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隐睾固定术后非梗阻性无精子症患者睾丸中 miRNA 谱的改变。

Altered miRNA profile in testis of post-cryptorchidopexy patients with non-obstructive azoospermia.

机构信息

Reproductive Medicine Center, Department of Obstetrics and Gynecology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, People's Republic of China.

Anhui Province Key Laboratory of Reproductive Health and Genetics, Anhui Medical University, Hefei, Anhui, People's Republic of China.

出版信息

Reprod Biol Endocrinol. 2018 Aug 13;16(1):78. doi: 10.1186/s12958-018-0393-3.

Abstract

BACKGROUND

Cryptorchidism is one of the most common causes of non-obstructive azoospermia (NOA) leading to male infertility. Despite various medical approaches been utilised, many patients still suffer from infertility. MicroRNAs (miRNAs) play vital roles in the progress of spermatogenesis; however, little is known about the miRNA expression profile in the testes. Therefore, the miRNA profile was assessed in the testis of post-cryptorchidopexy patients.

METHODS

Three post-cryptorchidopexy testicular tissue samples from patients aged 23, 26 and 28 years old and three testis tissues from patients with obstructive azoospermia (controls) aged 24, 25 and 36 years old were used in this study. Next-generation sequencing (NGS) was used to perform the miRNA expression profiling. Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) assays were subsequently used to confirm the results of several randomly-selected and annotated miRNAs.

RESULTS

A series of miRNAs were found to be altered between post-cryptorchidopexy testicular tissues and control tissues, including 297 downregulated and 152 upregulated miRNAs. In the subsequent qRT-PCR assays, the expression levels of most of the selected miRNAs (9/12, P < 0.05) were consistent with the results of NGS technology. Furthermore, signal transduction, adaptive immune response and biological regulation were associated with the putative target genes of the differentially-expressed miRNAs via GO analysis. In addition, oxidative phosphorylation, Parkinson's disease and ribosomal pathways were shown to be enriched using KEGG pathway analysis of the differentially-expressed genes.

CONCLUSIONS

This study provides a global view of the miRNAs involved in post-cryptorchidopexy testicular tissues as well as the altered expression of miRNAs compared to control tissues, thus confirming the vital role of miRNAs in cryptorchidism.

摘要

背景

隐睾是导致男性不育的无梗阻性无精子症(NOA)的最常见原因之一。尽管已经采用了各种医学方法,但许多患者仍然患有不育症。微小 RNA(miRNA)在精子发生过程中起着至关重要的作用;然而,miRNA 在睾丸中的表达谱知之甚少。因此,评估了隐睾松解术后患者睾丸中的 miRNA 表达谱。

方法

本研究使用了 3 名年龄分别为 23、26 和 28 岁的隐睾松解术后患者的睾丸组织样本和 3 名年龄分别为 24、25 和 36 岁的梗阻性无精子症患者(对照组)的睾丸组织样本。采用下一代测序(NGS)进行 miRNA 表达谱分析。随后使用定量实时逆转录聚合酶链反应(qRT-PCR)检测来验证随机选择和注释的几个 miRNA 的结果。

结果

在隐睾松解术后睾丸组织和对照组组织之间发现了一系列 miRNA 发生改变,包括 297 个下调和 152 个上调 miRNA。在随后的 qRT-PCR 检测中,大多数选定 miRNA 的表达水平(9/12,P<0.05)与 NGS 技术的结果一致。此外,GO 分析显示,通过差异表达 miRNA 的假定靶基因,信号转导、适应性免疫反应和生物调节与这些基因相关。此外,KEGG 通路分析显示,差异表达基因中存在氧化磷酸化、帕金森病和核糖体途径的富集。

结论

本研究提供了隐睾松解术后睾丸组织中涉及的 miRNA 的全面视图,以及与对照组组织相比 miRNA 表达的改变,从而证实了 miRNA 在隐睾症中的重要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac54/6090726/efded309ccce/12958_2018_393_Fig1_HTML.jpg

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