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死后血清中外泌体的稳定性及心肌梗死尸检血清中外泌体 miRNA 表达谱的初步研究。

Stability of exosomes in the postmortem serum and preliminary study on exosomal miRNA expression profiling in serum from myocardial infarction cadavers.

机构信息

Department of Forensic Medicine, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-Cho, Mizuho-Ku, Nagoya, 467-8601, Japan.

出版信息

Int J Legal Med. 2023 May;137(3):825-834. doi: 10.1007/s00414-022-02913-y. Epub 2022 Nov 23.

Abstract

Exosome-encapsulated miRNAs could potentially be sensitive biomarkers of human diseases. Since a lipid bilayer membrane surrounds exosomes, the exosomal miRNA may stably exist in body fluids with diseases as well as biological fluids. Therefore, exosomal miRNA may be helpful for autopsy diagnosis. Assuming cadaver blood would be most useful, we initially examined serum exosome stability with regard to storage temperatures and periods. Characteristic analyses of the exosome revealed that exosomes and the content, miRNA, were stably preserved until at least three days when stored at below 20 °C. Subsequently, exosomal miRNA expression profiling was performed on the serum of acute myocardial infarction (AMI, 4 cases) autopsy bodies and on hemorrhagic shock bodies used as the control (CT, 3 cases). Results showed that significant twofold up- and downregulations of expression of 18 and 16 miRNAs were detectable in AMI as compared to the CT, respectively. miR-126-3p, which has been reported to be increased in serum of AMI patients and a mouse model, was one of the significantly upregulated miRNAs. Furthermore, dysregulation of exosomal miRNAs, such as miR-145-5p, miR-143-3p, and miR-222-3p, which are involved in cardioprotection, may be associated with AMI pathogenesis. These findings provide a novel perspective on the potential role of exosomal miRNA in determining the cause of death.

摘要

外泌体包裹的 miRNA 可能成为人类疾病的敏感生物标志物。由于脂质双层膜包围着外泌体,因此外泌体中的 miRNA 可以在疾病相关的体液以及生物体液中稳定存在。因此,外泌体 miRNA 可能有助于尸检诊断。假设尸检血液最有用,我们最初检查了储存温度和时间对血清外泌体稳定性的影响。对外泌体的特征分析表明,外泌体及其内容物 miRNA 至少在 3 天内稳定保存,储存温度低于 20°C。随后,对急性心肌梗死 (AMI) 尸检患者血清和作为对照的失血性休克患者 (CT) 的血清进行了外泌体 miRNA 表达谱分析 (n=4 和 n=3)。结果显示,与 CT 相比,AMI 中 18 个 miRNA 的表达显著上调了两倍,16 个 miRNA 的表达显著下调了两倍。miR-126-3p 是一种已知在 AMI 患者和小鼠模型血清中升高的 miRNA,是上调最为显著的 miRNA 之一。此外,外泌体 miRNA 的失调,如 miR-145-5p、miR-143-3p 和 miR-222-3p,这些 miRNA 参与了心脏保护,可能与 AMI 发病机制有关。这些发现为外泌体 miRNA 在确定死因方面的潜在作用提供了新的视角。

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