Effects of miR-98 in intrauterine extracellular vesicles on maternal immune regulation during the peri-implantation period in cattle
Effects of miR-98 in intrauterine extracellular vesicles on maternal immune regulation during the peri-implantation period in cattle
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DOI：
10.1038/s41598-019-56879-w
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发表时间：
2019-12-30
影响因子：
4.6
通讯作者：
Imakawa, Kazuhiko
中科院分区：
文献类型：
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作者：
Nakamura, Keigo;Kusama, Kazuya;Imakawa, Kazuhiko
越来越多的证据表明，细胞外囊泡（EVs）存在于子宫腔中发挥作用的概念，子宫内膜细胞的相互作用在围着床期。然而，子宫内EV如何对子宫内膜起作用尚未得到很好的表征。为了研究子宫内EV如何影响牛的子宫内膜环境，在妊娠第17或20天（分别为P17、P20;孕体植入子宫内膜开始于第19-19.5天）用从子宫冲洗液（UF）中分离的EV处理牛子宫内膜上皮细胞（EEC）。然后对从EECs提取的RNA进行RNA序列分析。结果表明，与P17相比，P20时EV处理的EECs中与免疫系统相关的转录物表达下调。为了研究EV中的microRNA（miRNAs）是否在围着床期调节子宫内膜中的母体免疫系统，进行microRNA序列和计算机分析，鉴定EV中的bta-miR-98作为调节母体免疫系统的潜在miRNA。此外，用bta-miR-98处理EECs负调节了EECs中的几种免疫系统相关基因，CTSC，IL 6，CASP 4和IKBKE。这些结果表明，含有bta-miR-98的EV是母体免疫系统的调节剂，可能允许孕体在围着床期附着于子宫内膜上皮。
Evidence accumulated suggests that extracellular vesicles (EVs) present in uterine lumen play a role in conceptus-endometrial cell interactions during peri-implantation periods. However, how intrauterine EVs function on endometrium have not been well characterized. To study how intrauterine EVs affect endometrial milieu in cattle, bovine endometrial epithelial cells (EECs) were treated with EVs isolated from uterine flushing fluids (UFs) on day 17 or 20 pregnancy (P17, P20, respectively; conceptus implantation to endometrium begins on days 19-19.5). RNA extracted from EECs were then subjected to RNA sequence analysis. The analysis revealed that transcripts related to immune system were down-regulated in EECs treated with EVs on P20 compared with those on P17. To investigate whether microRNAs (miRNAs) in EVs regulate maternal immune system in the endometrium during the peri-implantation, microRNA sequence and in silico analyses were performed, identifying bta-miR-98 in EVs as a potential miRNA to regulate maternal immune system. Furthermore, the treatment of EECs with bta-miR-98 negatively regulated several immune system-related genes, CTSC, IL6, CASP4 and IKBKE, in EECs. These results suggest that EVs containing bta-miR-98 is a regulator of maternal immune system, possibly allowing the conceptus attachment to the endometrial epithelium during the peri-implantation period.