title: Therapeutic application of gene silencing MMP-9 in a middle cerebral artery occlusion-induced focal ischemia rat model.
abstract: RNA interference appears to have a great potential not only as an in vitro target validation, but also as a novel therapeutic strategy based on the highly specific and efficient silencing of a target gene. We hypothesize that MMP-9 siRNA can be effective as an MMP-9 protein inhibitor in a rat focal ischemia model. Male Sprague-Dawley rats (156) were subjected to 2 h of middle cerebral artery occlusion (by using the suture insertion method) followed by 24 h of reperfusion. In the treatment group, 5 microl MMP-9 siRNA was administrated by intracerebroventricular injection within 60 min after 2 h of focal ischemia. The siRNA transfection was demonstrated by fluorescence conjugated siRNA. Treatment with MMP-9 siRNA produced a significant reduction in the cerebral infarction volume, brain water content, mortality rate and accompanying neurological deficits. The followings were recorded: Evan's blue and IgG extravasation were reduced; the expression of MMP-9 mRNA and protein were significantly silenced; and immunohistochemistry and Western blot analysis revealed that the expression of MMP-9 and VEGF were reduced while occludin and collagen-IV were up-regulated in brain tissues. Our findings provide evidence that a liposomal formulation of siRNA might be used in vivo to silence the MMP-9 gene and could potentially serve as an important therapeutic alternative in patients with cerebral ischemia.
authors.affiliation: Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Hu
authors.forename: Chunhua
authors.initials: C
authors.lastname: Chen
authors.forename: Junhao
authors.initials: J
authors.lastname: Yan
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Yang
authors.forename: Xianzhong
authors.initials: X
authors.lastname: Shi
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.forename: Jiliang
authors.initials: J
authors.lastname: Lei
authors.forename: Lei
authors.initials: L
authors.lastname: Yang
authors.forename: Ke
authors.initials: K
authors.lastname: Wang
authors.forename: Lin
authors.initials: L
authors.lastname: Chen
authors.forename: Hongyun
authors.initials: H
authors.lastname: Huang
authors.forename: Jingyan
authors.initials: J
authors.lastname: Han
authors.forename: John H
authors.initials: JH
authors.lastname: Zhang
authors.forename: Changman
authors.initials: C
authors.lastname: Zhou
mesh_terms: D000818:Animals; D001921:Brain / Q000098:blood supply / Q000201:enzymology / Q000503:physiopathology; D001929:Brain Edema / Q000201:enzymology / Q000235:genetics / Q000628:therapy; D020520:Brain Infarction / Q000201:enzymology / Q000235:genetics / Q000628:therapy; D024141:Collagen Type IV / Q000378:metabolism; D004195:Disease Models, Animal; D015536:Down-Regulation / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D020925:Hypoxia-Ischemia, Brain / Q000201:enzymology / Q000235:genetics / Q000628:therapy*; D020244:Infarction, Middle Cerebral Artery / Q000201:enzymology / Q000235:genetics / Q000628:therapy*; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D008565:Membrane Proteins / Q000378:metabolism; D062793:Occludin; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection / Q000379:methods; D015854:Up-Regulation / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism
chemical_list: D024141:Collagen Type IV; D061965:Matrix Metalloproteinase Inhibitors; D008565:Membrane Proteins; D062793:Occludin; C568987:Ocln protein, rat; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A; D020780:Matrix Metalloproteinase 9