This Article Full Text Full Text (PDF) All Versions of this Article: 35/5/1169    most recent 01.STR.0000125861.55804.f2v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Maier, C. M. Articles by Chan, P. H. Search for Related Content PubMed PubMed Citation Articles by Maier, C. M. Articles by Chan, P. H. Related Collections Animal models of human disease Ischemic biology - basic studies Acute Cerebral Hemorrhage Brain Circulation and Metabolism Other Stroke Treatment - Medical Other Vascular biology

(Stroke. 2004;35:1169.)
© 2004 American Heart Association, Inc.

Original Contributions

Matrix Metalloproteinase-9 and Myeloperoxidase Expression

Quantitative Analysis by Antigen Immunohistochemistry in a Model of Transient Focal Cerebral Ischemia

Carolina M. Maier, PhD; Lily Hsieh, BA; Fengshan Yu, MD; Paige Bracci, PhD Pak H. Chan, PhD

From the Department of Neurosurgery (C.M.M., L.H., F.Y., P.H.C.), Department of Neurology and Neurological Sciences and the Program in Neurosciences, Stanford University School of Medicine, Stanford, Calif; and the Department of Neurology (P.B.), University of California at San Francisco, Calif.

Correspondence to Dr Pak H. Chan, Neurosurgical Laboratories, Stanford University, 1201 Welch Rd, MSLS P304, Stanford, CA 94305-5487. E-mail phchan{at}stanford.edu

Background and Purpose— Expression of matrix metalloproteinases (MMPs), proteolytic enzymes that degrade extracellular proteins, is altered after ischemia/reperfusion injury and may contribute to blood–brain barrier (BBB) breakdown. Neutrophils, a source of reactive oxygen species and MMP-9, infiltrate damaged tissue 6 to 24 hours after ischemia and have also been implicated in delayed secondary tissue damage. Here we examined the spatial–temporal relation between MMP-9 expression and neutrophil infiltration after stroke.

Methods— Knockout mice containing 50% manganese superoxide dismutase activity (SOD2-KOs), which are more susceptible to ischemic damage than wild-type (WT) littermates, underwent quantitative antigen (MMP-9, myeloperoxidase) immunohistochemistry (24 and 72 hours) analysis and protein expression by Western blotting (6, 12, 24, 48, and 72 hours) after transient focal cerebral ischemia. BBB breakdown was determined by Evans blue extravasation.

Results— There was a clear spatial relation between MMP-9 expression and Evans blue extravasation. MMP-9–positive cell and vessel counts for SOD2-KOs (72 hours) were significantly different from SOD2-KO (24 hours, P=0.004), WT (24 hours, P=0.01), and WT (72 hours, P=0.007) mice. In contrast, MMP-9–positive neutrophil counts were comparatively low and did not differ by time or animal type. MMP-9 expression was biphasic in SOD2-KOs but not in WT littermates, with a significant increase observed 6 to 12 hours after ischemic insult and again at 48 to 72 hours. SOD2-KOs showed increased MMP-9 expression compared with WT littermates at all time points studied (P0.05).

Conclusions— In this model, neutrophils are not the primary source of MMP-9 protein and thus are unlikely the key contributor to BBB breakdown observed in SOD2-KOs.

Key Words: metalloproteinases • inflammation • cerebral ischemia, focal • blood-brain barrier • oxygen radical

This article has been cited by other articles:

				A. Rosell, E. Cuadrado, A. Ortega-Aznar, M. Hernandez-Guillamon, E. H. Lo, and J. Montaner 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 Stroke, April 1, 2008; 39(4): 1121 - 1126. [Abstract] [Full Text] [PDF]

				E. M. Hurt, S. B. Thomas, B. Peng, and W. L. Farrar Integrated molecular profiling of SOD2 expression in multiple myeloma Blood, May 1, 2007; 109(9): 3953 - 3962. [Abstract] [Full Text] [PDF]

				B. W. McColl, N. J. Rothwell, and S. M. Allan Systemic Inflammatory Stimulus Potentiates the Acute Phase and CXC Chemokine Responses to Experimental Stroke and Exacerbates Brain Damage via Interleukin-1- and Neutrophil-Dependent Mechanisms J. Neurosci., April 18, 2007; 27(16): 4403 - 4412. [Abstract] [Full Text] [PDF]

				P. Yan, X. Hu, H. Song, K. Yin, R. J. Bateman, J. R. Cirrito, Q. Xiao, F. F. Hsu, J. W. Turk, J. Xu, et al. Matrix Metalloproteinase-9 Degrades Amyloid-beta Fibrils in Vitro and Compact Plaques in Situ J. Biol. Chem., August 25, 2006; 281(34): 24566 - 24574. [Abstract] [Full Text] [PDF]

				M. Ning, K. L. Furie, W. J. Koroshetz, H. Lee, M. Barron, M. Lederer, X. Wang, M. Zhu, A. G. Sorensen, E. H. Lo, et al. Association between tPA therapy and raised early matrix metalloproteinase-9 in acute stroke Neurology, May 23, 2006; 66(10): 1550 - 1555. [Abstract] [Full Text] [PDF]

				A. S. Haqqani, M. Nesic, E. Preston, E. Baumann, J. Kelly, and D. Stanimirovic Characterization of vascular protein expression patterns in cerebral ischemia/reperfusion using laser capture microdissection and ICAT-nanoLC-MS/MS FASEB J, November 1, 2005; 19(13): 1809 - 1821. [Abstract] [Full Text] [PDF]

				J. M. Gidday, Y. G. Gasche, J.-C. Copin, A. R. Shah, R. S. Perez, S. D. Shapiro, P. H. Chan, and T. S. Park Leukocyte-derived matrix metalloproteinase-9 mediates blood-brain barrier breakdown and is proinflammatory after transient focal cerebral ischemia Am J Physiol Heart Circ Physiol, August 1, 2005; 289(2): H558 - H568. [Abstract] [Full Text] [PDF]

				J. Wang and S. E. Tsirka Neuroprotection by inhibition of matrix metalloproteinases in a mouse model of intracerebral haemorrhage Brain, July 1, 2005; 128(7): 1622 - 1633. [Abstract] [Full Text] [PDF]