Abstract： Objective： To observe the clinical efficacy of Lingnan Tiaoshen Zhuyun acupuncture in treating premature ovarian insufficiency （POI） and its effect on serum sex hormone levels. Methods：A total of 62 POI patients in the First Affiliated Hospital of Guangzhou University of Chinese Medicine from March to November 2024 were selected and randomly divided into the observation group and the control group in a 1∶1 ratio using the random number table method，with 31 cases in each group. A total of 30 cases were included in each group after excluding the dropped out cases. The control group was treated with conventional acupuncture， while the observation group was treated with Lingnan Tiaoshen Zhuyun acupuncture. After three menstrual cycles of treatment， the clinical efficacy of the two groups was evaluated， as well as the changes in traditional Chinese medicine syndrome scores， Self-Rating Anxiety Scale （SAS） scores，Self-Rating Depression Scale （SDS） scores，serum follicle stimulating hormone （FSH）， luteinizing hormone （LH），FSH/LH ratio，and estradiol （E2） levels were evaluated in the two groups before and after treatment. Results： After treatment， the total effective rate was 96.67% （29/30） in the observation group， significantly higher than that of 80.00%（ 24/30） in the control group，the difference being significant（ P＜0.05）. After treatment，traditional Chinese medicine syndrome scores in both groups decreased compared to before treatment（ P＜0.05）. The difference （degree of improvement） in traditional Chinese medicine syndrome scores before and after treatment in the observation group was greater than that in the control group （P＜0.05）. After treatment， FSH and LH levels decreased （P＜0.05），while E2 levels increased （P＜0.05） in both groups，with no significant difference in the FSH/ LH ratio before and after treatment （P＞0.05）. The observation group showed lower FSH levels and FSH/LH ratio （P＜ 0.05） and greater improvements in FSH and E2 levels （P＜0.05） compared to the control group. After treatment，SAS and SDS scores decreased in both groups （P＜0.05）， with lower scores in the observation group （P＜0.05）. Conclusion：Lingnan Tiaoshen Zhuyun acupuncture improves clinical outcomes，regulates serum sex hormone levels， alleviates clinical symptoms，and reduces anxiety and depression in POI patients.