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不明原因弱精子症患者精子中的 microRNA 谱分析。

MicroRNA profiling in spermatozoa of men with unexplained asthenozoospermia.

机构信息

Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

Legal Medicine Research Center, Legal Medicine Organization, Tehran, Iran.

出版信息

Andrologia. 2019 Jul;51(6):e13284. doi: 10.1111/and.13284. Epub 2019 Apr 22.

Abstract

Asthenozoospermia (AZS) which is characterised by decreased sperm motility is one of the main causes of male infertility. Recent studies demonstrated altered microRNAs (miRNAs) in total semen, seminal plasma and spermatozoa of asthenozoospermic men. In line with these studies, it was aimed to evaluate the miRNA expression profile in spermatozoa of unexplained asthenozoospermic men. Thirty-nine cases with idiopathic AZS and 35 fertile and healthy men as control were included. After total RNA extraction from spermatozoa, high-throughput sequencing technology was employed to display miRNA profiles in spermatozoa samples pooled from AZS cases and healthy controls. Relative quantification by real-time PCR was performed to validate RNA-seq results. SNORD48 was used as normaliser gene, and fold change was calculated by 2 method. Profiling results showed that 18 altered miRNAs in AZS men in comparison to controls. Subsequently, seven miRNAs were selected to validate by RT-PCR that showed MiR-888-3p significantly overexpressed in AZS cases (p = 0.014) in comparison with controls. It seems upregulation of miR-888-3p was associated with idiopathic AZS. This finding paves the way to the future investigation on the actual molecular role of miR-888-3p in aetiology of AZS.

摘要

弱精子症(AZS)的特征是精子运动能力下降,是男性不育的主要原因之一。最近的研究表明,在弱精子症男性的总精液、精浆和精子中存在改变的 microRNAs(miRNAs)。与这些研究一致,本研究旨在评估不明原因弱精子症男性精子中的 miRNA 表达谱。将 39 例特发性 AZS 病例和 35 例生育和健康男性作为对照组纳入研究。从精子中提取总 RNA 后,采用高通量测序技术显示精子样本的 miRNA 图谱,这些样本来自 AZS 病例和健康对照组。采用实时 PCR 进行相对定量以验证 RNA-seq 结果。SNORD48 作为正常化基因,通过 2 法计算倍数变化。分析结果显示,与对照组相比,AZS 男性有 18 个 miRNA 发生改变。随后,选择了 7 个 miRNA 通过 RT-PCR 进行验证,结果显示 MiR-888-3p 在 AZS 病例中明显过表达(p=0.014)。miR-888-3p 的上调似乎与特发性 AZS 有关。这一发现为进一步研究 miR-888-3p 在 AZS 病因学中的实际分子作用铺平了道路。

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