Inhibition of gingival fibroblast necroptosis mediated by RIPK3/MLKL attenuates periodontitis

Abstract Aim Necroptosis participates in the pathogenesis of many inflammatory diseases, including periodontitis. Here, we aimed to investigate the role and mechanism of necroptosis inhibitors in attenuating periodontitis. Materials and Methods The Gene Expression Omnibus (GEO) dataset GSE164241 was re‐analysed to identify the role of necroptosis in periodontitis. Gingival specimens from healthy subjects or periodontitis patients were collected to evaluate the expression level of necroptosis‐associated proteins. The therapeutic effect of necroptosis inhibitors on periodontitis was assessed in vivo and in vitro. Moreover, Transwell assays and Western blotting and siRNA transfection were used to identify the effects of necroptotic human gingival fibroblasts (hGFs) on THP‐1 macrophages. Results Re‐analysis revealed that gingival fibroblasts (GFs) in periodontitis gingiva showed the highest area under the curve score of necroptosis. Elevated levels of necroptosis‐associated proteins were identified in GFs in periodontitis gingiva collected from patients and mice. In ligature‐induced periodontitis mice, local administration of receptor interacting protein kinase 3(RIPK3) inhibitor GSK′872 or sh‐mixed‐lineage kinase domain‐like pseudokinase (Mlkl) markedly abrogated necroptosis and rescued periodontitis. Analogously, necroptosis inhibitors alleviated the inflammatory response and release of damage‐associated molecular patterns in lipopolysaccharide‐ or LAZ (LPS + AZD'5582 + z‐VAD‐fmk, necroptosis inducer)‐induced GFs and then reduced THP‐1 cell migration and M1 polarization. Conclusions Necroptosis in GFs aggravated gingival inflammation and alveolar bone loss. Necroptosis inhibitors attenuate this process by modulating THP‐1 macrophage migration and polarization. This study offers novel insights into the pathogenesis and potential therapeutic targets of periodontitis.