Although it is known that high fructose intake causes salt-sensitive hypertension, the underlying mechanism remains unclear. The aim of this study was to determine whether chronic intake of high fructose coupled with salt (HFS) might alter the structure of the gut microbiota, which contributes to elevated blood pressure. For 8 wk, Sprague-Dawley rats were given 20% fructose in drinking water and 4% sodium chloride in their diet to induce hypertension. A non-absorbable antibiotic vancomycin was used to modify gut microbiota. The 16 S rRNA sequencing for fecal samples was assessed and blood pressure was recorded. Enzyme-linked immunosorbent assay and quantitative polymerase chain reaction were used to examine the renin-angiotensin system in serum, urine, and the kidney. Compared with the control group, HFS feeding resulted in gut dysbiosis by altering the diversity and richness of gut microbiota and decreased the ratio of Firmicutes to Bacteroidetes. Vancomycin reshaped dramatically the HFS-induced dysbiosis. And vancomycin (van) attenuated HFS-increased blood pressure (HFS: 121.3 ± 2.8 mm Hg; HFS-van: 111.1 ± 1.7 mm Hg) and heart rate (HFS: 360.5 ± 9.0 bpm; HFS-van: 318.7 ± 5.6 bpm) as well as the content of angiotensinogen, renin, and angiotensin II in the urine and the angiotensinogen mRNA level in renal cortical tissues. However, HFS-increased triacylglycerol, renin, and angiotensin II in serum were not decreased by vancomycin. The present results demonstrated that gut dysbiosis develops after chronic fructose plus salt intake and contributes to the increase of blood pressure and the activation of the intrarenal renin-angiotensin system. Therefore, targeting gut microbiota provides a helpful therapy method to improve HFS-induced hypertension.