羊水中的外泌体促进人精子获能。

Exosomes from uterine fluid promote capacitation of human sperm.

机构信息

Department of Urology, The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, China.

Department of Genetics, The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, China.

出版信息

PeerJ. 2024 Apr 25;12:e16875. doi: 10.7717/peerj.16875. eCollection 2024.

DOI:10.7717/peerj.16875

PMID:38680889

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11056104/

Abstract

BACKGROUND

Extracellular vesicles (EVs) are membrane-bound vesicles containing various proteins, lipids, and nucleic acids. EVs are found in many body fluids, such as blood and urine. The release of EVs can facilitate intercellular communication through fusion with the plasma membrane or endocytosis into the recipient cell or through internalization of the contents. Recent studies have reported that EVs isolated from human endometrial epithelial cells (EECs) promote sperm fertilization ability. EVs from uterine flushing fluid more closely resemble the physiological condition of the uterus. However, it is unclear whether EVs derived directly from uterine flushing fluid have the same effect on sperm. This study aimed to research the effect of EVs from uterine flushing fluid on sperm.

METHODS

EVs were isolated from the uterine flushing fluid. The presence of EVs was confirmed by nanoparticle tracking analysis (NTA), Western blot, and transmission electron microscopy (TEM). EVs were incubated with human sperm for 2 h and 4 h. The effects of EVs on sperm were evaluated by analyzing acrosome reaction, sperm motility, and reactive oxygen species (ROS).

RESULTS

The EVs fractions isolated from the uterine fluid were observed in cup-shaped vesicles of different sizes by TEM. All isolated vesicles contained similar numbers of vesicles in the expected size range (30-200 nm) by NTA. CD9 and CD63 were detected in EVs by western blot. Comparing the motility of the two groups incubated sperm motility significantly differed at 4 h. The acrosome reactions were promoted by incubating with EVs significantly. ROS were increased in sperm incubated with EVs.

CONCLUSION

Our results showed EVs present in the uterine fluid. Acrosome reactions and ROS levels increased in human sperm incubated with EVs. EVs from uterine fluid can promote the capacitation of human sperm. The increased capacitation after sperm interaction with EVs suggests a possible physiological effect during the transit of the uterus.

摘要

背景

细胞外囊泡（EVs）是含有各种蛋白质、脂质和核酸的膜结合囊泡。EVs 存在于许多体液中，如血液和尿液。EVs 通过与质膜融合或内吞作用进入受体细胞或通过内容物内化来促进细胞间通讯。最近的研究报告称，从人子宫内膜上皮细胞（EECs）中分离出的 EVs 可提高精子受精能力。来自子宫冲洗液的 EVs 更接近子宫的生理状态。然而，尚不清楚直接来自子宫冲洗液的 EVs 是否对精子有相同的作用。本研究旨在研究子宫冲洗液中 EVs 对精子的影响。

方法

从子宫冲洗液中分离 EVs。通过纳米颗粒跟踪分析（NTA）、Western blot 和透射电子显微镜（TEM）证实 EVs 的存在。将 EVs 与人精子孵育 2 小时和 4 小时。通过分析顶体反应、精子运动和活性氧（ROS）来评估 EVs 对精子的影响。

结果

通过 TEM 观察到从子宫液中分离出的 EVs 呈不同大小的杯状囊泡。所有分离的囊泡均通过 NTA 检测到预期大小范围内（30-200nm）的囊泡数量相似。Western blot 检测到 EVs 中的 CD9 和 CD63。与对照组相比，孵育 4 小时后两组精子的运动能力有显著差异。与 EVs 孵育可显著促进顶体反应。与 EVs 孵育的精子中 ROS 增加。