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Caspase-3 and gasdermin E mediate macrophage pyroptosis in periodontitis
Xiangru Gao1,2, Shuhan Li1,2, Wenxuan Wang3
1Department of Periodontology, The Affiliated Hospital of Qingdao University, Qingdao, China.
Background And Objectives:
Periodontitis is a chronic inflammatory disease linked to pyroptosis, an inflammatory cell death process. Macrophages are essential for maintaining microenvironment homeostasis, which is crucial for periodontal health. This study explores the mechanisms underlying the relationship between macrophage pyroptosis and periodontitis.
Methods:
Expression of the pyroptosis marker gasdermin E (GSDME) and the macrophage surface marker CD68 was examined by immunofluorescence double staining in healthy and periodontitis gingival tissues. In an in vitro pyroptosis model, RAW264.7 cells were irritated using Porphyromonas gingivalis-lipopolysaccharide (P. gingivalis-LPS) after treatment with either a nuclear factor kappa-B (NF-κB) agonist or inhibitor. The mRNA and protein levels of NF-κB, caspase-3, GSDME, and interleukin-1β (IL-1β) were evaluated through qRT-PCR, western blotting, and ELISA techniques.
Results:
GSDME and CD68 were heavily elevated in inflamed gingival tissues compared to healthy tissues and co-localized in the same region. Furthermore, exposure to P. gingivalis-LPS resulted in a significant upregulation of NF-κB, caspase-3, GSDME, and IL-1β at both the mRNA and protein levels in RAW264.7 cells. NF-κB agonist or inhibitor pretreatment enhanced or inhibited these effects.
Conclusions:
GSDME-mediated macrophage pyroptosis is implicated in periodontitis. Based on in vitro experiments, P. gingivalis-LPS causes pyroptosis in RAW264.7 cells through the caspase-3/GSDME pathway. Furthermore, NF-κB regulates this pyroptotic pathway.
Insights
Periodontitis involves macrophage pyroptosis, a cell death process. This study shows Porphyromonas gingivalis-lipopolysaccharide triggers pyroptosis via the caspase-3/GSDME pathway, regulated by NF-κB.
Area of Science:
- Oral biology and immunology
- Cellular mechanisms of inflammation
- Periodontal disease pathogenesis
Background:
- Periodontitis is a chronic inflammatory disease associated with pyroptosis.
- Macrophages play a critical role in maintaining periodontal homeostasis.
- Understanding macrophage pyroptosis is key to addressing periodontitis.
Purpose of the Study:
- To investigate the role of macrophage pyroptosis in periodontitis.
- To elucidate the molecular mechanisms linking macrophage pyroptosis and periodontitis.
- To explore the involvement of the NF-κB pathway in this process.
Main Methods:
- Immunofluorescence staining for GSDME and CD68 in human gingival tissues.
- In vitro pyroptosis model using RAW264.7 cells stimulated with P. gingivalis-LPS.
- Analysis of NF-κB, caspase-3, GSDME, and IL-1β expression via qRT-PCR, western blotting, and ELISA.
Main Results:
- Elevated GSDME and CD68 expression and co-localization in periodontitis tissues.
- P. gingivalis-LPS significantly upregulated NF-κB, caspase-3, GSDME, and IL-1β in macrophages.
- NF-κB modulation (agonist/inhibitor) affected the pyroptotic pathway.
Conclusions:
- GSDME-mediated macrophage pyroptosis is involved in periodontitis.
- P. gingivalis-LPS induces macrophage pyroptosis through the caspase-3/GSDME pathway.
- The NF-κB pathway regulates P. gingivalis-LPS-induced macrophage pyroptosis.
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