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

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.

• Authors: Qin Hu, Chunhua Chen, Nikan H Khatibi, Li Li, Lei Yang, Ke Wang, Jingyan Han, Weiming Duan, John H Zhang, Changman Zhou
• Journal: Brain research (Brain Res) Vol. 1367 Pg. 347-59 (Jan 07 2011) ISSN: 1872-6240 [Electronic] Netherlands
• PMID: 20950592 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2010 Elsevier B.V. All rights reserved.

Chemical References

• RNA, Messenger
• RNA, Small Interfering
• Tetrazolium Salts
• Evans Blue
• triphenyltetrazolium
• Matrix Metalloproteinase 9

Topics

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