Objective:To construct and indentify rat glutamine synthetase(GS) RNA interference expression vector and detect GS on cell adhesion of astrocytes. Methods:The rat GS specific siRNA sequence was designed by online software,and GS eukaryotic expression vector,GS-EGFP were co-transfected into Hela-G cells by LipofectamineTM 2000 regent. GFP expression level was used to identify the interference efficiency of GS siRNA. The effective siRNA sequence was inserted into eukaryotic expression vector pRNAT H1.l/Neo,and the vector was transfected into astrocytes. The expression of GS in astrocytes was detected by immunocytochemistry. After trasfected with GS siRNA vector,the effect of GS on cell adhersion was detected by actin specific immunocytochemistry. Results:GS siRNA inhibited the expression of GS-EGFP in Hela-G cells. The inserted sequence of GS siRNA was confirmed by DNA sequencing. The GS siRNA expression vector can specifically inhibit the expression of GS in astrocytes. GS RNA interference leads to a significant decrease of cell adhesion. Conclusion:The construction of rat GS siRNA eukaryotic expression vector was successful. The initial data showed GS level in astrocytes impacted on cell adhesion. The results of this study lay the foundation for further investigating the role of GS in reactive astrocytes after central nervous system injury.