Abstract
The role of vascular endothelial growth factor (VEGF) in early brain injury (EBI) after subarachnoid hemorrhage (SAH) remains unclear. The aim of this study was to investigate effects of anti-VEGF therapy on EBI after SAH. C57BL/6 male mice underwent sham or filament perforation SAH modeling, and vehicle or two dosages (0.2 and 1 μg) of anti-VEGF antibody were randomly administrated by an intracerebroventricular injection. Neuroscore, brain water content, immunoglobulin G staining, and Western blotting were performed to evaluate EBI at 24–48 h. To confirm the role of VEGF, anti-VEGF receptor (VEGFR)-2 (a major receptor of VEGF) antibody was intracerebroventricularly administered and the effects on EBI were evaluated at 24 h. A higher dose, but not a lower dose, of anti-VEGF antibody significantly ameliorated post-SAH neurological impairments and brain edema at 24–48 h post-SAH. Post-SAH blood-brain barrier disruption was also inhibited by anti-VEGF antibody. The protective effects of anti-VEGF antibody were associated with the inhibition of post-SAH induction of VEGF, VEGFR-2, phosphorylated VEGFR-2, interleukin-1β and a matricellular protein tenascin-C (TNC). Anti-VEGFR-2 antibody also suppressed post-SAH neurological impairments and brain edema associated with VEGFR-2 inactivation and TNC downregulation. These findings demonstrated that VEGF causes post-SAH EBI via VEGFR-2 and TNC and that anti-VEGF therapy is effective for post-SAH EBI.
Similar content being viewed by others
Abbreviations
- ANOVA:
-
Analysis of variance
- BBB:
-
Blood-brain barrier
- CSF:
-
Cerebrospinal fluid
- EBI:
-
Early brain injury
- EGF:
-
Epidermal growth factor
- IL:
-
Interleukin
- MAPK:
-
Mitogen-activation protein kinase
- PBS:
-
Phosphate-buffered saline
- PDGF:
-
Platelet-derived growth factor
- p-VEGFR-2:
-
Phosphorylated vascular endothelial growth factor receptor-2
- SAH:
-
Subarachnoid hemorrhage
- TNC:
-
Tenascin-C
- VEGF:
-
Vascular endothelial growth factor
- VEGFR:
-
Vascular endothelial growth factor receptor
- ZO:
-
Zona occludens
References
Cahill J, Calvert JW, Zhang JH (2006) Mechanisms of early brain injury after subarachnoid hemorrhage. J Cereb Blood Flow Metab 26:1341–1353
Fujii M, Yan J, Rolland WB et al (2013) Early brain injury, an evolving frontier in subarachnoid hemorrhage research. Transl Stroke Res 4:432–446
Ferrara N, Gerber HP, LeCouter J (2003) The biology of VEGF and its receptors. Nat Med 9:669–676
Jiang S, Xia R, Jiang Y et al (2014) Vascular endothelial growth factors enhance the permeability of the mouse blood-brain barrier. PLoS One 9:e86407
Argaw AT, Asp L, Zhang J et al (2012) Astrocyte-derived VEGF-A drives blood-brain barrier disruption in CNS inflammatory disease. J Clin Invest 122:2454–2468
Kusaka G, Ishikawa M, Nanda A et al (2004) Signaling pathways for early brain injury after subarachnoid hemorrhage. J Cereb Blood Flow Metab 24:916–925
Tucker RP, Chiquet-Ehrismann R (2009) The regulation of tenascin expression by tissue microenvironments. Biochim Biophys Acta 1793:888–892
Suzuki H, Kanamaru K, Shiba M et al (2011) Cerebrospinal fluid tenascin-C in cerebral vasospasm after aneurysmal subarachnoid hemorrhage. J Neurosurg Anesthesiol 23:310–317
Shiba M, Fujimoto M, Imanaka-Yoshida K et al (2014) Tenascin-C causes neuronal apoptosis after subarachnoid hemorrhage in rats. Transl Stroke Res 5:238–247
Altay O, Suzuki H, Hasegawa Y et al (2012) Isoflurane attenuates blood-brain barrier disruption in ipsilateral hemisphere after subarachnoid hemorrhage in mice. Stroke 43:2513–2516
Chi OZ, Hunter C, Liu X et al (2007) Effects of anti-VEGF antibody on blood-brain barrier disruption in focal cerebral ischemia. Exp Neurol 204:283–287
Krum JM, Mani N, Rosenstein JM (2008) Roles of the endogenous VEGF receptors flt-1 and flk-1 in astroglial and vascular remodeling after brain injury. Exp Neurol 212:108–117
Suzuki H, Zhang JH (2012) Neurobehavioral assessments of subarachnoid hemorrhage. In: Chen J, Xu X-M, Xu ZC, Zhang JH (eds) Springer protocols handbooks. Animal models of acute neurological injuries II. Humana, New York, pp 435–440
Richmon JD, Fukuda K, Maida N et al (1998) Induction of heme oxygenase-1 after hyperosmotic opening of the blood-brain barrier. Brain Res 780:108–118
Suzuki H, Ayer R, Sugawara T et al (2010) Protective effects of recombinant osteopontin on early brain injury after subarachnoid hemorrhage in rats. Crit Care Med 38:612–618
Zacharia BE, Hickman ZL, Grobelny BT et al (2010) Epidemiology of aneurysmal subarachnoid hemorrhage. Neurosurg Clin N Am 21:221–233
Friedrich V, Flores R, Muller A et al (2010) Escape of intraluminal platelets into brain parenchyma after subarachnoid hemorrhage. Neuroscience 165:968–975
