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当前我国有15%左右的育龄夫妇面临不孕不育症的问题,而其中约一半因素由男性不育引起[1-2]。原发性无精子症(idiopathic azoospermia,IA)是指男性精液中表现出不明原因的无精子,而细胞遗传学和激素内分泌检查均正常[3-4]。IA是男性不育的最严重形式之一,发病率占男性不育的30%~40%[5]。尽管研究者在男性不育家族病例及动物模型中发现一些基因异常表达与IA密切相关,但IA的潜在机制仍不清楚。微小RNA(miRNA)是一类小的非编码RNA分子,通常由19~23个核苷酸构成,通过与目的基因信使RNA(mRNA)互补结合来降解或抑制目的基因翻译[6]。miRNA在生理和病理过程中具有广泛的功能,并已阐明其在调节精子发生方面亦发挥着重要作用[7]。一些miRNA可参与减数分裂中关键基因的沉默,当这些miRNA表达发生改变时则导致男性不育[8]。ZHUANG等[9]通过芯片发现51个miRNA在IA病人睾丸组织中表达上调,而42个miRNA表达下调,其中miR-186表达上调超过4倍。目前,关于miR-186与IA的相关性研究尚未见报道。本研究收集IA病人140例和正常生育的男性140名的外周血标本,旨在检测miR-186在2组外周中的表达水平,分析其与无精子症因子(AZF)微缺失的相关性,并评估其在IA诊断中的价值。
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2组年龄、体质量指数、睾酮水平及吸烟情况差异均无统计学意义(P>0.05)。IA组中8例病人存在AZF微缺失,微缺失率为5.71 %(8/140),而对照组未检测出AZF微缺失(P <0.05)(见表 1)。
分组 n 年龄/岁 体质量指数/
(kg/m2)睾酮水平/
(ng/dL)吸烟
情况AZF
微缺失对照组 140 30.42±4.89 23.66±2.30 364.92±63.78 48 0 IA组 140 31.06±5.17 23.48±2.65 377.16±55.41 52 8 t — 0.95 0.42 1.71 0.25△ 6.31* P — > 0.05 > 0.05 > 0.05 > 0.05 < 0.05 △示χ2值;*示校正χ2值 表 1 2组一般资料比较(x±s)
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miR-186在对照组外周血中的表达量为0.28±0.21,而在IA组外周血中的表达量为0.75±0.42,2组间miR-186表达量差异有统计学意义(t=11.84,P <0.01)。
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根据IA组中病人AZF微缺失情况,将其分为AZF微缺失组(n=8)和非AZF微缺失组(n=132)。miR-186在AZF微缺失组中的表达量为0.76±0.24,而在非AZF微缺失组中的表达量为0.72±0.40,2组间miR-186表达差异无统计学意义(t=0.28,P>0.05)。Spearman相关分析显示,外周血中miR-186的表达与AZF微缺失无相关性(P>0.05)。
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采用ROC曲线评价外周血中miR-186表达对IA的诊断价值。miR-186的ROC曲线下面积为0.84(95% CI: 0.794~0.886)。当miR-186表达量为0.52时为最佳诊断分界点,敏感度为71.86 %,特异度为80.39 %(见图 1)。
微小RNA-186在原发性无精子症病人外周血中的表达及诊断价值
Expression and diagnostic value of microRNA-186 in peripheral blood of patients with idiopathic azoospermia
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摘要:
目的检测微小RNA-186(miR-186)在原发性无精子症(IA)病人外周血中的表达量,并探讨其对IA的诊断价值。 方法收集2015-2017年在泌尿外科就诊的IA病人(IA组)140例和正常生育的男性(对照组)140名的外周血标本。常规提取外周血DNA和RNA,采用多重PCR试验检测无精子症因子(AZF)在2组间的微缺失情况。采用实时荧光定量PCR试验检测miR-186在2组间的表达差异。分析外周血中miR-186的表达与AZF微缺失的相关性。采用ROC分析评价外周血中miR-186表达对IA的诊断价值。 结果2组年龄、体质量指数、睾酮水平及吸烟情况差异均无统计学意义(P>0.05)。IA组中8例病人存在AZF微缺失(P < 0.05),微缺失率为5.71%(8/140),而对照组中未检测出AZF微缺失(P < 0.05)。miR-186在IA组外周血中的表达量较对照组明显升高(P < 0.01)。外周血中miR-186的表达与AZF微缺失无相关性(P>0.05)。miR-186对IA具有较高的诊断价值,ROC曲线下面积为0.84(95% CI:0.794~0.886),当miR-186表达量为0.52时为最佳诊断分界点,敏感度为71.86%,特异度为80.39%。 结论miR-186在IA病人外周血中高表达,但与AZF微缺失无相关性。外周血中高表达的miR-186可能作为IA的一种新的诊断标志物。 Abstract:ObjectiveTo detect the expression levels of microRNA-186(miR-186) in the peripheral blood of patients with idiopathic azoospermia(IA), and investiagte its diagnostic value for IA. MethodsThe peripheral blood samples from 140 IA patients from 2015 to 2017 in urology department and 140 normal fertile men were divided into the IA group and control group, respectively.The DNA and RNA in two groups were routinely extracted from peripheral blood, and the microdeletion of azoospermia factor(AZF) in two groups were detected using multiplex PCR.The expression levels of miR-186 in two groups were detected using quantitative real-time PCR.The correlation between miR-186 expression and AZF microdeletion in peripheral blood was analyzed.The diagnostic value of miR-186 for IA in peripheral blood was evaluated by ROC analysis. ResultsThe differences of the age, body mass index, testosterone level and smoking status were not statistically significant between two groups(P>0.05).Eight patients with AZF microdeletion in the IA group were found, and the microdeletion rate of which was 5.71%(8/140), while no AZF microdeletion was detected in the control group(P>0.05).The expression level of miR-186 in IA group was significantly higher than that in control group(P < 0.01).The miR-186 expression in peripheral blood was not correlated with AZF microdeletion(P>0.05).The miR-186 had a higher diagnosis value for IA, and the area under the ROC curve was 0.84(95% CI: 0.79 to 0.87).The expression level of miR-186 at 0.52 was the best diagnostic cut-off point, and the sensitivity and specificity were 71.86% and 80.39%, respectively. ConclusionsThe expression level of miR-186 is high in the peripheral blood of IA patients, but its expression is not correlated with AZF microdeletion.High expression of miR-186 in peripheral blood may serve as a new diagnostic biomarker for IA. -
Key words:
- idiopathic azoospermia /
- microRNA-186 /
- microdeletion /
- peripheral blood /
- biomarker
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表 1 2组一般资料比较(x±s)
分组 n 年龄/岁 体质量指数/
(kg/m2)睾酮水平/
(ng/dL)吸烟
情况AZF
微缺失对照组 140 30.42±4.89 23.66±2.30 364.92±63.78 48 0 IA组 140 31.06±5.17 23.48±2.65 377.16±55.41 52 8 t — 0.95 0.42 1.71 0.25△ 6.31* P — > 0.05 > 0.05 > 0.05 > 0.05 < 0.05 △示χ2值;*示校正χ2值 -
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