lncRNA XIST inhibition promotes M2 polarization of microglial and aggravates the spinal cord injury via regulating miR-124-3p / IRF1 axis

Jin Yang; Zhiqiang Gong; Junjie Dong; Hangchuan Bi; Bing Wang; Kaili Du; Chunqiang Zhang; Lingqiang Chen

doi:10.1016/j.heliyon.2023.e17852

lncRNA XIST inhibition promotes M2 polarization of microglial and aggravates the spinal cord injury via regulating miR-124-3p / IRF1 axis

Heliyon. 2023 Jul 3;9(7):e17852. doi: 10.1016/j.heliyon.2023.e17852. eCollection 2023 Jul.

Authors

Jin Yang¹, Zhiqiang Gong¹, Junjie Dong¹, Hangchuan Bi¹, Bing Wang¹, Kaili Du¹, Chunqiang Zhang¹, Lingqiang Chen¹

Affiliation

¹ Department of Orthopaedics, First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan, China.

Abstract

Spinal cord injury (SCI) has a high disability rate and mortality rate. Recently, LncRNA XIST has been found to be involved in the regulation of inflammatory responses. Therefore, we aimed to investigate the role of XIST in the occurrence and development of SCI and the specific regulation mechanism. Methods: 100 ng/mL lipopolysaccharide (LPS) was used to treat mouse microglia BV2 cells. Hitting spinal cord was performed to C57BL/6 mice for establishing SCI model. Real-time reverse transcriptase-polymerase chain reaction (RT-qPCR), Western blot, Immunofluorescence (IF) and Enzyme linked immunosorbent assay (ELISA) experiments were used to explore the function of XIST, miR-124-3p and IRF1 in LPS-induced BV2 cells. RT-qPCR, Nissl staining, IF, Western blot and ELISA experiment were performed to study the function of XIST in SCI mice. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP), RT-qPCR and Western blot assays were utilized to identify the interaction among XIST, miR-124-3p and IRF1. Results: XIST was upregulated in LPS-induced BV2 cells and spinal cord tissues of SCI mice. Overexpression of XIST promoted the M1 microphages polarization and cytokines concentration in LPS-stimulated BV2 cells, aggravated SCI of mice. Downregulated XIST promoted M1-to-M2 conversion of microglial and relieved the injury of SCI mice. Mechanism verification indicated that XIST acted as a molecular sponge of miR-124-3p and regulated IRF1 expression. Increased miR-124-3p or reduced IRF1 inhibited M1 polarization of microglial and decreased the production of inflammatory cytokines in LPS-induced BV2 cells. Increased XIST or decreased miR-124-3p had an opposite of on LPS-induced BV2 cells. Conclusion: Overexpression of XIST enhanced M1 polarization of microglia and promoted the level of inflammatory cytokines through sponging miR-124-3p and regulating IRF1 expression.

Keywords: IRF1; Microglia; Polarization; Spinal cord injury; lncRNA XIST; miR-124–3p.