Post-transcriptional inactivation of matrix metalloproteinase-12 after focal cerebral ischemia attenuates brain damage
Bharath Chelluboina, Aditi Warhekar, Matt Dillard, Jeffrey D Klopfenstein, David M Pinson, David Z Wang, Krishna Kumar Veeravalli, Bharath Chelluboina, Aditi Warhekar, Matt Dillard, Jeffrey D Klopfenstein, David M Pinson, David Z Wang, Krishna Kumar Veeravalli
Abstract
This study highlights the possible pathological role of MMP-12 in the context of ischemic stroke. Male rats were subjected to a two-hour middle cerebral artery occlusion (MCAO) procedure. MMP-12 shRNA expressing plasmid formulation was administered to these rats twenty-four hours after reperfusion. The results showed a predominant upregulation of MMP-12 (approximately 47, 58, 143, and 265 folds on days 1, 3, 5, 7 post-ischemia, respectively) in MCAO subjected rats. MMP-12 expression was localized to neurons, oligodendrocytes and microglia, but not astrocytes. Transcriptional inactivation of MMP-12 significantly reduced the infarct size. The percent infarct size was reduced from 62.87±4.13 to 34.67±5.39 after MMP-12 knockdown compared to untreated MCAO subjected rats. Expression of myelin basic protein was increased, and activity of MMP-9 was reduced in ischemic rat brains after MMP-12 knockdown. Furthermore, a significant reduction in the extent of apoptosis was noticed after MMP-12 knockdown. TNFα expression in the ipsilateral regions of MCAO-subjected rats was reduced after MMP-12 knockdown in addition to the reduced protein expression of apoptotic molecules that are downstream to TNFα signaling. Specific knockdown of MMP-12 after focal cerebral ischemia offers neuroprotection that could be mediated via reduced MMP-9 activation and myelin degradation as well as inhibition of apoptosis.
Conflict of interest statement
The authors declare no competing financial interests.
Figures
References
- Banerjee S., Williamson D., Habib N., Gordon M. & Chataway J. Human stem cell therapy in ischaemic stroke: a review. Age Ageing 40, 7–13 (2011).
- del Zoppo G. J. tPA: a neuron buster, too? Nat. Med. 4, 148–150 (1998).
- Hacke W. et al. Thrombolysis in acute ischemic stroke: controlled trials and clinical experience. Neurology 53, S3–14 (1999).
- Wang X. et al. Lipoprotein receptor-mediated induction of matrix metalloproteinase by tissue plasminogen activator. Nat. Med. 9, 1313–1317 (2003).
- Gasche Y. et al. Early appearance of activated matrix metalloproteinase-9 after focal cerebral ischemia in mice: a possible role in blood-brain barrier dysfunction. J. Cereb. Blood Flow Metab. 19, 1020–1028 (1999).
- Heo J. H. et al. Matrix metalloproteinases increase very early during experimental focal cerebral ischemia. J. Cereb. Blood Flow Metab. 19, 624–633 (1999).
- Mun-Bryce S. & Rosenberg G. A. Matrix metalloproteinases in cerebrovascular disease. J. Cereb. Blood Flow Metab. 18, 1163–1172 (1998).
- Asahi M. et al. Effects of matrix metalloproteinase-9 gene knock-out on the proteolysis of blood-brain barrier and white matter components after cerebral ischemia. J. Neurosci. 21, 7724–7732 (2001).
- Lo E. H., Dalkara T. & Moskowitz M. A. Mechanisms, challenges and opportunities in stroke. Nat. Rev. Neurosci. 4, 399–415 (2003).
- Lo E. H., Broderick J. P. & Moskowitz M. A. tPA and proteolysis in the neurovascular unit. Stroke 35, 354–356 (2004).
- Pfefferkorn T. & Rosenberg G. A. Closure of the blood-brain barrier by matrix metalloproteinase inhibition reduces rtPA-mediated mortality in cerebral ischemia with delayed reperfusion. Stroke 34, 2025–2030 (2003).
- Lee S. R. & Lo E. H. Induction of caspase-mediated cell death by matrix metalloproteinases in cerebral endothelial cells after hypoxia-reoxygenation. J. Cereb. Blood Flow Metab. 24, 720–727 (2004).
- Gu Z. et al. A highly specific inhibitor of matrix metalloproteinase-9 rescues laminin from proteolysis and neurons from apoptosis in transient focal cerebral ischemia. J. Neurosci. 25, 6401–6408 (2005).
- Matsumoto S. et al. Expression and localization of matrix metalloproteinase-12 in the aorta of cholesterol-fed rabbits: relationship to lesion development. Am. J. Pathol. 153, 109–119 (1998).
- Chandler S., Cossins J., Lury J. & Wells G. Macrophage metalloelastase degrades matrix and myelin proteins and processes a tumour necrosis factor-alpha fusion protein. Biochem. Biophys. Res. Commun. 228, 421–429 (1996).
- Belaaouaj A. A., Li A., Wun T. C., Welgus H. G. & Shapiro S. D. Matrix metalloproteinases cleave tissue factor pathway inhibitor. Effects on coagulation. J. Biol. Chem. 275, 27123–27128 (2000).
