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脂多糖诱导流产小鼠模型中胎盘来源外泌体中miR-17和miR-29a的表达:一项实验研究。

The expression of miR-17 and miR-29a in placenta-derived exosomes in LPS-induced abortion mice model: An experimental study.

作者信息

Jalilvand Tahereh, Salarinia Reza, Ahmadabad Hasan Namdar, Safdari Mohammadreza

机构信息

Student Research Committee, School of Medicine, North Khorasan University of Medical Sciences, Bojnurd, Iran.

Department of Advanced Sciences and Technologies, School of Medicine, North Khorasan University of Sciences, Bojnurd, Iran.

出版信息

Int J Reprod Biomed. 2021 Jun 23;19(5):433-440. doi: 10.18502/ijrm.v19i5.9252. eCollection 2021 May.

Abstract

BACKGROUND

The expression pattern of microRNAs in placenta-derived exosomes plays a crucial role in the regulation of immune responses and inflammation at the fetal-maternal interface.

OBJECTIVE

Considering the immunomodulatory properties of miR-17 and miR-29a, we determined their expression levels in placenta-derived exosomes in a lipopolysaccharide (LPS)-induced abortion mice model.

MATERIALS AND METHODS

A total of 14 pregnant BALB/c mice, aged 6-8 wk, were randomly divided into two groups (n = 7/each) on the gestational day 11.5. While the mice in the experimental group were treated with LPS, those in the control group were treated with Phosphate buffered saline; 5 hr after the treatment, the placental cells were isolated and cultured for 48 hr. Then, the cell culture supernatants were collected and used for isolation of exosomes. The isolated exosomes were confirmed by western blot and scanning electron microscopy. The miRNAs were then extracted from exosomes, and cDNA synthesized. The expression levels of miR-17 and miR-29a were evaluated by quantitative real-time PCR analysis.

RESULTS

Our results showed that the expression levels of miR-29a in placenta-derived exosomes obtained from the experimental group increased significantly compared to the control group. Also, the expression levels of miR-17 in the placenta-derived exosomes obtained from the experimental group were found to decrease; however, it did not show significant changes compared with the control group (p > 0.05).

CONCLUSION

Inflammatory reactions at the fetal-maternal interface can alter miRNAs expression patterns in placenta-derived exosomes, especially miRNAs with immunomodulatory effects such as miR-29a.

摘要

背景

微小RNA在胎盘来源外泌体中的表达模式在母胎界面免疫反应和炎症调节中起关键作用。

目的

鉴于miR-17和miR-29a的免疫调节特性,我们在脂多糖(LPS)诱导流产的小鼠模型中测定了它们在胎盘来源外泌体中的表达水平。

材料与方法

选取14只6 - 8周龄的妊娠BALB/c小鼠,于妊娠第11.5天随机分为两组(每组n = 7)。实验组小鼠用LPS处理,对照组小鼠用磷酸盐缓冲盐水处理;处理5小时后,分离胎盘细胞并培养48小时。然后收集细胞培养上清液用于外泌体分离。通过蛋白质免疫印迹法和扫描电子显微镜对分离得到的外泌体进行鉴定。随后从外泌体中提取微小RNA并合成cDNA。通过定量实时PCR分析评估miR-17和miR-29a的表达水平。

结果

我们的结果显示,与对照组相比,实验组胎盘来源外泌体中miR-29a的表达水平显著升高。此外,发现实验组胎盘来源外泌体中miR-17的表达水平降低;然而,与对照组相比未显示出显著变化(p > 0.05)。

结论

母胎界面的炎症反应可改变胎盘来源外泌体中的微小RNA表达模式,尤其是具有免疫调节作用的微小RNA,如miR-29a。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d982/8261093/dc8ab295fccd/ijrb-19-433-g001.jpg

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