Thiolutin Reverses Lipopolysaccharide/Adenosine Triphosphate-Induced Pyroptosis and Inflammation in Human Gingival Fibroblasts by Suppressing NLRP3 Inflammasome

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<p>Pharmacological studies revealed that thiolutin (THL) prevented or treated NLRP3-associated inflammatory diseases. The activation of the NLRP3 inflammasome plays a crucial role in the pathogenesis of periodontitis. However, no study has demonstrated the therapeutic potential of THL in periodontitis. Human gingival fibroblasts (HGFs) were incubated with 1 μg/mL of lipopolysaccharide (LPS) and 5 mM of adenosine triphosphate (ATP), activating the NLRP3-mediated inflammatory response and intervened with THL at doses of 10 nM. Subsequently, cell viability, lactate dehydrogenase release, cell apoptosis, the contents of proinflammatory and oxidative stress factors, including superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), reactive oxygen species (ROS), interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-1β, and IL-18, and the expression levels of NOD-like receptor protein 3 (NLRP3)-related proteins were detected by CCK-8, propidium iodide fluorescence staining, enzyme-linked immunosorbent assay, quantitative reverse transcription-polymerase chain reaction, and western blotting, respectively. THL countered the inhibitory effects on cell viability and promoted the effects on cell apoptosis, production of IL-6, TNF-α, IL-1β, and IL-18, and NLRP3-related proteins induced by LPS/ATP stimulation. Furthermore, the SOD, GSH, ROS, and MDA levels were not affected by THL intervention, suggesting that the recovery of LPS/ATP-induced HGF injury by THL depends on inflammatory response but not oxidative stress. The present study demonstrated that THL repressed the release of inflammatory factors mediated by NLRP3 inflammasome activation in LPS/ATP-induced HGFs rather than oxidative stress levels to ameliorate inflammatory injury and pyroptosis and exerted protective effects on HGFs.</p>

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