早发性卵巢功能不全患者血清microRNA表达谱对针刺疗效的预测价值研究

Premature ovarian insufficiency (POI) is an important cause for infertility. It severely affects women’s reproductive health. In modern medicine, there is still no effective methods on improving ovarian function. Researches showed that acupuncture can improve the ovarian function in women with POI, and increase their chances of receiving assisted reproductive technology (ART). However, in the large amount of clinical practice we found remarkable variations in the effect of acupuncture among POI patients. The characteristics of POI patients who can benefit from acupuncture is unclear. In this study, POI patients will be treated by a validated acupuncture protocol in the actual medical environment via a registry study. The effect of acupuncture on the total patient population and each individual will be assessed using recognized outcome measurements. The high-throughout sequencing will be used to detect the serum differentially expressed miRNAs between POI patients benefited and not benefited from acupuncture. Bivariate correlation analysis will be used to explore the correlation between the differentially expressed miRNAs and the effect of acupuncture or the key factors influencing acupuncture effect. Those miRNAS showing a high correlation will be screened as predictive markers of acupuncture effect. Based on the above, the cutoff values of those candidate miRNAs in predicting acupuncture effect will be obtained via ROC curve. Results of this study will help the acupuncturists to distinguish POI patients who may benefit from acupuncture from those who may not, thus to improve their work efficiency and save the medical cost of POI patients insensitive to acupuncture.

早发性卵巢功能不全（POI）是导致不孕的重要原因，严重影响患者的生殖健康。目前现代医学尚无有效的方法恢复卵巢功能。研究表明，针刺可以改善POI患者的卵巢功能，增加其行辅助生殖技术助孕的机会。但基于大量的临床实践，我们发现POI患者的针刺疗效具有显著的个体差异，即针刺获益人群特征不明。本项目为病例注册登记研究，在实际医疗环境下，采用成熟的针刺方案干预POI患者，以公认的疗效指标评价针刺的总体疗效和患者的个体疗效。通过高通量测序分析针刺治疗有效和无效的POI患者血清miRNA的差异表达；通过二元变量相关分析探讨表达差异显著的miRNA与针刺疗效和/或其关键影响因素的相关性，筛选相关程度高的miRNA作为针刺疗效预测标志物；利用ROC曲线获得目标miRNA评价针刺疗效的有效界点。从而指导临床精准的选择POI针刺治疗的获益人群，提高针灸医师的工作效率，节省非针刺获益POI患者的医疗成本。

早发性卵巢功能不全（premature ovarian insufficiency, POI）是导致不孕的重要原因，严重影响患者的生殖健康。目前现代医学尚无有效的方法恢复卵巢功能。研究表明，针刺可以改善POI患者的卵巢功能，增加其行辅助生殖技术助孕的机会。基于大量的临床实践，我们发现POI患者的针刺疗效具有显著的个体差异，即针刺获益人群特征不明。本项目为病例注册登记研究，在实际医疗环境下，采用成熟的针刺方案干预POI患者，以公认的疗效指标评价针刺的总体疗效和患者的个体疗效。通过高通量测序分析针刺治疗有效和无效的POI患者血清miRNA的差异表达；通过二元变量相关分析探讨表达差异显著的miRNA与针刺疗效和/或其关键影响因素的相关性，筛选相关程度高的miRNA作为针刺疗效预测标志物；利用ROC曲线获得目标miRNA评价针刺疗效的有效界点。本课题纳入POI患者70.7%卵巢功能严重衰退，治疗难度大。研究结果提示，针刺可显著降低POI患者血清基础FSH水平，但其他卵巢功能评估指标（AFC、AMH、E2等）未见显著改善。POI患者单病例疗效评价显示，治疗12周有效病例占比45.7%，治疗24周有效病例占比28.57%。对13例患者（6例有效，3例无效，4例单时点有效）的39个血清样本进行了miRNA高通量测序，筛选出5个差异显著的microRNA：hsa-miR-3150a-3p、hsa-miR-3909、hsa-miR-497-5p、hsa-miR22-3p和miR229-3p，经Target基因关联分析及富集分析与细胞增殖的负调控有关。ROC曲线探索发现hsa-miR-497-5p可作为候选miRNA，其判断针刺治疗POI有效的截断值为0.03，敏感度为0.78，特异度为1。因样本量过少，以上结果尚需大样本验证。