Aim To analyze the changes of the intestinal flora structure of two kidney one clip (2K1C) renovascular hypertension (RVH) rats, and to provide a new idea for the prevention and treatment of RVH. Methods 8 week old male Sprague-Dawley rats were randomly divided into 2K1C model group (n＝8) and sham operation group (n＝8), and RVH model was prepared by 2K1C surgery. The systolic pressure of tail artery was measured by non-invasive tail cuff method. At 4th week after the operation, intestinal feces were collected and the bacteria DNA was extracted from the feces. The diversity of intestinal flora in 2K1C rats was analyzed by PCR-denaturing gradient gel electrophoresis. The dominant bacteria (bifidobacterium and lactobacillus) in the intestinal tract were quantitatively detectd by real-time fluorescence quantitative PCR. Results Compared with the sham operation group, systolic pressure of tail artery in 2K1C model group was significantly higher than that in sham operation group (164.77 mmHg vs 122.08 mmHg, P＜0.05). However, there was no significant change in the systolic pressure of tail artery in sham operation group compared with the previous model (122.08 mmHg vs 120.13 mmHg, P＞0.05). There was a significant difference in intestinal flora diversity between 2K1C model group and sham operation group (P＜0.05), and intestinal flora diversity was lower in 2K1C model group. The contents of bifidobacterium and lactobacillus in 2K1C model group were significantly lower than those in sham operation group (2.335×10－3 ng/g vs 1.805×10－2 ng/g, 0.048 ng/g vs 0.139 ng/g, P＜0.05). Conclusion The diversity of intestinal flora in 2K1C RVH rats is significantly decreased, and the contents of bifidobacterium and lactobacillus are significantly changed, suggesting that it may be involved in the occurrence and development of hypertension in 2K1C rats.