【Abstract】Objective To study the effect of active ingredients of Xiongmatang on intestinal microbes in migraine rats. Methods 120 rats underwent dural cannulation surgery, observing the general physical signs of rats, 90 rats with better living conditions were selected and injected with inflammation soup 6 times within 12 days to stimulate the dura mater to replicate the migraine model, and were randomly divided into 9 groups: Model group, CGRP inhibitor group, Flunarizine positive group, the active ingredients of Xiongmatang(Low-dose group of n-butanol extract of Xiongmatang, High-dose group of n-butanol extract of Xiongmatang, Low-dose group of ethyl acetate extract of Xiongmatang, High-dose group of ethyl acetate extract of Xiongmatang, Low-dose group of n-butanol + ethyl acetate extract of Xiongmatang, High-dose group of n-butanol + ethyl acetate extract of Xiongmatang), 10 each group. Another 10 rats were injected with 0.9% sodium chloride solution in the same way as Control. After successful modeling, the blank group and the model group were given Pure water by gavage, and the rats in the CGRP inhibitor group were given administration by tail vein, once a week, for a total of 5 times; the other groups were given corresponding drugs, once a day, for a total of 4 weeks (28 d). At the end of the drug administration period, behavioral assessments were conducted using sucrose preference test, tail suspension test, novel object recognition test, and Morris water-maze test. Abdominal aortic blood was collected for ELISA quantification of circulating CGRP and NO levels. Intestinal contents were taken, and the changes in the gut microbiota of rats were analyzed by 16S rDNA high-throughput sequencing technology. Results Compared with the control group, model group rats exhibited significant depressive-like behavioral changes and cognitive impairments. Meanwhile, the contents of CGRP and NO in serum were significantly increased . Gut microbiota analysis revealed marked alterations in microbial community structure, at the phylum level, the community abundance of Firmicutes, Actinobacteriota, Unclassified and Campylobacterota decreased significantly. The community abundance of Bacteroidota, Verrucomicrobiota, Patescibacteria and Cyanobacteria increased significantly . At the genus level, community abundances such as Muribaculaceae_unclassified and Lactobacillus were significantly decreased . The community abundances of Ruminococcus, Clostridia_UCG-014_unclassified, Akkermansia, Firmicutes_unclassified and Monoglobus were significantly increased. Compared with the model group, the CGRP inhibitor group, flunarizine positive group, the active ingredients of Xiongmatang dose-dependently reversed the depressive-like behavioral changes and cognitive impairments observed in migraine rats, and significantly lowered serum levels of CGRP and NO. The active ingredients of Xiongmatang treatment also modulated intestinal microbial diversity: α-diversity assays showed increased richness and evenness, asβ-diversity revealed that the microbial community structure of XMT-treated rats closely resembled that of the blank group. At both phylum and genus levels, the alterations in microbiota composition exhibited trends opposite to those seen in the model group. Conclusions The active ingredients of Xiongmatang may prevent and treat migraine in rats by regulating the gut microbiota.