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人源支持细胞来源的细胞外囊泡:鉴定、蛋白质组学分析和 miRNA 谱分析。

Extracellular vesicles derived from human Sertoli cells: characterizations, proteomic analysis, and miRNA profiling.

机构信息

Department of Urology, Peking University First Hospital, Beijing, 100034, People's Republic of China.

Institute of Urology, Peking University, Beijing, 100034, People's Republic of China.

出版信息

Mol Biol Rep. 2022 Jun;49(6):4673-4681. doi: 10.1007/s11033-022-07316-1. Epub 2022 Apr 2.

Abstract

BACKGROUND

Extracellular vesicles (EVs) contain thousands of proteins and nucleic acids, playing an important role in cell-cell communications. Sertoli cells have been essential in the testis as a "nurse cell". However, EVs derived from human Sertoli cells (HSerCs) have not been well investigated.

METHODS

EVs were isolated from HSerCs via ultracentrifugation and characterized by transmission electron microscopy, tunable resistive pulse sensing, and Western blotting. The cargo carried by HSerCs-EVs was measured via liquid chromatography-mass spectrometry and GeneChip miRNA Arrays. Bioinformatic analysis was performed to reveal potential functions of HSerCs-EVs.

RESULTS

A total of 860 proteins with no less than 2 unique peptides and 88 microRNAs with high signal values were identified in HSerCs-EVs. Biological processes related to molecular binding, enzyme activity, and regulation of cell cycle were significantly enriched. Specifically, many proteins in HSerCs-EVs were associated with spermatogenesis and regulation of immune system, including Septins, Large proline-rich protein BAG6, Clusterin, and Galectin-1. Moreover, abundant microRNAs within HSerCs-EVs (miR-638, miR-149-3p, miR-1246, etc.) had a possible impact on male reproductive disorders such as asthenozoospermia and oligozoospermia.

CONCLUSIONS

Our study has shown that HSerCs-EVs contain diverse components such as proteins and microRNAs. Further research is required to evaluate HSerCs-EVs in spermatogenesis, which are underutilized but highly potent resources with particular promise for male infertility.

摘要

背景

细胞外囊泡(EVs)包含数千种蛋白质和核酸,在细胞间通讯中发挥着重要作用。Sertoli 细胞作为“滋养细胞”在睾丸中起着至关重要的作用。然而,人 Sertoli 细胞来源的 EVs(HSerCs-EVs)尚未得到充分研究。

方法

通过超速离心从 HSerCs 中分离 EVs,并通过透射电子显微镜、可调电阻脉冲感应和 Western blot 进行表征。通过液相色谱-质谱和 GeneChip miRNA 阵列测量 HSerCs-EVs 携带的货物。进行生物信息学分析以揭示 HSerCs-EVs 的潜在功能。

结果

在 HSerCs-EVs 中鉴定出总共 860 种具有不少于 2 个独特肽的蛋白质和 88 种具有高信号值的 microRNAs。与分子结合、酶活性和细胞周期调节相关的生物学过程显著富集。具体而言,HSerCs-EVs 中的许多蛋白质与精子发生和免疫系统调节有关,包括 Septins、Large proline-rich protein BAG6、Clusterin 和 Galectin-1。此外,HSerCs-EVs 中丰富的 microRNAs(miR-638、miR-149-3p、miR-1246 等)可能对男性生殖障碍如弱精症和少精症产生影响。

结论

我们的研究表明,HSerCs-EVs 包含多种成分,如蛋白质和 microRNAs。需要进一步研究以评估 HSerCs-EVs 在精子发生中的作用,这是一种未充分利用但具有巨大潜力的资源,对男性不育症具有特殊的应用前景。

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