Scholler K, Trinkl A, Klopotowski M et al (2007) Characterization of microvascular basal lamina damage and blood-brain barrier dysfunction following subarachnoid hemorrhage in rats. Brain Res 1142:237–246
Takahashi H, Shibuya M (2005) The vascular endothelial growth factor (VEGF)/VEGF receptor system and its role under physiological and pathological conditions. Clin Sci 109:227–241
Ostrowski RP, Colohan AR, Zhang JH (2005) Mechanisms of hyperbaric oxygen-induced neuroprotection in a rat model of subarachnoid hemorrhage. J Cereb Blood Flow Metab 25:554–571
Suzuki H, Hasegawa Y, Kanamaru K et al (2010) Mechanisms of osteopontin-induced stabilization of blood-brain barrier disruption after subarachnoid hemorrhage in rats. Stroke 41:1783–1790
Yatsushige H, Ostrowski RP, Tsubokawa T et al (2007) Role of c-Jun N-terminal kinase in early brain injury after subarachnoid hemorrhage. J Neurosci Res 85:1436–1448
Zhang J, Xu X, Zhou D et al (2014) Possible role of Raf-1 kinase in the development of cerebral vasospasm and early brain injury after experimental subarachnoid hemorrhage in rats. Mol Neurobiol. doi:10.1007/s12035-014-8939-7
Shibuya M (2013) Vascular endothelial growth factor and its receptor system: physiological functions in angiogenesis and pathological roles in various diseases. J Biochem 153:13–19
Davis B, Tang J, Zhang L et al (2010) Role of vasodilator stimulated phosphoprotein in VEGF induced blood-brain barrier permeability in endothelial cell monolayers. Int J Dev Neurosci 28:423–428
Midwood KS, Hussenet T, Langlois B et al (2011) Advances in tenascin-C biology. Cell Mol Life Sci 68:3175–3199
Udalova IA, Ruhmann M, Thomson SJ et al (2011) Expression and immune function of tenascin-C. Crit Rev Immunol 31:115–145
Fujimoto M, Suzuki H, Shiba M et al (2013) Tenascin-C induces prolonged constriction of cerebral arteries in rats. Neurobiol Dis 55:104–109
Shiba M, Suzuki H, Fujimoto M et al (2012) Imatinib mesylate prevents cerebral vasospasm after subarachnoid hemorrhage via inhibiting tenascin-C expression in rats. Neurobiol Dis 46:172–179
Tanaka K, Hiraiwa N, Hashimoto H et al (2004) Tenascin-C regulates angiogenesis in tumor through the regulation of vascular endothelial growth factor expression. Int J Cancer 108:31–40
Fujimoto M, Shiba M, Kawakita F et al. (2015) Deficiency of tenascin-C attenuates blood-brain barrier disruption after experimental subarachnoid hemorrhage in mice. J Neurosurg. In press
Li W, Lu ZF, Man XY et al (2012) VEGF upregulates VEGF receptor-2 on human outer root sheath cells and stimulates proliferation through ERK pathway. Mol Biol Rep 39:8687–8694
Sozen T, Tsuchiyama R, Hasegawa Y et al (2009) Role of interleukin-1beta in early brain injury after subarachnoid hemorrhage in mice. Stroke 40:2519–2525
Chiquet-Ehrismann R, Chiquet M (2003) Tenascins: regulation and putative functions during pathological stress. J Pathol 200:488–499
Kuriyama N, Duarte S, Hamada T et al (2011) Tenascin-C: a novel mediator of hepatic ischemia and reperfusion injury. Hepatology 54:2125–2136
Narita Y (2013) Drug review: safety and efficacy of bevacizumab for glioblastoma and other brain tumors. Jpn J Clin Oncol 43:587–595
Stefanini FR, Badaro E, Falabella P et al (2014) Anti-VEGF for the management of diabetic macular edema. J Immunol Res 2014:632307
Acknowledgments
We thank Ms. Chiduru Yamamoto (Department of Neurosurgery, Mie University Graduate School of Medicine) for her technical assistance. This work was supported by a Grant-in-Aid for Scientific Research from Mie University Hospital Seed Grant Program 2014 to Dr. Suzuki, and Japan Society for the Promotion of Science to Dr. Fujimoto.
Conflict of Interest
The authors declare that they have no conflict of interest.
Research Involving Animals
All procedures were approved by the Animal Ethics Review Committee of Mie University, and were carried out according to the institution’s Guidelines for Animal Experiments.
Funding
This work was funded by a Grant-in-Aid for Scientific Research from Mie University Hospital Seed Grant Program 2014 to Dr. Suzuki, and Japan Society for the Promotion of Science to Dr. Fujimoto.
Author information
Authors and Affiliations
Corresponding author
Electronic Supplementary Material
Below is the link to the electronic supplementary material.
ESM 1
(PDF 5366 kb)
Rights and permissions
About this article
Cite this article
Liu, L., Fujimoto, M., Kawakita, F. et al. Anti-Vascular Endothelial Growth Factor Treatment Suppresses Early Brain Injury After Subarachnoid Hemorrhage in Mice. Mol Neurobiol 53, 4529–4538 (2016). https://doi.org/10.1007/s12035-015-9386-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12035-015-9386-9