Background: Kisspeptin, a neuropeptide, plays a crucial role in stimulating the release of gonadotropin-releasing hormone (GnRH), which is essential for triggering the luteinizing hormone (LH) surge and subsequent ovulation. Notably, kisspeptin levels increase just prior to ovulation and the LH surge, underscoring its significance in regulating the ovulatory process throughout various stages of the menstrual cycle. Objective: To investigate how kisspeptin influences and interacts with other hormonal changes related to reproduction in both younger and older menstruating women. Materials and methods: This study involved 30 healthy women within two different age ranges: 20-25 years and 40-45 years. They were divided into two groups, with 15 women in each group. Blood samples were collected on the eighth, sixteenth, and twenty-fourth days of the menstrual cycle to compare various parameters, including Serum Kisspeptin, Follicle-Stimulating Hormone (FSH), Luteinizing Hormone (LH), Estradiol, Progesterone, and Prolactin hormones. The comparison was made within each age group on different days and also between the two age groups on corresponding days. Results and Conclusion: On the 16th day, there were notable increases (p≤ 0.05) in Kisspeptin, luteinizing hormone (LH), and prolactin levels compared to days 8 and 24 in both the first and second groups, albeit to different extents. Additionally, on the same days, the second group exhibited significant rises (p≤0.05) in Kisspeptin and prolactin when compared to the first group, although there was no significant increase in luteinizing hormone (LH). On the 8th day, there were notable increases (p≤0.05) in follicular stimulating hormone (FSH) and estradiol levels, and these changes differed between the first and second groups. Furthermore, on the same days, the second group experienced a significant rise (p≤0.05) in FSH compared to the first group, although estradiol levels did not show a significant decrease. On the 24th day, there was a significant increase (p≤0.05) in progesterone levels when compared to the 8th and 16th days, and this increase was observed in both groups. Additionally, when comparing the two groups on the same days, the second group showed a non-significant reduction in progesterone levels compared to the first group. Kisspeptin plays a crucial role in regulating the hypothalamic-pituitary-gonadal axis and collaborates with other reproductive hormones to facilitate successful ovulation in young menstruating women.