Background: Chronic periodontitis, one of the most prevalent oral diseases, is associated with Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS) infection and has profound effects on type 2 diabetes mellitus (t2DM). Metformin, a well-known antidiabetic agent, has been reported to exert anti-inflammatory effects on various cells. This study aims to investigate the role of metformin on LPS-influenced inflammatory response in human gingival fibroblasts (HGFs).
Methods: Dose-dependent additive effects of metformin on LPS-influenced HGFs were detected. Cell-counting assay was used to determine effects of metformin and LPS on viability of HGFs. Enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction (qRT-PCR) were applied to detect levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α in differently treated cells. Activating transcription factor-3 (ATF3) small interfering (si)RNA transfection was used to determine the mechanism of metformin action, and the transfection efficiency was observed by fluorescence microscope. Effects of ATF3 knockdown were determined by qRT-PCR and Western blot.
Results: Results showed that 5 μg/mL Pg LPS and 0.1, 0.5, and 1 mM metformin exhibited no toxicity to HGFs, and metformin inhibited LPS-influenced IL-1β, IL-6, and TNF-α production in a dose-dependent manner. Metformin and LPS could synergistically facilitate ATF3 expression, and ATF3 knockdown abolished inhibitory effects of metformin on LPS-influenced inflammatory cytokine production in HGFs.
Conclusion: The present study confirms that metformin suppresses LPS-enhanced IL-6, IL-1β, and TNF-α production in HGFs via increasing ATF3 expression.
Keywords: Activating transcription factor 3; anti-inflammatory agents; cytokines; diabetes mellitus; fibroblasts; periodontitis.