Model preparation and evaluation
The model, acupuncture, and ZMTM groups received a daily subcutaneous injection of 4-VCD at 80 mg/kg for 15 consecutive days. The 200 g rats were given a volume of 0.2 ml (80 mg/ml), i.e., 0.001 ml/g, for 15 consecutive days to induce the perimenopausal model. After five days of treatment, rat vaginal smears were collected from each group for five consecutive days. The estrous cycle of rats was confirmed by microscopic evaluation of cells in vaginal smears. In all three groups, besides the standard control group, rat vaginal smears showed estrous symptoms, i.e., a significant amount of leukocytes with some epithelial cells and keratinocytes were present, which implies that the perimenopausal model was induced successfully. No mortality was seen during the induction of the perimenopausal phase.