Lentivirus-mediated transfer of MMP-9 shRNA provides neuroprotection following focal ischemic brain injury in rats

• Published in: Brain research 2011-01, Vol.1367 (7), p.347-359
• Main Authors: Hu, Qin, Chen, Chunhua, Khatibi, Nikan H, Li, Li, Yang, Lei, Wang, Ke, Han, Jingyan, Duan, Weiming, Zhang, John H, Zhou, Changman
• Format: Article
• Language: English
• Subjects: Analysis of Variance; Animals; Biological and medical sciences; blood-brain barrier; Blood-Brain Barrier - metabolism; Blood-Brain Barrier - physiopathology; Blood–brain barrier (BBB); brain; Brain edema; Cell Line, Transformed; Cells, Cultured; Cerebral circulation. Blood-brain barrier. Choroid plexus. Cerebrospinal fluid. Circumventricular organ. Meninges; collagen; Disease Models, Animal; Evans Blue; Fundamental and applied biological sciences. Psychology; gene expression; Gene Expression Regulation - genetics; Gene Transfer Techniques; infarction; Infarction, Middle Cerebral Artery - prevention & control; ischemia; Lentivirus; Lentivirus - genetics; Lentivirus - physiology; Male; Matrix Metalloproteinase 9 - genetics; Matrix Metalloproteinase 9 - metabolism; Medical sciences; metalloproteinases; Middle cerebral artery occlusion (MCAO); MMP-9; Neurology; neurons; neuroprotective effect; occludins; permeability; Rats; Rats, Sprague-Dawley; RNA, Messenger - metabolism; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; shRNA; Single-Blind Method; surgery; Tetrazolium Salts; Trauma Severity Indices; Vascular diseases and vascular malformations of the nervous system; Vertebrates: nervous system and sense organs; water content; Western blotting
• ISSN: 0006-8993; 1872-6240
• DOI: 10.1016/j.brainres.2010.10.002

Abstract Various studies on focal cerebral ischemic models have implicated the direct activation and expression of matrix metalloproteinases (MMPs), especially MMP-9, as a key orchestrator of blood–brain barrier (BBB) disruption. Moreover, studies have shown that MMP-9 siRNA can protect the BBB from ischemia/reperfusion injury. In the present study, we investigated the neuroprotective role of a lentivirus vector-mediated mmp-9shRNA following focal cerebral ischemia—specifically assessing whether LV-mmp9shRNA silencing of MMP-9 mRNA could ameliorate BBB disruption and in turn reduce vascular permeability, neuronal cell death, and neurobehavioral deficits. Treatment was given 2 weeks prior to surgery using a lentivirus-mediated vector. Surgery was conducted using the established middle cerebral artery occlusion (MCAO) model in rats, while outcomes were measured 24 h after injury. Our results demonstrated a significant reduction in brain infarction volume, brain water content, and neurobehavioral deficits following LV-mmp9shRNA treatment. Additionally, Evans blue and IgG extravasation were reduced, MMP-9 mRNA expression was silenced, and Western blot analysis revealed a decreased expression of MMP-9 and VEGF with an increased expression of occludin and collagen IV in brain tissues. This suggests that successful delivery of LV-mmp9shRNA may ameliorate ischemic brain injury by preserving structural integrity and improving functional outcome.