ELAVL1 modulates periodontal ligament fibroblast pyroptosis in periodontitis through the JAK2/STAT3/NLRP3 axis

Abstract Periodontitis is an inflammatory disorder that leads to the destruction of periodontal tissues. Pyroptosis, related to the Nod-like receptor protein 3 (NLRP3) inflammasome, is implicated in the pathogenesis of periodontitis. ELAVL1 (embryonic lethal, abnormal vision, Drosophila-like 1), an N6-methyladenine (m6A) reader, is associated with inflammatory responses; however, its role in periodontitis requires further clarification. Our research elucidated the regulatory functions of ELAVL1 in periodontitis and explored the underlying mechanisms. Expression of ELAVL1 in periodontitis mice and Porphyromonas gingivalis lipopolysaccharide (P.g-LPS)-induced eriodontal ligament fibroblasts (PLFs) was assessed via quantitative real-time PCR (qRT-PCR) and western blot. Enzyme-linked immunosorbent assay (ELISA) was performed to analyse IL-1β and IL-18 levels. The associated protein levels of the NLRP3 inflammasome, bone formation, and the JAK2/STAT3 (Janus kinase 2 / signal transducer and activator of transcription 3) pathway were detected using western blot. Pyroptosis was evaluated by flow cytometry. RNA immunoprecipitation (RIP), methylated RIP (MeRIP), and dual-luciferase reporter assay were performed to validate the association between ELAVL1 and JAK2 mRNA. Immunofluorescence was used to detect ELAVL1, NLRP3, and Vimentin expression in periodontal tissues. The pathological changes in periodontal tissues were determined by micro-CT and hematoxylin and eosin (H&E) staining. We found that ELAVL1 was upregulated in periodontitis tissues and P.g-LPS-stimulated PLFs. Mechanistically, ELAVL1 binds to JAK2 mRNA, enhancing its m6A modification and stability, thereby activating the JAK2/STAT3 signaling. Knockdown of ELAVL1 decreased NLRP3 inflammasome activation and pyroptosis in PLFs, inhibited inflammatory factor secretion, and inhibited activation of the JAK2/STAT3 pathway. However, these effects were partially reversed following treatment with C-A1. Furthermore, ELAVL1 knockdown attenuated alveolar bone loss, decreased p-JAK2 and p-STAT3 levels, and suppressed NLRP3-mediated pyroptosis in vivo. We conclude that ELAVL1 promotes periodontitis by mediating m6A modification of JAK2 and activating the JAK2/STAT3/NLRP3-mediated pyroptosis, suggesting its promise as a novel treatment target for periodontitis.