OIP5-AS1通过靶向miR-140-5p调控RhoA/ROCK2信号通路，促进Th17细胞分化和实验性自身免疫性脑脊髓炎的严重程度。

OIP5-AS1 facilitates Th17 differentiation and EAE severity by targeting miR-140-5p to regulate RhoA/ROCK2 signaling pathway.

作者信息

Liu Ruihua, Li Yan, Zhou Haitao, Wang Hao, Liu Dequan, Wang Huilin, Wang Zhenghua

机构信息

Department of Neurology, Luoyang Central Hospital Affiliated to Zhengzhou University, Luoyang, China.

出版信息

Life Sci. 2021 Jan 27:119108. doi: 10.1016/j.lfs.2021.119108.

DOI:10.1016/j.lfs.2021.119108

PMID:33515560

Abstract

AIMS

Multiple sclerosis (MS) is one of the commonest neurologic disorders globally. LncRNA OIP5-AS1 has been found to be implicated in the etiology of MS. This study was to explore the roles and molecular mechanisms of OIP5-AS1 in the development of MS.

MATERIALS AND METHODS

RT-qPCR assay was used to measure expressions of OIP5-AS1, miR-140-5p, IL-17A mRNA and RhoA mRNA. CD4IL-17 cell proportion was determined by flow cytometry. IL-17A secretion was examined by ELISA assay. Cell inflammatory infiltration and demyelination were assessed by histological analyses. The interaction between miR-140-5p and OIP5-AS1 or RhoA 3'UTR was validated by bioinformatical analysis and luciferase reporter assay. Western blot assay was performed to detect protein expressions of ROCK2 and RhoA. An experimental autoimmune encephalomyelitis (EAE) models was established to explore the role of OIP5-AS1 in MS in vivo.

KEY FINDINGS

OIP5-AS1 expression was enhanced in MS patients. Also, elevated OIP5-AS1 level was observed during T-helper 17 (Th17) differentiation. Moreover, OIP5-AS1 promoted Th17 differentiation in vitro and contributed to the development of EAE in vivo. Mechanical explorations revealed that OIP5-AS1 targeted miR-140-5p to regulate Th17 differentiation. Moreover, RhoA was a target of miR-140-5p and miR-140-5p inhibited the activation of RhoA/ROCK2 signaling. Also, RhoA upregulation abrogated the inhibitory effects of miR-140-5p on Th17 differentiation.

SIGNIFICANCE

OIP5-AS1 contributed to EAE development by targeting miR-140-5p/RhoA and activating RhoA/ROCK2 signaling pathway, shedding light on the roles and molecular mechanisms of OIP5-AS1 in the development of MS and providing some candidate targets for the diagnose and treatment of MS.

摘要

目的

多发性硬化症（MS）是全球最常见的神经系统疾病之一。已发现长链非编码RNA OIP5-AS1与MS的病因有关。本研究旨在探讨OIP5-AS1在MS发病过程中的作用及分子机制。

材料与方法

采用RT-qPCR法检测OIP5-AS1、miR-140-5p、IL-17A mRNA和RhoA mRNA的表达。通过流式细胞术测定CD4⁺IL-17细胞比例。采用ELISA法检测IL-17A分泌。通过组织学分析评估细胞炎症浸润和脱髓鞘情况。通过生物信息学分析和荧光素酶报告基因检测验证miR-140-5p与OIP5-AS1或RhoA 3'UTR之间的相互作用。采用蛋白质免疫印迹法检测ROCK2和RhoA的蛋白表达。建立实验性自身免疫性脑脊髓炎（EAE）模型，以探讨OIP5-AS1在MS体内的作用。

主要发现

MS患者中OIP5-AS1表达增强。此外，在辅助性T细胞17（Th17）分化过程中观察到OIP5-AS1水平升高。而且，OIP5-AS1在体外促进Th17分化，并在体内促进EAE的发展。机制研究表明，OIP5-AS1靶向miR-140-5p以调节Th17分化。此外，RhoA是miR-140-5p的靶标，miR-140-5p抑制RhoA/ROCK2信号通路的激活。而且，RhoA上调消除了miR-140-5p对Th17分化的抑制作用。