Effect Of Retrovirus Carrying Antisense Vimentin CDNA On The Expression Of Intermediate Filaments Of Astrocytes

Objectives:Reactive gliosis is a prominent result of many types of insult to the central nervous system(CNS) and leads to the formation of reactive gliosis and ultimately the occurrence of glial scar that impedes the regeneration of axons. To investigate the role of intermediate filament in the formation of reactive gliosis and glial scar, in vitro scratch-injury model was established with astrocytes. The astrocyte monolayer was infected with retrovirus carrying recombinant antisense vimentin cDNA. The effect of antisense vimentin cDNA on the expression of vimentin and glial fibrillary acidic protein(GFAP) was examined by using RT-PCR, Western blot and immunofluorescence assays. The model of spinal cord injury(SCI) was also established to evaluate the influence of antisense vimentin on the expression of vimentin and GFAP in vivo. We also examined the effect of chondrointinase ABC on the mobility of SCI rats, and expression of vimentin and GFAP.Materials and MethodsPartâ… : The astrocyte monolayer on the coverslips was scratched with a sterile plastic pipette tip to create in vitro scratch-injury model. After infected with retrovirus carrying antisense vimentin cDNA, the cell growth curves for astrocytes was determined, and the expression of vimentin and GFAP of injured astrocytes was examined by using RT-PCR, western blot and immunofluresence assays, respectively.Partâ…¡: The model of spinal cord injury(SCI) was established by hemisecting spinal cord at T10-T11 level in rats. Basso Beattie and Bresnahan locomotor rating score(BBB) of rats and the expression of vimentin and GFAP were determined. We also examined the effect of antisense vimentin cDNA and chondroitinase ABC on the expression of vimentin and GFAP of the injured spinal cord.Results...