Abstract: Objective：To investigate the role of racemic ketamine in regulating the expressions of glial glutamate transporter 1 (GLT-1), Na+-K+ pump, and growth associated protein-43 (GAP-43) in primary astrocytes, and to explore the possible mechanism of ketamine on astrocytes. Methods：The primary astrocytes cultured in vitro were established and treated with ketamine. MK-801 and AP-5 were used as the control drugs. The expression of GLT-1, Na+-K+ pump, and GAP-43 in primary astrocytes after treatment with different drugs was detected by Western blotting. The leakage rate of lactate dehydrogenase (LDH) was detected to determine the cytotoxicity of ketamine treatment from 30 min to 24 h. Results：Compared with the blank control group, the expression of GLT-1 in astrocytes treated with ketamine for 30 min and 2 h was increased (P<0.05), the expression of Na+-K+ pump in ketamine treatment group for 15 min and 30 min as well as MK-801 treatment group for 6 h was up-regulated (P<0.05), while the expression of GAP-43 in ketamine and MK-801 treatment groups for 6-24 h was significantly decreased (P<0.05). The leakage rate of LDH in primary astrocytes cultured in vitro was not significantly different from that in the blank control group after treatment with ketamine (100 μmol/L), MK-801 (1 μmol/L ), or AP-5 (50 μmol/L) for 24 h. Conclusions：Racemic ketamine can up-regulate GLT-1 and Na+-K+ pump expression possibly via a non-N-methyl-D-aspartic acid receptor pathway, and 100 μmol/L ketamine treatment for 24 h shows no cytotoxic effect on the primary astrocytes cultured in vitro.