Therapeutic application of gene silencing MMP-9 in a middle cerebral artery occlusion-induced focal ischemia rat model.
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.
Hu
Qin
Q
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, China.
Chen
Chunhua
C
Yan
Junhao
J
Yang
Xiaomei
X
Shi
Xianzhong
X
Zhao
Jing
J
Lei
Jiliang
J
Yang
Lei
L
Wang
Ke
K
Chen
Lin
L
Huang
Hongyun
H
Han
Jingyan
J
Zhang
John H
JH
Zhou
Changman
C
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
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