ABSTRACT Objective: This study aimed to investigate the effects of moxibustion on the expression of kisspeptin-related proteins and its association with the ALK/JNK/NLRP3 pyroptosis signaling pathway in rats with chemotherapy-induced Premature ovarian insufficiency (POI) induced by cyclophosphamide (Cy). We sought to evaluate the therapeutic potential of moxibustion and its protective effects against Cy-induced POI. Methods: A POI model was established. Moxibustion treatment was administered at acupoints SP6 and bilateral CV4. Following the intervention, we compared the morphological characteristics of ovarian and uterine tissues, as well as the number and morphological changes of ovarian follicles. Pathological examination of ovarian tissue slices was performed. Serum levels of FSH, LH, E2, AMH, IL-1β, and IL-18 were quantified using enzyme-linked immunosorbent assay. Differences in ER mRNA levels were assessed using quantitative reverse transcription q-PCR. Western blotting was conducted to detect the expression of pyroptosis-related proteins NLRP3 and GSDMD, as well as NLRP3 inflammatory activation-related proteins ALK and JNK. The expression of kisspeptin, P450Arom, and FSHR proteins was also analyzed. Estrous cycle changes were statistically evaluated, and the increment in body weight along with changes in ovarian and other visceral mass indices were calculated. Results: The POI group exhibited significantly elevated levels of FSH, LH, IL-1β, and IL-18 following Cy intervention, along with upregulation of ALK,JNK,NLRP3,GSDMD proteins. In comparison to the control group, the POI group showed a decrease in ovarian follicle count, E2, AMH, Moxibustion altered the expression of kisspeptin, P450Arom, and FSHR, as well as the aforementioned proteins, alleviating biochemical abnormalities and delaying the progression of POI. Significance: These findings support the hypothesis that moxibustion may improve Cy-induced POI by mitigating NLRP3 inflammatory activation and modulating the expression of proteins such as kisspeptin. Targeting kisspeptin and inhibiting NLRP3 inflammatory activation and granulosa cell pyroptosis may represent a novel therapeutic strategy for POI