Abstract: The aim of this work was to study the influence of long term usage of diazepam on neuroactive ligand-receptor interaction pathway.The total mRNA was extracted from the whole brain of mice after long-term usage of diazepam,then hybridized with Oligo microarray and screened by the analysis of gene expression profiles.The differentially expressed genes were screened by bioinformatics and the change of neuroactive ligand-receptor interaction pathway was analyzed through Kyoto Encyclopedia of Genes and Genomes（KEGG）.Compared with the control group,16 genes transcript level in neuroactive ligand-receptor interaction pathway were significant difference,and 9 genes mRNA expression was up-regulated and 7 genes were down-regulated.The influence of long-term usage of diazepam on neuroactive ligand-receptor interaction signaling pathway was significant,which may be closely related to the adverse reaction after long-term usage of diazepam.