- Dong Z., Kumar R., Yang X. & Fidler I. J. Macrophage-derived metalloelastase is responsible for the generation of angiostatin in Lewis lung carcinoma. Cell 88, 801–810 (1997).
- Cornelius L. A. et al. Matrix metalloproteinases generate angiostatin: effects on neovascularization. J. Immunol. 161, 6845–6852 (1998).
- Dwivedi A. & George S. MMP-12 is important for VSMC proliferation and migration: role of B-catenin signalling. Vascul Pharmacol 45, e129 (2006).
- Wells J. E. et al. Matrix metalloproteinase (MMP)-12 expression has a negative impact on sensorimotor function following intracerebral haemorrhage in mice. Eur. J. Neurosci. 21, 187–196 (2005).
- Svedin P., Hagberg H. & Mallard C. Expression of MMP-12 after neonatal hypoxic-ischemic brain injury in mice. Dev. Neurosci. 31, 427–436 (2009).
- Romanic A. M., White R. F., Arleth A. J., Ohlstein E. H. & Barone F. C. Matrix metalloproteinase expression increases after cerebral focal ischemia in rats: inhibition of matrix metalloproteinase-9 reduces infarct size. Stroke 29, 1020–1030 (1998).
- Montaner J. et al. Matrix metalloproteinase expression after human cardioembolic stroke: temporal profile and relation to neurological impairment. Stroke 32, 1759–1766 (2001).
- Svedin P., Hagberg H., Savman K., Zhu C. & Mallard C. Matrix metalloproteinase-9 gene knock-out protects the immature brain after cerebral hypoxia-ischemia. J. Neurosci. 27, 1511–1518 (2007).
- Hu Q. et al. Therapeutic application of gene silencing MMP-9 in a middle cerebral artery occlusion-induced focal ischemia rat model. Exp. Neurol. 216, 35–46 (2009).
- Bonoiu A. et al. MMP-9 gene silencing by a quantum dot-siRNA nanoplex delivery to maintain the integrity of the blood brain barrier. Brain Res. 1282, 142–155 (2009).
- Hu Q. et al. Lentivirus-mediated transfer of MMP-9 shRNA provides neuroprotection following focal ischemic brain injury in rats. Brain Res. 1367, 347–359 (2011).
- Mahajan S. D. et al. Suppression of MMP-9 expression in brain microvascular endothelial cells (BMVEC) using a gold nanorod (GNR)-siRNA nanoplex. Immunol. Invest 41, 337–355 (2012).
- Power C. et al. Intracerebral hemorrhage induces macrophage activation and matrix metalloproteinases. Ann. Neurol. 53, 731–742 (2003).
- Wasserman J. K., Zhu X. & Schlichter L. C. Evolution of the inflammatory response in the brain following intracerebral hemorrhage and effects of delayed minocycline treatment. Brain Res. 1180, 140–154 (2007).
- Gronski T. J. Jr et al. Hydrolysis of a broad spectrum of extracellular matrix proteins by human macrophage elastase. J. Biol. Chem. 272, 12189–12194 (1997).
- Liu Y. et al. Matrix metalloproteinase-12 contributes to neuroinflammation in the aged brain. Neurobiol. Aging 34, 1231–1239 (2013).
- Yong V. W., Power C., Forsyth P. & Edwards D. R. Metalloproteinases in biology and pathology of the nervous system. Nat. Rev. Neurosci. 2, 502–511 (2001).
- Churg A. et al. Acute cigarette smoke-induced connective tissue breakdown requires both neutrophils and macrophage metalloelastase in mice. Am. J. Respir. Cell Mol. Biol. 27, 368–374 (2002).
- Larsen P. H. & Yong V. W. The expression of matrix metalloproteinase-12 by oligodendrocytes regulates their maturation and morphological differentiation. J. Neurosci. 24, 7597–7603 (2004).
- Larsen P. H., DaSilva A. G., Conant K. & Yong V. W. Myelin formation during development of the CNS is delayed in matrix metalloproteinase-9 and -12 null mice. J. Neurosci. 26, 2207–2214 (2006).
- Vos C. M., van Haastert E. S., de Groot C. J., van der Valk P. & de Vries H. E. Matrix metalloproteinase-12 is expressed in phagocytotic macrophages in active multiple sclerosis lesions. J. Neuroimmunol. 138, 106–114 (2003).
- Ulrich R. et al. MMP-12, MMP-3, and TIMP-1 are markedly upregulated in chronic demyelinating theiler murine encephalomyelitis. J. Neuropathol. Exp. Neurol. 65, 783–793 (2006).
- Lanone S. et al. Overlapping and enzyme-specific contributions of matrix metalloproteinases-9 and -12 in IL-13-induced inflammation and remodeling. J. Clin. Invest 110, 463–474 (2002).
- Chelluboina B., Klopfenstein J. D., Gujrati M., Rao J. S. & Veeravalli K. K. Temporal regulation of apoptotic and anti-apoptotic molecules after middle cerebral artery occlusion followed by reperfusion. Mol. Neurobiol. 49, 50–65 (2014).
- Veeravalli K. K. et al. Human umbilical cord blood stem cells upregulate matrix metalloproteinase-2 in rats after spinal cord injury. Neurobiol. Dis. 36, 200–212 (2009).
Source: PubMed