[Objectives]  To investigate whether bee pollen regulates the gut flora of hyperlipidemic rats (Rattus norvegicus). [Methods]  Hyperlipidemic rats were given three doses of bee pollen (80, 400 and 800 mg/kg) over 60 days, after which the structure and diversity of their gut bacteria was determined using 16S rDNA high-throughput sequencing. [Results]  The alpha diversity index (Shannon, Simpson) and richness index (Chao, Ace) indicate that the diversity of the rats’ gut bacteria declined, and that this was not reversed by bee-pollen gavage. The Firmicutes, Bacteroides and Verrucomicrobia accounted for more than 95.8% of all bacteria found in all three treatment groups. The abundance of Tenericutes in the high fat model group, and in the low and medium dose bee pollen groups, decreased significantly (P < 0.05), compared to the control group. At the genus level, the abundance of Bacteroides, Coprococcus and Paraprevotella decreased significantly in all three pollen- treatment groups compared to the control group (P < 0.05). Bacteroides abundance in the medium pollen-dose group increased significantly (P < 0.05) and the abundance of Akkermansia in all three pollen-treatment groups increased (P > 0.05) compared to the high-fat model group. [Conclusion]  The gut bacteria of hyperlipidemic rats can be altered by bee pollen.