The Astrocytes Of Rat Cultured In Vitro And The Effect On The GFAP MRNA Expression After The Especially Intergenic Suppression

Objective:Through observation and analysis the change of the astrocytic morphous and the expression of GFAP after the astrocytes (AS) was cultured in vitro and transfected by different siRNA,we expect obtain significant gene suppressing effect and pay the way for the further correlative research.Methods:The cortical tissue of neonatal SD rat brain isolated was digested with 0.125% trypsin and made into single-cell suspension, which was cultured with DMEM/F12 medium containing 20% calf serum in the 50cm2 culture flask. The characteristics of AS in the different phase were detected by contrast microscopy and the cells was identified by using immunohistochemical method with antibody of rat GFAP labeled immunofluorescence. Then purified the cells by subculture. And the cells were randomly divided into six groups: three groups was experimental groups and each group was transfected with different siRNA of GFAP (siRNA1, siRNA2, siRNA3); one group was the positive group,one was the negative group, the other was the contrast group.Each group has eight specimen. Then detected the GFAP mRNA expression by using Western-blotting method and immunohistochemical method.Results: The purifed astrocytes were obtained and the cells were identified by using anti-rat GFAP body labeled with immunofluorescence. The experimental groups'fluorescence intensity decreased in different degree compared with the contrast group and negative group. The amount of astrocytes decreased and their cytodendrite shrinked after siRNA...