Objective To investigate the effects of SOX6 gene interference on apoptosis, mitochondrial membrane potential, and the PI3K/ AKT signaling pathway of astrocytes (AS) induced by H₂O₂ . Methods AS cells were isolated from the cerebral cortex of neonatal rats and divided into the following groups: negative control group (NC group): AS transfected with siRNA without interference; H₂O₂ group: AS cells treated with 20 mol/LH₂O₂ for 24 h; H₂O₂ +NC group: non-interfering siRNA transfected into AS cells for 24 h, then treatment of cells with 20μmol /L H₂O₂ for 24 h;and H₂O₂+si-SOX6 group: SOX6-specific siRNA transfected into AS cells for 24 h, followed by treatment of the cells with 20 μmol / L H₂O₂ for 24 h. The cytotoxicity, apoptosis rate, and mitochondrial membrane potential were determined by LDH kit, flow cytometry, and JC-1 probe. The expression of SOX6, Cyt. C, caspase 3, caspase9, Bcl-2, Bax, and p-AKT protein was determined by Western blotting. Results H₂O₂ significantly upregulated the expression of SOX6, Cyt. C, caspase3, caspase9, and Bax; downregulated the expression of Bcl-2 and p-AKT; increased cytotoxicity; and induced apoptosis. Interfering with SOX6 gene expression significantly reduced the effects of H₂O₂ on SOX6, Cyt.C, caspase3, caspase9, Bcl-2, Bax, and p-AKT expression, cytotoxicity, and apoptosis. Conclusions Interference with SOX6 gene expression can protect against AS injury induced by H₂O₂ through the anti-apoptotic pathway, which is related to the mitochondrial pathway and PI3K/ AKT signaling activation.