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从高度纯化的高密度脂蛋白中高通量分离微小RNA的快速简易方法

Fast and Simplified Method for High Through-put Isolation of miRNA from Highly Purified High Density Lipoprotein.

作者信息

Seneshaw Mulugeta, Mirshahi Faridoddin, Min Hae-Ki, Asgharpour Amon, Mirshahi Shervin, Daita Kalyani, Boyett Sherry, Santhekadur Prasanna K, Fuchs Michael, Sanyal Arun J

机构信息

Division of Gastroenterology, Hepatology and Nutrition, Virginia Commonwealth University.

McGuire Veterans Affairs Medical Center, Research BC-101.

出版信息

J Vis Exp. 2016 Jul 27(113):54257. doi: 10.3791/54257.

Abstract

Small non-coding RNAs (miRNAs) have been implicated in a variety of human diseases including metabolic syndromes. They may be utilized as biomarkers for diagnosis and prognosis or may serve as targets for drug development, respectively. Recently it has been shown that miRNAs are carried in lipoproteins, particularly high density lipoproteins (HDL) and are delivered to recipient cells for uptake. This raises the possibility that miRNAs play a critical and pivotal role in cellular and organ function via regulation of gene expression as well as messenger for cell-cell communications and crosstalk between organs. Current methods for miRNA isolation from purified HDL are impractical when utilizing small samples on a large scale. This is largely due to the time consuming and laborious methods used for lipoprotein isolation. We have developed a simplified approach to rapidly isolate purified HDL suitable for miRNA analysis from plasma samples. This method should facilitate investigations into the role of miRNAs in health and disease and in particular provide new insights into the variety of biological functions, outside of the reverse cholesterol transport, that have been ascribed to HDL. Also, the miRNA species which are present in HDL can provide valuable information of clinical biomarkers for diagnosis of various diseases.

摘要

小非编码RNA(miRNA)与包括代谢综合征在内的多种人类疾病有关。它们可分别用作诊断和预后的生物标志物,或作为药物开发的靶点。最近有研究表明,miRNA存在于脂蛋白中,尤其是高密度脂蛋白(HDL),并被递送至受体细胞进行摄取。这增加了一种可能性,即miRNA通过调节基因表达以及作为细胞间通讯和器官间相互作用的信使,在细胞和器官功能中发挥关键和核心作用。当大规模使用小样本时,目前从纯化的HDL中分离miRNA的方法并不实用。这主要是由于用于脂蛋白分离的方法耗时且费力。我们开发了一种简化方法,可快速从血浆样本中分离出适合miRNA分析的纯化HDL。该方法应有助于研究miRNA在健康和疾病中的作用,特别是为HDL除逆向胆固醇转运之外的多种生物学功能提供新的见解。此外,HDL中存在的miRNA种类可为各种疾病的诊断提供有价值的临床生物标志物信息。

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本文引用的文献

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Characterization of levels and cellular transfer of circulating lipoprotein-bound microRNAs.循环脂蛋白结合 microRNAs 水平和细胞转移的特征。
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Extracellular microRNA: a new source of biomarkers.细胞外 microRNA:一种新的生物标志物来源。
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