Potential confounding factors in measurement of specific cell-free seminal mRNAs and microRNAs derived from human reproductive organs
Potential confounding factors in measurement of specific cell-free seminal mRNAs and microRNAs derived from human reproductive organs
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测量源自人类生殖器官的特定无细胞精液 mRNA 和 microRNA 时的潜在混杂因素
DOI：
10.1111/andr.12238
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发表时间：
2016-11-01
期刊：
影响因子：
4.5
通讯作者：
Li, H. G.
中科院分区：
文献类型：
--
作者：
Dong, T. T.;Yu, Q.;Li, H. G.
无细胞精液RNA(CFS-RNA)是一种来源于男性生殖器官的混合转录本，是研究和诊断男性生殖相关疾病的潜在生物标志物。然而，一些临床因素，包括年龄、无症状解脲支原体(UU)感染、阴囊热应激、禁欲时间和精液标本的保存条件，可能会干扰精子参数和精液生物标志物的测定。因此，本研究旨在评价上述临床因素对CFS-RNA测定的影响，为CFS-RNA的研究应用和潜在的临床应用奠定基础。根据选定的临床因素采集精液样本。通过离心法获得无细胞精浆，用TRIzol LS提取总RNA。用实时定量聚合酶链式反应对选择性雄性生殖器官特异性CFS-mRNAs和CFS-miRNAs进行定量。计算并比较一次射精液中CFS-mRNAs和CFS-miRNAs的浓度和总量。阴囊热应激后ACTB、DDX4、WFDC9和miR-514a-3p均显著升高。SEMG1(精囊特异性)随戒断时间延长呈下降趋势。年龄、无症状解脲支原体感染和储存条件对CFS-RNA的测定无显著影响。这些结果表明，阴囊热应激显著干扰精囊和睾丸来源的CFS-RNA的选择，禁欲时间可能影响精囊CFS-RNA的产量，而其他临床因素对其影响不显著。因此，在CFS-RNA的研究或临床应用中，应考虑热暴露和禁欲时间。
Cell-free seminal RNA (cfs-RNA) is mixed transcripts derived from male reproductive organs, and is potential biomarker for the research and diagnosis of male reproductive-related diseases. However, some clinical factors, including age, asymptomatic Ureaplasma urealyticum (UU) infection, scrotal heat stress, abstinence period, and the storage condition of semen samples, may interfere with sperm parameters and the measurement of seminal biomarkers. Accordingly, this study was designed to evaluate the effect of above clinical factors on the measurement of cfs-RNA, aiming to lay a foundation for its research use and potential clinical application. Semen samples were collected according to the selected clinical factors. Cell-free seminal plasma was obtained by centrifugation and total RNA was extracted with TRIzol LS. Selective male reproductive organ-specific cfs-mRNAs and cfs-miRNAs were quantified by quantitative real-time PCR. The concentration and total amount of cfs-mRNAs and cfs-miRNAs in one ejaculate were calculated and compared. ACTB, DDX4 (testis-specific), WFDC9 (epididymis-specific), and miR-514a-3p (testis-specific) significantly increased after scrotal heat stress. SEMG1 (seminal vesicle-specific) showed declining tendency with the prolonged abstinence period. Age, asymptomatic UU infection, and the storage condition showed no significant impact on the measurement of cfs-RNA. These results indicate that scrotal heat stress significantly interfere with the selected cfs-RNA derived from the testis and epididymis, and abstinence period may affect the yield of cfs-mRNA from seminal vesicle, while other clinical factors has no significant impact on the measurement. Thus, heat exposure and abstinence period should be considered for the cfs-RNA measurement in its research or clinical application.