Serum miRNA expression profile in responders to acupuncture treatment for diminished ovarian reserve: a study protocol for a nested case-control study

Objective Diminished ovarian reserve (DOR) leads to subfertility, infertility, and premature menopause, significantly compromising women’s reproductive health. Although clinical evidence supports the effectiveness of acupuncture in improving ovarian reserve, treatment responses vary considerably among DOR patients, and the factors associated with favorable outcomes remain unclear. Serum microRNAs may serve as potential biomarkers for distinguishing acupuncture responders, warranting further investigation. Design and methods This prospective nested case-control study will enroll women with DOR who undergo acupuncture treatment. Treatment response will be evaluated based on changes in anti-Müllerian hormone (AMH) levels, antral follicle count (AFC), serum follicle stimulating hormone (FSH) levels and clinical pregnancy. According to their response, participants demonstrating greater improvement will be assigned to the experimental group (responders), while non-responders will be matched in a 1:1 ratio to form the control group. Serum microRNA profiles from both groups will be analyzed using high-throughput sequencing to identify differentially expressed microRNAs. Candidate microRNAs will subsequently be validated via real-time quantitative PCR (RT-qPCR), and receiver operating characteristic (ROC) curve analysis will be performed to evaluate their predictive value for acupuncture treatment response. Discussion This is the first clinical study to characterize the serum miRNA expression profiles of DOR patients who benefit from acupuncture. The findings are expected to provide evidence supporting the precise identification of acupuncture-responsive individuals within this patient population. Study protocol registration https://itmctr.ccebtcm.org.cn/mgt/project/view/-4614215526207679163 , identifier ITMCTR2025000004.