To exploring the changes in vaginal microenvironment of patients with cervical lesions by high-risk human papillomavirus (HR-HPV) infection before and after treatment. Methods: 194 patients who had been diagnosed with HR-HPV infection and had undergone colposcopy biopsy in the second affiliated hospital of Zhengzhou university from September 2021 to December 2022 were selected in this study. The results of the cervical biopsy of these patients were LSIL or HSIL. And 194 patients had accepted the detections of vaginal microbiota and HPV before biopsy and in 6 months after treatment, and among them, there were 100 patients with LSIL in group A and 94 patients with HSIL in group B. The patients in group A were treated with interferon, and the patients in group B were treated with cervical conization. Another 188 patients with the negative results of HPV infection and thinprep cytologic test (TCT) during the same period were selected in control group. The changes and the correlations of the vaginal microecological indexes of the patients before and after treatment were analyzed. Results: Before treatment, the cleanliness degree, the white blood cell count, the dominant bacteria rate and the abnormal rate of Nugent score of the patients in group A were significantly higher than those of the patients in the control group (P<0.05). After treatment, the cleanliness degree, the white blood cell count, the dominant bacteria, the Nugent score and the abnormal rate of leukocyte esterase of the patients were significantly lower than those of the patients before treatment (P<0.05). Before treatment, the flora density, the flora diversity, the dominant bacteria, the Nugent score and the abnormal pH rate of the patients in group B were significantly higher than those of the patients in the control group (P<0.05). After treatment, the bacterial diversity, the dominant bacteria, the pH value and the abnormal rate of leukocyte esterase of the patients in group B were significantly lower than those of the patients before treatment (P<0.05). After treatment, there were 32 (32.0%) cases with HR-HPV turned negative in group A and there were 76 (80.9%) cases with HR-HPV turned negative in group B. After treatment, the dominant bacteria, the Nugent score, the pH value and the abnormal rate of hydrogen peroxide of the patients with HR-HPV positive were significantly higher than those of the patients with HR-HPV negative (P<0.05). Conclusion: The vaginal microbiota of the patients is closely related to the occurrence, development and conversion of their cervical lesions by HR-HPV infection.