Multiple effects of 2ME2 and D609 on the cortical expression of HIF-1alpha and apoptotic genes in a middle cerebral artery occlusion-induced focal ischemia rat model.
Despite 2-methoxyestradiol (2ME2) and tricyclodecan-9-yl-xanthogenate (D609) having multiple effects on cancer cells, mechanistically, both of them down-regulate hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF). We hypothesize HIF-1alpha plays an essential role in cerebral ischemia as a pro-apoptosis regulator; 2ME2 and D609 decrease the levels of HIF-1alpha and VEGF, that might contribute to protecting brain from ischemia injury. A total of 102 male Sprague-Dawley rats were split into five groups: sham, middle cerebral artery occlusion (MCAO), MCAO + dimethyl sulfoxide, MCAO + 2ME2, and MCAO + D609. 2ME2 and D609 were injected intraperitoneally 1 h after reperfusion. Rats were killed at 24 h and 7 days. At 24 h, 2ME2 and D609 reduce the levels of HIF-1alpha and VEGF (enzyme-linked immunosorbent assay), depress the expression of HIF-1alpha, VEGF, BCL2/adenovirus E1B 19 kDa interacting protein 3 (BNIP3) and cleaved caspase 3 (western blot and immunohistochemistry) in the brain infarct area. Double fluorescence labeling shows HIF-1alpha positive immunoreactive materials are co-localized with BNIP3 and terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling inside the nuclei of neurons. At 7 days, 2ME2 and D609 reduce the infarct volume (2,3,7-triphenyltetrazolium chloride) and blood-brain barrier extravasation, decrease the mortality and improve the neurological deficits. In conclusion, 2ME2 and D609 are powerful agents to protect brain from cerebral ischemic injury by inhibiting HIF-1alpha expression, attenuating the superfluous expression of VEGF to avoid blood-brain barrier disruption and suppressing neuronal apoptosis via BNIP3 pathway.
Chen
Chunhua
C
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, ChinaCenter of Tasly Microcirculation, Peking University Health Science Center, Beijing, ChinaDepartment of Neurosurgery, Louisiana State University Health Science Center in Shreveport, Louisiana, USA.
Hu
Qin
Q
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, ChinaCenter of Tasly Microcirculation, Peking University Health Science Center, Beijing, ChinaDepartment of Neurosurgery, Louisiana State University Health Science Center in Shreveport, Louisiana, USA.
Yan
Junhao
J
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, ChinaCenter of Tasly Microcirculation, Peking University Health Science Center, Beijing, ChinaDepartment of Neurosurgery, Louisiana State University Health Science Center in Shreveport, Louisiana, USA.
Lei
Jiliang
J
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, ChinaCenter of Tasly Microcirculation, Peking University Health Science Center, Beijing, ChinaDepartment of Neurosurgery, Louisiana State University Health Science Center in Shreveport, Louisiana, USA.
Qin
Lihua
L
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, ChinaCenter of Tasly Microcirculation, Peking University Health Science Center, Beijing, ChinaDepartment of Neurosurgery, Louisiana State University Health Science Center in Shreveport, Louisiana, USA.
Shi
Xianzhong
X
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, ChinaCenter of Tasly Microcirculation, Peking University Health Science Center, Beijing, ChinaDepartment of Neurosurgery, Louisiana State University Health Science Center in Shreveport, Louisiana, USA.
Luan
Liju
L
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, ChinaCenter of Tasly Microcirculation, Peking University Health Science Center, Beijing, ChinaDepartment of Neurosurgery, Louisiana State University Health Science Center in Shreveport, Louisiana, USA.
Yang
Lei
L
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, ChinaCenter of Tasly Microcirculation, Peking University Health Science Center, Beijing, ChinaDepartment of Neurosurgery, Louisiana State University Health Science Center in Shreveport, Louisiana, USA.
Wang
Ke
K
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, ChinaCenter of Tasly Microcirculation, Peking University Health Science Center, Beijing, ChinaDepartment of Neurosurgery, Louisiana State University Health Science Center in Shreveport, Louisiana, USA.
Han
Jingyan
J
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, ChinaCenter of Tasly Microcirculation, Peking University Health Science Center, Beijing, ChinaDepartment of Neurosurgery, Louisiana State University Health Science Center in Shreveport, Louisiana, USA.
Nanda
Anil
A
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, ChinaCenter of Tasly Microcirculation, Peking University Health Science Center, Beijing, ChinaDepartment of Neurosurgery, Louisiana State University Health Science Center in Shreveport, Louisiana, USA.
Zhou
Changman
C
Department of Anatomy and Embryology, Peking University Health Science Center, Beijing, ChinaCenter of Tasly Microcirculation, Peking University Health Science Center, Beijing, ChinaDepartment of Neurosurgery, Louisiana State University Health Science Center in Shreveport, Louisiana, USA.
D000077584:2-Methoxyestradiol; D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000502:physiology; D001929:Brain Edema / Q000188:drug therapy / Q000378:metabolism / Q000503:physiopathology; D020520:Brain Infarction / Q000188:drug therapy / Q000378:metabolism / Q000503:physiopathology; D002545:Brain Ischemia / Q000188:drug therapy* / Q000378:metabolism / Q000503:physiopathology; D001952:Bridged-Ring Compounds / Q000494:pharmacology*; D053148:Caspase 3 / Q000187:drug effects / Q000378:metabolism; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism; D004195:Disease Models, Animal; D015536:Down-Regulation / Q000187:drug effects / Q000502:physiology; D004958:Estradiol / Q000031:analogs & derivatives* / Q000494:pharmacology; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D020287:In Situ Nick-End Labeling; D020244:Infarction, Middle Cerebral Artery / Q000188:drug therapy / Q000378:metabolism / Q000503:physiopathology; D008297:Male; D008565:Membrane Proteins / Q000187:drug effects / Q000378:metabolism; D024101:Mitochondrial Proteins; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009636:Norbornanes; D011518:Proto-Oncogene Proteins / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013859:Thiocarbamates; D013871:Thiones / Q000494:pharmacology*; D016896:Treatment Outcome; D042461:Vascular Endothelial Growth Factor A / Q000187:drug effects / Q000378:metabolism
C505650:BNIP3 protein, rat; D001952:Bridged-Ring Compounds; C497444:Hif1a protein, rat; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D018696:Neuroprotective Agents; D009636:Norbornanes; D011518:Proto-Oncogene Proteins; D013859:Thiocarbamates; D013871:Thiones; D042461:Vascular Endothelial Growth Factor A; D004958:Estradiol; C046498:tricyclodecane-9-yl-xanthogenate; D000077584:2-Methoxyestradiol; D053148:Caspase 3