Objective To investigate the effects of five-element music on depressive behaviors and intestinal flora in offspring of fear-stress rats during pregnancy. Methods Thirty-six 0. 5-day pregnant Wistar rats were divided randomly into a control group, fear-stress group, and five-element music group, and the subsequent litters continued the maternal grouping. Pregnant rats in the fear-stress and five-element music groups were modeled on days 1~19 of gestation using the bystander electric method, while the five-element music group was also exposed to Feather Tune five-element music intervention. The fear behavior and serum glucocorticoid ( GC) levels in pregnant rats were assessed on day 20 of pregnancy by open field test and enzyme-linked immunosorbent assay, respectively, to evaluate the effectiveness of the model. Depression in 3-week-old offspring was evaluated by open field, tail suspension, and sucrose preference tests. Norepinephrine(NE), dopamine (DA), and 5-hydroxytryptamine ( 5-HT) levels in the hippocampus of the offspring were measured by high-performance liquid chromatography. Changes in the intestinal flora of the offspring were analyzed by 16S rRNA sequencing. Expression levels of the proximal colonic proteins claudin1, occludin, and ZO1 in the offspring were measured by Western blot assay. Results Pregnant rats in the fear-stress group stayed longer in the area around the open field (P<0. 05), had fewer entries into the central area (P<0. 001), and had higher serum GC levels compared with those in the blank group (P<0. 001). In contrast, fiveelement music exposure reversed these behavioral changes (P<0. 05) and serum GC levels (P<0. 001) in the fearstress group. Offspring in the fear-stress group exhibited decreased open-field crossing frequency (P<0. 01), reduced sucrose preference index, and longer immobility time in the tail suspension test (P<0. 05), compared with those in the blank group, together with significantly decreased NE, DA, and 5-HT levels in the hippocampus (P<0. 05).Rats in the fear-stress group showed decreased diversity of the intestinal flora (P<0. 01) and significant alterations in flora structure, including higher abundance of Proteobacteria, Enterobacteriaceae, Enterococcus, and Escherichia (P<0. 05), and lower abundance of Spirochaetes, Spirochaetaceae, Lachnospiraceae, Ruminococcaceae, Treponema, Prevotella, Coprococcus, Allobaculum, Ruminococcus, and Dorea ( P<0. 05 ). The proximal colonic proteins Claudin1, Occludin, and ZO1 were significantly downregulated ( P<0. 05). The open-field crossing frequency,sucrose preference index, and duration of tail suspension immobilization were improved in the five-element music group compared with those in the fear-stress group (P<0. 05), while DA and 5-HT levels were significantly restored in the hippocampus (P<0. 05), species diversity of the intestinal flora increased (P<0. 01) and changes in the abundance of the flora were reversed, and the proximal colonic proteins Occludin and ZO1 expression were significantly upregulated (P<0. 05). Conclusions Five-element music intervention during pregnancy can ameliorate fear-stress-induced depression behaviors and intestinal flora disorders in the offspring.