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Dysregulation of miR-124 has been reported to be involved in the pathophysiology of depression.Chaihu-Shugan-San,a traditional Chinese medicine,has antidepressive activity;however,the underlying mechanisms remain unclear.In this study,to generate a rodent model of depression,rats were subjected to a combination of solitary confinement and chronic unpredictable mild stress for 28 days.Rats were intragastrically administered Chaihu-Shugan-San (2.835 mL/kg/d) for 4 weeks,once a day.Real-time reverse-transcription quantitative polymerase chain reaction,miRNA microarray,west blot assay and transmission electron microscopy demonstrated that ChaihuShugan-San downregulated miR-124 expression and upregulated the mRNA and protein levels of mitogen-activated protein kinase 14 (MAPK14) and glutamate receptor subunit 3 (Gria3).Chaihu-Shugan-San also promoted synapse formation in the hippocampus.The open field test,sucrose consumption test and forced swimming test were used to assess depression-like behavior.After intragastric administration of Chaihu-Shugan-San,sucrose consumption increased,while the depressive behaviors were substantially reduced.Together,these findings suggest that Chaihu-Shugan-San exerts an antidepressant-like effect by downregulating miR-124 expression and by releasing the inhibition of the MAPK 14 and Gria3 signaling pathways.