Mechanisms Of S-Palmitoylation Inhibition Reshapes Calcium Homeostasis In Periodontitis

This study investigates whether inhibiting ZDHHC6-mediated S-palmitoylation of calcium transporters rescues ER-mitochondrial dysfunction in chronic periodontitis, thereby restoring immuno-osseous homeostasis ZDHHC6-targeting inhibitors were screened via network pharmacology, molecular dynamics simulations, and docking. In RAW264.7 and mBMSCs subjected to LPS induce with/without inhibitor treatment, ER/mitochondrial stress markers and calcium transporter dynamics were quantified using qRT-PCR, Western blotting, and TEM ultrastructural analysis of mitochondria-associated membranes (MAMs). Functional impacts were assessed via live-cell imaging of Ca²⁺ fluxes (Fluo-4/Rhod-2), ROS generation, and MPTP activation, with proteomic profiling (GO/GSEA) of differentially expressed proteins. Inhibitor-loaded hydrogels were locally administered in murine periodontitis models, with outcomes evaluated by micro-CT bone morphometry, histomorphometry (H&E/Masson/TRAP staining), and immunofluorescence of inflammatory/osteogenic/calcium regulatory proteins. All procedures were approved by the IRB of Chongqing Medical University Stomatological Hospital (2024-145) Computational screening (network pharmacology/MD simulations/docking) identified β-carotene as a natural ZDHHC6 inhibitor targeting calcium transporters.β-carotene treatment rescued LPS-induced ER-mitochondria coupling dysfunction in macrophages and stem cells, ameliorating calcium overload (cytosolic/mitochondrial), ROS overproduction, and MAMs hyperconnectivity via palmitoylation-dependent calcium regulatory axis.Localized β-carotene hydrogel delivery restored periodontal bone volume (micro-CT/histomorphometry), suppressed osteoclastic TRAP⁺ activity while reactivating osteogenesis, achieving immunometabolic-bone axis homeostasis in murine periodontitis models β-carotene inhibits ZDHHC6-mediated calcium transporter palmitoylation, reversing ER-mitochondrial dysfunction to restore immuno-osseous homeostasis in chronic periodontitis