Objective To explore the effect of Jianpi Yifei prescription (JPYF) on the expression of glutamatetransporter 2 (EAAT2) and the possible mechanism behind this. Methods Neonatal Sprague-Dawley rats within 24 h ofbirth were used and and the astrocytes of cerebral cortex were isolated under aseptic conditions, then purified, and culturedfor later experiments. The appropriate cell concentration was adjusted in plates for five groups: A, glutamic acid group (250μmoL/ L), B-D, JPYF prescription (final concentrations 0. 25, 0. 5, and 1 g/ kg) + glutamic acid group; and E, normalgroup, After the B-D groups had been treated with JPYF prescription (final concentrations of 0. 25, 0. 5, and 1 g/ kg) for24 h, glutamate at 250 μmol/ L was applied to the A-D groups for 4 h to induce an in vitro primary astrocyte injury model.Cell viability was determined by MTT assay. Determination of glutamic acid concentration in culture medium was performedby ultraviolet colorimetry. The expression of glutamate transporter protein in astrocytes was detected by immunofluorescenceand western blotting. The injury of spinal cord neurons was detected by Immunofluorescence. Results JPYF Fangsignificantly protected the astrocytes ( P < 0. 01), improved the expression of glutamate transporters ( P < 0. 05, P < 0. 01),reduced glutamate concentrations ( P < 0. 05, P < 0. 01), protectd neurons ( P < 0. 05, P < 0. 01), and showed a certaindose-related effect within the range of 0. 25-1 g.L^-1. Conclusions Within a certain dose range, JPYF Fang can reducethe damage of glutamate to neuronal cells, the mechanism of which may involve improving the expression of glutamate transporter and decreasing the concentration of glutamate in extracellular fluid, so as to protect neurons.