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

Qin Hu, Chunhua Chen, Nikan H. Khatibi, Li Li, Lei Yang, Ke Wang, Jingyan Han, Weiming Duan, John H. Zhang, Changman Zhou

Research output: Contribution to journalArticlepeer-review

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. © 2010 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)347-359
Number of pages13
JournalBrain Research
Volume1367
DOIs
StatePublished - Jan 7 2011

ASJC Scopus Subject Areas

  • General Neuroscience
  • Molecular Biology
  • Clinical Neurology
  • Developmental Biology

Keywords

  • Blood-brain barrier (BBB)
  • Brain edema
  • Lentivirus
  • MMP-9
  • Middle cerebral artery occlusion (MCAO)
  • shRNA
  • Single-Blind Method
  • RNA, Small Interfering/genetics
  • Male
  • Tetrazolium Salts
  • Matrix Metalloproteinase 9/genetics
  • Gene Expression Regulation/genetics
  • Disease Models, Animal
  • Gene Transfer Techniques
  • Cells, Cultured
  • Rats
  • Blood-Brain Barrier/metabolism
  • Trauma Severity Indices
  • Rats, Sprague-Dawley
  • RNA, Messenger/metabolism
  • Infarction, Middle Cerebral Artery/prevention & control
  • Animals
  • Evans Blue
  • Analysis of Variance
  • Cell Line, Transformed
  • Lentivirus/genetics

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