Mitophagy Attenuates Pyroptosis in Human Gingival Fibroblasts through Inhibition of NLRP3 Inflammasome Activation

Mitophagy Attenuates Pyroptosis in Human Gingival Fibroblasts through Inhibition of NLRP3 Inflammasome Activation | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 5 March 2025 V1 Latest version Share on Mitophagy Attenuates Pyroptosis in Human Gingival Fibroblasts through Inhibition of NLRP3 Inflammasome Activation Authors : Chun-Sheng Bi 0009-0003-5955-9319 , Yi-Kuan Wu , Hao-Yu Li , Zi-Jian Cheng , and Xiao-Jun Li [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.174115997.75468747/v1 Published Archives of Oral Biology Version of record Peer review timeline 192 views 153 downloads Contents Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Objectives: The NLRP3 inflammasome-mediated pyroptosis in gingival fibroblasts (GFs) has been implicated in the pathogenesis of periodontitis. Mitochondrial dysfunction represents a critical upstream event in NLRP3 inflammasome activation, while mitophagy functions as an essential cellular mechanism for maintaining mitochondrial homeostasis. Therefore, the present study was designed to elucidate the regulatory role and molecular mechanisms of mitophagy in modulating pyroptosis in GFs. Materials and methods: Human GFs were used in this study. An in vitro inflammatory environment was established using 5 μg/ml lipopolysaccharide (LPS). GFs exposed to LPS were treated with 10 μM P62-mediated mitophagy inducer (PMI), while healthy GFs not exposed to LPS received 10 μM Mdivi-1 intervention. Mitophagy was visualized by immunofluorescence, and the expression levels of PINK1, Parkin, and Beclin-1 were assessed via qRT-PCR and Western blot. Mitochondrial morphology was examined by Transmission electron microscopy (TEM), and mitochondrial damage was evaluated through measurements of mitochondrial membrane potential (MMP) and intracellular reactive oxygen species (ROS) and mitochondrial ROS (mtROS) levels using flow cytometry. NLRP3 inflammasome activation was analyzed by qRT-PCR and Western blot. Propidium iodide (PI) staining, lactate dehydrogenase (LDH) release, IL-1β and IL-18 expression levels and apoptosis assays were also detected to assess pyroptosis in GFs. Results: LPS stimulation suppressed mitophagy in GFs, which was reversed by mitophagy activator. In contrast, mitophagy inhibition decreased basal mitophagy in healthy GFs. Mitophagy activation maintained mitochondrial function by enhancing MMP and reducing ROS/mtROS accumulation and mtDNA/nDNA ratio in LPS-stimulated GFs. Furthermore, mitophagy attenuated LPS-induced NLRP3 inflammasome activation and decreased GF death, and IL-1β/IL-18 production and secretion. Conclusions: Mitophagy alleviates NLRP3 inflammasome-dependent pyroptosis of GFs by maintaining mitochondrial homeostasis. Supplementary Material File (main document.docx) Download 1.08 MB Information & Authors Information Version history V1 Version 1 05 March 2025 Peer review timeline Published Archives of Oral Biology Version of Record 1 Nov 2025 Published Copyright This work is licensed under a Non Exclusive No Reuse License. Keywords gingival fibroblasts mitophagy nlrp3 inflammasome periodontitis pyroptosis Authors Affiliations Chun-Sheng Bi 0009-0003-5955-9319 Zhejiang University School of Medicine Stomatology Hospital View all articles by this author Yi-Kuan Wu Zhejiang University School of Medicine Stomatology Hospital View all articles by this author Hao-Yu Li Zhejiang University School of Medicine Stomatology Hospital View all articles by this author Zi-Jian Cheng Zhejiang University School of Medicine Stomatology Hospital View all articles by this author Xiao-Jun Li [email protected] Zhejiang University School of Medicine Stomatology Hospital View all articles by this author Metrics & Citations Metrics Article Usage 192 views 153 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Chun-Sheng Bi, Yi-Kuan Wu, Hao-Yu Li, et al. Mitophagy Attenuates Pyroptosis in Human Gingival Fibroblasts through Inhibition of NLRP3 Inflammasome Activation. Authorea . 05 March 2025. DOI: https://doi.org/10.22541/au.174115997.75468747/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. For more information or tips please see 'Downloading to a citation manager' in the Help menu . Format Please select one from the list RIS (ProCite, Reference Manager) EndNote BibTex Medlars RefWorks Direct import Tips for downloading citations document.getElementById('citMgrHelpLink').addEventListener('click', function() { popupHelp(this.href); return false; }); $(".js__slcInclude").on("change", function(e){ if ($(this).val() == 'refworks') $('#direct').prop("checked", false); $('#direct').prop("disabled", ($(this).val() == 'refworks')); }); View Options View options PDF View PDF Figures Tables Media Share Share Share article link Copy Link Copied! Copying failed. 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