tetrazolium-chloride staining. Results— In plasma, zymographic bands were detected between 70 and 95 kDa corresponding to pro-MMP-2, pro-MMP-9, and activated MMP-9. A higher 135-kDa band was also seen that is likely to be NGAL-conjugated MMP-9. After ischemia, there were no significant changes in pro-MMP-2, but plasma levels of pro-MMP-9 steadily increased over the course of 24 hours. Activated MMP-9 levels in plasma were significantly elevated only at 24 hours. Plasma NGAL-MMP-9 complexes showed a transient elevation between 3 to 6 hours, after which levels decreased back down to pre-ischemic baselines. In brain homogenates, pro-MMP-2, pro-MMP-9, and activated MMP-9 were seen but no NGAL-MMP-9 bands were detected. Compared to the contralateral hemisphere, MMP-2 and MMP-9 levels in ischemic brain progressively increased over the course of 24 hours. Overall levels of MMP-9 in plasma and brain were significantly correlated, especially at 24 hours. Plasma levels of pro-MMP-9 at 24 hours were correlated with final infarct volumes. Conclusions— Elevated plasma levels of MMP-9 appear to be correlated with brain levels within 24 hours of acute cerebral ischemia in rats. Further investigation into clinical profiles of MMP-9 in acute stroke patients may be useful.

[1]  Peter Sandercock,et al.  Blood Biomarkers in the Diagnosis of Ischemic Stroke: A Systematic Review , 2008, Stroke.

[2]  E. Lo,et al.  MMP-9–Positive Neutrophil Infiltration Is Associated to Blood–Brain Barrier Breakdown and Basal Lamina Type IV Collagen Degradation During Hemorrhagic Transformation After Human Ischemic Stroke , 2008, Stroke.

[3]  M. Corasaniti,et al.  Brain regional and cellular localization of gelatinase activity in rat that have undergone transient middle cerebral artery occlusion , 2008, Neuroscience.

[4]  S. Heiland,et al.  Minocycline and hypothermia for reperfusion injury after focal cerebral ischemia in the rat—Effects on BBB breakdown and MMP expression in the acute and subacute phase , 2008, Brain Research.

[5]  Jialiang Hu,et al.  Matrix metalloproteinase inhibitors as therapy for inflammatory and vascular diseases , 2007, Nature Reviews Drug Discovery.

[6]  Jeffrey F. Thompson,et al.  Matrix Metalloproteinase-Mediated Disruption of Tight Junction Proteins in Cerebral Vessels is Reversed by Synthetic Matrix Metalloproteinase Inhibitor in Focal Ischemia in Rat , 2007, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[7]  Qing Wang,et al.  The inflammatory response in stroke , 2007, Journal of Neuroimmunology.

[8]  E. Lo,et al.  Neurovascular Proteases in Brain Injury, Hemorrhage and Remodeling After Stroke , 2007, Stroke.

[9]  K. Furie,et al.  Association between tPA therapy and raised early matrix metalloproteinase-9 in acute stroke , 2006, Neurology.

[10]  V. Yong,et al.  Metalloproteinases: Mediators of Pathology and Regeneration in the CNS , 2005, Nature Reviews Neuroscience.

[11]  S. Koh,et al.  Effect of 3-aminobenzamide, PARP inhibitor, on matrix metalloproteinase-9 level in plasma and brain of ischemic stroke model. , 2005, Toxicology.

[12]  R. Kauppinen,et al.  Minocycline Protects against Permanent Cerebral Ischemia in Wild Type but Not in Matrix Metalloprotease-9-Deficient Mice , 2005, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[13]  E. Lo,et al.  Induction of Caspase-Mediated Cell Death by Matrix Metalloproteinases in Cerebral Endothelial Cells after Hypoxia—Reoxygenation , 2004, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[14]  Alex Rovira,et al.  Plasmatic Level of Neuroinflammatory Markers Predict the Extent of Diffusion-Weighted Image Lesions in Hyperacute Stroke , 2003, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[15]  J. Koziol,et al.  Profiles of matrix metalloproteinases, their inhibitors, and laminin in stroke patients: influence of different therapies. , 2003, Stroke.

[16]  K. Arai,et al.  Essential role for ERK mitogen‐activated protein kinase in matrix metalloproteinase‐9 regulation in rat cortical astrocytes , 2003, Glia.

[17]  J. Serena,et al.  Plasma Metalloproteinase-9 Concentration Predicts Hemorrhagic Transformation in Acute Ischemic Stroke , 2003, Stroke.

[18]  E. Lo,et al.  Blood-Brain Barrier Disruption and Matrix Metalloproteinase-9 Expression During Reperfusion Injury: Mechanical Versus Embolic Focal Ischemia in Spontaneously Hypertensive Rats , 2002, Stroke.

[19]  Jiankun Cui,et al.  S-Nitrosylation of Matrix Metalloproteinases: Signaling Pathway to Neuronal Cell Death , 2002, Science.

[20]  J. Arenillas,et al.  Matrix Metalloproteinase Expression After Human Cardioembolic Stroke: Temporal Profile and Relation to Neurological Impairment , 2001, Stroke.

[21]  Inge Nelissen,et al.  Gelatinase B functions as regulator and effector in leukocyte biology , 2001, Journal of leukocyte biology.

[22]  M. Fini,et al.  Role for Matrix Metalloproteinase 9 after Focal Cerebral Ischemia: Effects of Gene Knockout and Enzyme Inhibition with BB-94 , 2000, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[23]  J. Koziol,et al.  Matrix Metalloproteinases Increase Very Early during Experimental Focal Cerebral Ischemia , 1999, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[24]  G. Rosenberg,et al.  Matrix Metalloproteinases in Cerebrovascular Disease , 1998, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[25]  G. Rosenberg,et al.  Matrix metalloproteinases and TIMPs are associated with blood-brain barrier opening after reperfusion in rat brain. , 1998, Stroke.

[26]  F. Barone,et al.  Matrix metalloproteinase expression increases after cerebral focal ischemia in rats: inhibition of matrix metalloproteinase-9 reduces infarct size. , 1998, Stroke.

[27]  H. Sengeløv,et al.  Isolation and primary structure of NGAL, a novel protein associated with human neutrophil gelatinase. , 1993, The Journal of biological chemistry.

[28]  Teng-Nan Lin,et al.  Effect of Brain Edema on Infarct Volume in a Focal Cerebral Ischemia Model in Rats , 1993, Stroke.