A total of 142 premenopausal women with symptomatic adenomyosis underwent ultrasound (US)-guided percutaneous microwave ablation (PMWA) at the Chinese PLA General Hospital. This study aimed to evaluate changes in serum pituitary, gonadal hormone and cancer antigen 125 (CA125) levels after US-guided PMWA. Therefore, estradiol (E2), follicle-stimulating hormone (FSH), prolactin (PRL) and CA125 levels were evaluated before ablation and at 3, 6, 9 and 12 months after ablation. No significant differences were observed in the E2 and FSH levels pre-ablation and during follow-up (E2: p = 0.933, p = 0.987, p = 0.106, p = 0.936; FSH: p = 0.552, p = 0.295, p = 0.414, p = 0.760). The mean absolute values of serum CA125 and PRL were significantly decreased at 3, 6, 9 and 12 months after ablation (CA125: p < 0.001, p < 0.001, p < 0.001, p = 0.003; PRL: p < 0.001, p < 0.001, p < 0.001, p < 0.001). A significant correlation between changes in CA125 levels and uterine volume was found (p < 0.001). No evidence of a decline in ovarian function was observed after US-guided PMWA.