Effect On Expression Of Connective Tissue Growth Factor In Astrocytes Of Rat Induced By Transforming Growth Factor-β1

Objectives Glial scar is the universal consequence of damage or pathological process of central nervous system(CNS). Astrocyte(AS) is major cell component of gliocyte to make up glial scar.and characterized by gliosis in response to various kinds of damages of central nervous system. Cellular model with brain injured was established by rat cerebral cortical astrocytes in vitro culture stimulated by TGF–β1,a stimulating factor.Then,the change of AS morpH was observed. The change of quantity of GFAP (cytoskeletal protein of AS )and CTGF(downstream factor mediated TGF-β1to promote fibrosis)were investigated from gene translational level and protein synthetic level. Generate mechanism of gliosis after CNS injured was investigate via those changes. Therefore, the targe of suppressing adverse effect of gliocyte in treating nervous system disease by nerve stemcellular transplantion may be find.Methods 1.Rat cerebral cortical astrocytes in vitro culture :was in accordance with McCarthy,s cultural method and improved.through repetat treatment per cells adhering by different speed and passage.After thrice passages,cells were accredited by immunofluorescence. 2.Cultured astrocytes were incubation in TGF-β1 of different density for 24 hours.Then , GFAP expression was examined by reverse transcription-polymerase chain reaction (RT-PCR)and Western- blotting analysis (WB) while the change of AS morpHous was observed by pHase-contrast microscopy. AS of Gliosis induced by TGF-β1 was observed from above. 3. Connective tissue growth factor (CTGF )expression in AS of gliosis by incubation in TGF-β1 of different density was examined by reverse transcription-polymerase chain reaction (RT-PCR) and Western- blotting analysis (WB) . Effect of CTGF induced by TGF-β1 was studied from point of transcription and translation.Results 1.GFAP of cultured As by treated and passaged for three times were detected by immunofluorescence and.cells of GFAP(+) were majority(>95%).2. The morpHous of AS incubated in TGF-β1 was changed , the expressions of GFAP mRNA and protein were increased,and the effect became more and more significant followed density increased(P<0.05). 3. The expressions of CTGF mRNA and protein of AS incubated in TGF-β1 were increased,and the effect became more and more significant followed density increased(P<0.05). Conclusion 1.The method of culturing rat cerebral cortical astrocytes in vitro adopted by the study can gain cells of high purity and these cells can be used for research.2. To stimulate SD rat cerebral cortical AS in vitro culture by TGF-β1 can induce gliosis in As. 3. TGF-β1 can up-regulate CTGF expression in astrocytes of gliosis and the effect became more and more significant followed density increased. CTGF would be investigated further as the targe of suppressing adverse effect of gliocyte to treat nervous system disease .