Aim:Ketamine is an anesthetic adjunct widely applied in the clinic that can induce hippocampal neurodegeneration in the brain. MicroRNAs (miRNAs) have been shown to be related to the regulation of ketamine-mediated neurotoxicity. This investigation aimed to determine the function of miR-384-5p in ketamine-induced neurotoxicity.Material and Methods:Neonatal hippocampal neurons were isolated from rats and treated with varying doses of ketamine. RT–qPCR was utilized to measure the miR-384-5p level in ketamine-treated neurons. Neuronal viability was evaluated by MTT assay. TUNEL staining and flow cytometry were applied to measure neuronal apoptosis. H2-DCFDA staining was utilized to detect the intracellular ROS level. Protein levels were measured using Western blotting. A luciferase reporter experiment was used in HEK293T cells to verify the interaction of miR-384-5p with GABRB1.Results:Ketamine induced neurotoxicity and miR-384-5p upregulation in hippocampal neurons. MiR-384-5p downregulation mitigated ketamine-induced neurotoxicity by restraining apoptosis and ROS activity in neurons. GABRB1 was demonstrated to be targeted by miR-384-5p. GABRB1 depletion worsened ketamine-induced neurotoxicity. Moreover, GABRB1 depletion lessened the protective effect of miR-384-5p inhibition against ketamine-mediated neurotoxicity.Conclusion: MiR-384-5p regulates ketamine-induced neurotoxicity in hippocampal neurons by targeting GABRB1.