Early inhibition of HIF-1alpha with small interfering RNA reduces ischemic-reperfused brain injury in rats.
Hypoxia-inducible factor-1 (HIF-1) plays an essential role in cerebral ischemia as a proapoptotic factor. We hypothesized that HIF-1alpha siRNA can protect the brain from ischemic damage by inhibiting HIF-1alpha induced apoptotic pathway at the RNA level in a rat focal ischemic model. Results showed that treatment with HIF-1alpha siRNA reduced the infarct volume, decreased mortality, improved neurological deficits and reduced Evans blue extravasation. The expression of HIF-1alpha mRNA (Real-Time PCR) and protein were significantly silenced and the immunohistochemistry and Western blot revealed the suppression of HIF-1alpha, VEGF, p53 and Caspase-3. Double fluorescence labeling showed HIF-1alpha positive immunoreactive materials were partly colocalized with NeuN, p53 and Caspase-3 in the injured cerebral cortex. This study showed that HIF-1alpha siRNA may protect the ischemic-reperfused neurons in vivo via inhibition of HIF-1alpha, its downstream VEGF and other apoptotic-related proteins such as p53 and Caspase-3 and may have potentials for the early treatment of ischemic cerebral stroke.
Chen
Chunhua
C
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing 100083, China.
Hu
Qin
Q
Yan
Junhao
J
Yang
Xiaomei
X
Shi
Xianzhong
X
Lei
Jiliang
J
Chen
Lin
L
Huang
Hongyun
H
Han
Jingyan
J
Zhang
John H
JH
Zhou
Changman
C
D000818:Animals; D017209:Apoptosis; D001921:Brain / Q000473:pathology / Q000503:physiopathology; D053148:Caspase 3 / Q000378:metabolism; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism*; D020244:Infarction, Middle Cerebral Artery / Q000401:mortality / Q000473:pathology / Q000628:therapy*; D008297:Male; D009419:Nerve Tissue Proteins / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000401:mortality / Q000473:pathology / Q000628:therapy*; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
C497444:Hif1a protein, rat; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53; D042461:Vascular Endothelial Growth Factor A; C467498:vascular endothelial growth factor A, rat; D053148:Caspase 3