miR-654-5p Contributes to the Activation and Proliferation of Hepatic Stellate Cells by Targeting RXRα

• Published in: Frontiers in cell and developmental biology Vol. 10; p. 841248
• Main Authors: Ma, Heming, Wang, Xiaomei, Liu, Xu, Wang, Chang, Gao, Xiuzhu, Niu, Junqi
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 06.04.2022
• Subjects: Cell and Developmental Biology; hepatic stellate cells; liver fibrosis; miR-654-5p; RXRα; target; hepatic stellate cells; liver fibrosis; miR-654-5p; RXRα; target
• ISSN: 2296-634X; 2296-634X
• DOI: 10.3389/fcell.2022.841248

Liver fibrosis (LF) is a major disease that threatens human health. Hepatic stellate cells (HSCs) contribute directly to LF via extracellular matrix (ECM) secretion. Moreover, RXRα is an important nuclear receptor that plays a key regulatory role in HSC activation. Meanwhile, microRNAs (miRNAs) have been identified as significant regulators of LF development. In particular, miR-654-5p is involved in cellular migration and proliferation, and bioinformatics analysis, has been identified as a potential factor that targets RXRα in humans and in mice. However, the precise relationship between miR-654-5p and RXRα in the context of LF, remains unknown and is the primary focus of the current study. To establish activated cell model human primary HSCs were cultured and LX-2 cells were stimulated with recombinant human TGF-β1. mRNA and protein levels of RXRα, miR-654-5p and fibrogenic genes were compared in quiescent and activated HSCs. Moreover, after transfected with miR-654-5p mimics, the expression changes of above related genes in LX-2 cells were estimated. Meanwhile, cell proliferation and apoptosis were detected in miR-654-5p overexpressed LX-2 cells. Simultaneously, the targeted binding between miR-654-5p and RXRα was verified in LX-2 cells. Carbon tetrachloride (CCl )-induced mouse model with liver fibrosis was use to research the role of the miR-654-5p . Our results show that miR-654-5p expression levels increased in activated human HSCs and TGFβ-treated LX-2 cells. Moreover, miR-654-5p mimics markedly promoted LX-2 cell proliferation while inhibiting their apoptosis. Accordingly, the expression levels of RXRα are decreased in activated HSCs and LX-2 cells. Additionally, dual-luciferase reporter assay results reveal direct targeting of RXRα by miR-654-5p. Similarly, miR-654-5p overexpression aggravates LF in mice that are intraperitoneally injected with CCl . Taken together, our findings elucidated a novel molecular mechanism with potential use for treatment of LF.