Upregulation of vascular endothelial growth factor is associated with radiation-induced blood-spinal cord barrier breakdown.

Abstract

The pathogenesis of radiation-induced injury to the central nervous system (CNS) remains unclear. Dysfunction of the blood-brain barrier (BBB) is associated with radiation-induced white matter lesions. The aim of this study was to determine if vascular endothelial growth factor (VEGF) is implicated in radiation-induced BBB disruption. Adult rats were irradiated with a single dose of 8 or 22 Gy to the spinal cord from C2 to T2. At various times up to 20 weeks following irradiation, blood-spinal cord barrier (BSCB) permeability was assessed using immunohistochemistry with anti-albumin antibody. Cell proliferation was assessed using bromodeoxyuridine (BrdU), and endothelial cell identity was assessed morphologically and using immunostaining for factor VIII-related antigen. Expression of VEGF protein and message was assessed using immunohistochemistry and in situ hybridization respectively. In the unirradiated rat spinal cord, there was no evidence of albumin immunoreactivity and little evidence of VEGF expression. After a dose of 22 Gy, focal albumin staining in white matter was observed at 16 weeks. Diffuse staining was seen at 20 weeks and was associated with necrosis and demyelination in white matter. This was associated with a significant increase in white matter glial cells that showed immunoreactivity and in situ hybridization signal for VEGE VEGF expressing cells showed dual immunoreactivity for glial fibrillary acidic protein. No increase in VEGF positive cells was observed in gray matter after 22 Gy. After a dose of 8 Gy, there was no increase in VEGF expression or albumin immunostaining in either white or gray matter. Microvessel endothelial cell density showed a trend towards a decrease with time after 22 Gy as compared with 8 Gy or unirradiated controls. BrdU immunostaining provided no evidence for endothelial cell proliferation in control or in the irradiated spinal cord. It is concluded that radiation-induced BSCB dysfunction is associated with upregulation of VEGF in astrocytes without associated endothelial proliferation. The temporal and spatial association of VEGF upregulation with the white matter lesions suggests a role of VEGF in radiation-induced late CNS injury.