Autophagy upregulates inflammatory cytokines in gingival tissue of patients with periodontitis and lipopolysaccharide-stimulated human gingival fibroblasts.

Abstract

BackgroundPeriodontitis is an inflammatory disease caused by multiple disease‐associated bacterial species in periodontal tissues. Autophagy is known to modulate various inflammation‐driven diseases and inflammatory responses, but the role of autophagy related to the pathogenesis of periodontitis is not fully established. We investigated whether autophagic flux regulated the expression of inflammatory cytokines in the gingiva of periodontitis patients and lipopolysaccharide (LPS)‐stimulated human gingival fibroblasts (HGFs) and the underlying mechanism.MethodsThe mRNA and protein expression of proinflammatory cytokines was assessed in human gingival tissues collected from patients with periodontitis and HGFs treated with LPS. The expression of signaling molecules related to autophagy was evaluated by immunofluorescence and Western blot analyses.ResultsThe expression of interleukin (IL)‐6, tumor necrosis factor‐α (TNF‐α), cyclooxygenase‐2 (COX‐2), and intercellular adhesion molecule‐1 (ICAM‐1) was increased in the gingival tissues of patients with periodontitis. LC3B‐positive cells, a typical autophagic marker, were increased in the gingival tissues of periodontitis patients and LPS‐treated HGFs. The conversion ratio of LC3‐I to LC3‐II was higher in the gingival tissues associated with periodontitis and LPS‐treated HGFs compared to the controls. The autophagy inhibitor 3‐methyladenine (3MA) significantly abrogated the LPS‐sustained inflammatory effect by reducing the expression of IL‐6, TNF‐α, COX‐2, and ICAM‐1 in HGFs. The phosphorylation of protein kinase B (AKT) and protein S6K1 (S6), signals involved in the mTOR‐dependent mechanism, was decreased in gingiva derived from periodontitis patients and LPS‐treated HGFs.ConclusionsAutophagy augmented the production of inflammatory cytokines by mTOR inactivation via the AKT signaling pathway in the gingival tissues of patients with periodontitis and LPS‐stimulated HGFs. These findings would provide a better understanding of the mechanism by which autophagy regulates the inflammatory response associated with periodontal pathogenesis.