title: Protection of granulocyte-colony stimulating factor to hemorrhagic brain injuries and its involved mechanisms: effects of vascular endothelial growth factor and aquaporin-4.
abstract: Granulocyte-colony stimulating factor (G-CSF) has protective effects on many neurological diseases. Here, we aimed to test G-CSF's effects on perihematomal tissue injuries following intracerebral hemorrhage (ICH) and examine whether the effects were functionally dependent on vascular endothelial growth factor (VEGF) and aquaporin-4 (AQP4). We detected the expression of perihematomal VEGF, VEGF receptors (VEGFRs) and AQP4 at 1, 3 and 7days after ICH. Also, we examined the effects of G-CSF on tissue injuries by ICH in wild type mice, and tested whether such effects were VEGF and AQP4 dependent by using VEGFR inhibitor - SU5416 and AQP4 knock-out (AQP4(-/-)) mice. Furthermore, we assessed the related signal transduction pathways via astrocyte cultures. We found G-CSF highly increased perihematomal VEGF, VEGFR-2 and AQP4. Importantly, G-CSF led to neurological functional improvement in both types of mice by associating with reduction of brain edema, blood-brain barrier (BBB) permeability and neuronal death and apoptosis and statistical analysis suggested AQP4 was required for these effects. Besides, except BBB leakage alleviation, the above effects were attenuated but not counteracted by SU5416, suggesting involvement of VEGF. G-CSF up-regulated phosphorylation of extracellular signal-regulated kinase (ERK) and signal transducer and activator of transcription 3 (STAT3) as well as VEGF and AQP4 proteins in cultured astrocytes. The latter was inhibited by ERK and STAT3 inhibitors respectively. Our data suggest the protective effects of G-CSF on perihematomal tissue injuries after ICH are highly associated with the increased levels of VEGF and AQP4, possibly act through C-Jun amino-terminal kinase and ERK pathways respectively.
authors.affiliation: Department of Neurology, Huashan Hospital, Fudan University, No.12 Middle Wulumuqi Road, Shanghai 200040, PR China.
authors.forename: H
authors.initials: H
authors.lastname: Chu
authors.affiliation: Department of Neurology, Huashan Hospital, Fudan University, No.12 Middle Wulumuqi Road, Shanghai 200040, PR China.
authors.forename: Y
authors.initials: Y
authors.lastname: Tang
authors.affiliation: Department of Neurology, Huashan Hospital, State Key Laboratory of Medical Neurobiology, Fudan University, No.12 Middle Wulumuqi Road, Shanghai 200040, PR China. Electronic address: qiang_dong163@163.com.
authors.forename: Q
authors.initials: Q
authors.lastname: Dong
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D051401:Aquaporin 4 / Q000378:metabolism*; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism; D001929:Brain Edema / Q000150:complications / Q000188:drug therapy; D003342:Corpus Striatum / Q000187:drug effects / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor / Q000627:therapeutic use*; D006801:Humans; D020300:Intracranial Hemorrhages / Q000150:complications / Q000188:drug therapy* / Q000378:metabolism; D008297:Male; D051379:Mice; D018696:Neuroprotective Agents / Q000627:therapeutic use*; D040262:Receptors, Vascular Endothelial Growth Factor / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: C496031:Aqp4 protein, mouse; D051401:Aquaporin 4; D018696:Neuroprotective Agents; D042461:Vascular Endothelial Growth Factor A; D016179:Granulocyte Colony-Stimulating Factor; D040262:Receptors, Vascular Endothelial Growth Factor
keywords: 4′,6-diamidino-2-phenylindole; ANOVA; AQP4; BBB; C-Jun amino-terminal kinase; DAPI; DMEM; Dulbecco’s Modified Eagle Medium; EB; EDTA; ELISA; ERK; Evans Blue; Flk-1; Flt-1; G-CSF; ICH; JNK; MAPK; PI3K; PMSF; STAT; TUNEL; VEGF; analysis of variance; aquaporin-4; blood–brain barrier; enzyme-linked immunosorbent assay; ethylenediaminetetraacetic acid; extracellular signal-regulated kinase; fams-like tyrosine kinase-1; fetal liver kinase-1; granulocyte-colony stimulating factor; intracerebral hemorrhage; mitogen-activated protein kinase; phenylmethanesulfonyl fluoride; phosphatidylinositol 3′-kinase; recombinant human G-CSF; rhG-CSF; signal transducer and activator of transcription; terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling; vascular endothelial growth factor