男性不育症患者精浆piR-32678的表达及其临床意义

Expression of piR-32678 in seminal plasma of the patients with male infertility and its clinical significance

目的获得非梗阻性无精子症(NOA)患者及精液参数正常男性精浆piRNA表达谱,探讨差异表达的piR-32678作为男性不育分子诊断标记物的可行性。方法对健康对照者与NOA患者各5例的精浆标本进行piRNA芯片检测。通过实时荧光定量PCR(RT-qPCR)对26例健康对照者、18例少精子症患者、7例NOA患者及11例梗阻性无精子症(OA)患者精浆标本验证piR-32678水平。对精浆piR-32678在少精子症中的诊断价值进行ROC曲线分析。结果芯片结果显示,3652种piRNA在NOA患者精浆中的表达量是健康对照者变化的2倍以上。qRT-PCR验证结果显示,与健康对照者相比,少精子症组和NOA组精浆中piR-32678的相对表达量显著上调;而OA组精浆中piR-32678的相对表达量差异无统计学意义(P>0.05)。受试者工作特征曲线(ROC曲线)分析piR-32678诊断少精子症的曲线下面积为0.950(95%CI,0.890~1.000)。结论少精子症患者和NOA患者精浆piR-32678表达水平与精液健康男性间存在显著差异。精浆piR-32678有望成为男性不育评估的无创分子标记物,并为男性不育症的病因研究提供新的思路。

Objective To obtain the seminal plasma piRNA profiles in patients with non-obstructive azoospermia(NOA)and males with healthy semen analysis parameters,and investigate the potential of the differentially expressed piR-32678 as a novel molecular biomarker for the diagnosis of male infertility.Methods The piRNA microarray was performed on seminal plasma samples from 5 patients with NOA and 5 males with normal semen analysis parameters.The relative expression of seminal plasma piR-32678 was verified by RT-qPCR on 26 normal controls,18 oligospermia,7 NOA and 11 obstructive azoospermia(OA)patients.The diagnostic value of seminal plasma piR-32678 in the oligospermia was evaluated by ROC curve.Results Microarray results showed that there were 3652 kinds of piRNAs in seminal plasma exhibited more than 2-fold change between the patients with NOA and the controls.The verification result of qRT-PCR showed that compared with the healthy control group,the relative expression of seminal plasma piR-32678 in oligospermia group and in the NOA group was significantly up-regulated.However,there was no significant difference in that of OA group(P>0.05).ROC curve analysis showed that AUC of piR-32678 for diagnosing oligospermia was 0.950(95%CI,0.890-1.000).Conclusion The expression of piR-32678 in seminal plasma of the patients with oligospermia and with NOA is significantly different from that of men with healthy semen.Seminal plasma piR-32678 is expected to be a noninvasive molecular maker for the evaluation of male infertility,and might provides a new idea for the study on the etiology of male infertility.