Objective To explore the regulatory effects and changing characteristics of the compound of Colla Corii Asini powder and honey on the antioxidant activity and intestinal flora of mice.Methods Seventy male BALB/c mice of SPF (Specific Pathogen Free, SPF) grade are randomly divided into 7 groups (10 mice in each group), namely the blank control group, the model control group, the Colla Corii Asini group (0.5 g/kg BW), the low/high-dose honey groups (1.0/2.0 g/kg BW, respectively), and the low/high-dose compound groups (0.5+1.0/0.5+2.0 g/kg BW, respectively). Except for the blank group, oxidative stress models are established for the remaining groups. After 30 days of continuous intervention, relevant indicators are detected.Results Compared with the Colla Corii Asini group and the honey group, the high-dose compound group significantly reduces the levels of lipid peroxidation products (MDA) and protein carbonylation (PCO), while simultaneously increasing the levels of superoxide dismutase (SOD) and reduced glutathione (GSH) (P<0.05), confirming that the compound has a synergistic antioxidant effect, and the high-dose group exhibits the best effect. The compound alleviates the body's inflammatory response by inhibiting the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β). The results of genomic sequencing of the microbial flora in the intestinal contents show that the effects of sole intake of Colla Corii Asini or honey on the flora were not significant, while the intake of the compound of Colla Corii Asini and honey could improve microecological disorders and imbalances, with the high-dose compound demonstrating a better effect. In the intestinal flora of mice that ingested the low-dose and high-dose compounds, there is an increase in the content of Bacteroidetes and a decrease in the content of Firmicutes.Conclusion The compound of Colla Corii Asini and honey effectively enhances the body's free radical scavenging ability through its synergistic antioxidant activity. Meanwhile, it regulates the balance of the flora, significantly reduces oxidative stress damage, and contributes to maintaining the homeostasis of the intestinal microenvironment.