High glucose promotes lipopolysaccharide‐induced macrophage pyroptosis through GSDME O‐GlcNAcylation

Abstract Aim The high glucose (HG) environment in diabetic periodontitis aggravates the damage of periodontal tissue. Pyroptosis has been shown to be positively correlated with the severity of periodontitis, including macrophage pyroptosis. O‐GlcNAcylation is a posttranslational modification that is involved in the pathogenesis of periodontitis. However, whether HG regulates macrophage pyroptosis through O‐GlcNAcylation remains uncertain. This study aimed to investigate the effect of HG on the O‐GlcNAcylation level of a pyroptosis regulator GSDME in macrophages to further probe the mechanisms of diabetic periodontitis. Methods Blood samples were collected from patients with diabetic periodontitis. THP‐1 monocytes were induced to differentiate into macrophages by phorbol 12‐myristate 13‐acetate and then treated with HG to simulate periodontitis in vitro. GSDME expression of blood samples and macrophages was measured by quantitative real‐time PCR. Pyroptosis was assessed by propidium iodide staining, measurement of cell viability, cytotoxicity, protein levels of inflammation factors, and pyroptosis‐related proteins. O‐GlcNAcylation of GSDME was analyzed using co‐immunoprecipitation (co‐IP), IP, and western blot. Results The results showed that GSDME expression was elevated in patients with periodontitis and HG‐treated macrophages. HG inhibited cell viability but increased LDH content, levels of IL‐1β, IL‐18, TNF‐α, NLRP3, GSDMD, and Caspase‐1, indicating that HG promoted pyroptosis of macrophages, which was reversed by GSDME knockdown. HG treatment increased O‐GlcNAcylation in macrophages. Mechanically, GSDME interacted with OGT, and OGT knockdown suppressed O‐GlcNAcylation of GSDME at Ser (S)339 site. Knockdown of OGT inhibited pyroptosis in HG‐treated macrophages, while GSDME overexpression partially reversed this inhibition. Conclusion HG treatment enhanced OGT‐mediated GSDME O‐GlcNAcylation, thereby augmenting pyroptosis in LPS‐induced macrophages. These results may provide a novel sight for the treatment of periodontitis.