Mechanism of lncRNA ZFAS1 mediating nucleus pulposus cell pyroptosis in intervertebral disc degeneration

Abstract

Background: This study investigates the mechanism of lncRNA ZFAS1 in pyroptosis of TNF-α-induced nucleus pulposus cells (NPCs) in intervertebral disc degeneration (IDD). Methods: Mouse NPCs were isolated and induced by TNF-α to establish a cell model of IDD. The cell viability was evaluated by MTT assay. NLRP3, GSDMD-N, and cleaved-Caspase1 expressions were detected by Western blot. IL-1β and IL-18 contents were detected by ELISA. RT-qPCR was performed to determine ZFAS1, miR-155-3p, and METTL14 expressions. After intervening in ZFAS1 expression, the effect of ZFAS1 on pyroptosis was verified by Western blot and ELISA assays. RNA pull down or dual luciferase assay verified the binding between ZFAS1, miR-155-3p, and METTL14. Results: TNF-α induced pyroptosis of NPCs and promoted ZFAS1 expression. Silence of ZFAS1 repressed pyroptosis of TNF-α-induced NPCs. Mechanistically, ZFAS1 upregulated the transcription of METTL14 by competitively binding to miR-155-3p, thus enhancing NLRP3/Caspase-1-mediated NPC pyroptosis. Inhibition of miR-155-3p or overexpression of METTL14 alleviated the inhibitory effect of ZFAS1 silencing on TNF-α-treated NPC pyroptosis. Conclusion: ZFAS1 facilitates NLRP3/Caspase-1-mediated pyroptosis of NPCs in IDD via miR-155-3p/METTL14 axis.