| Objective To investigate the protective mechanism of the bone regeneration pills (BRP) against hormone-induced pyroptosis of bone marrow mesenchymal stem cells (BMSCs) and the impact on microtubule-mediated NLRP3 inflammasome assembly. Methods BMSCs were divided into blank group, model group, model group + paclitaxel group, model group + nocardazole group, and model group + BRP serum group. Serum intervention BMSCs containing BRP were prepared. Cell viability was assessed using CCK-8 assay, apoptosis rate was determined by flow cytometry, and WB was used to detect the expressions of AC-α-tubulin, NLRP3, Caspase-1 and GSDMD. The expression of ASC in cells and co-localized expression of α-tubulin and NLRP3 were detected with immunofluorescence. ELISA was used to detect the levels of IL-1β and IL-18. Results Compared to the blank group, the model group exhibited reduced cell viability, increased apoptosis levels, upregulated expression of AC-α-tubulin, NLRP3, Caspase-1, and GSDMD proteins, increased fluorescence intensity of ASC inflammatory specks, and enhanced co-localization of α-tubulin and NLRP3 (P<0.01). In comparison to the model group, the paclitaxel group displayed decreased cell viability, increased apoptosis levels, upregulated expression of AC-α-tubulin, NLRP3, Caspase-1 and GSDMD (P<0.01), increased fluorescence intensity of ASC inflammatory specks (P<0.05), and enhanced co-localization of α-tubulin and NLRP3 (P<0.01). IL-1β and IL-18 contents increased (P<0.01, P<0.05). Both the nocardazole and BRP serum groups showed increased cell viability, decreased apoptosis levels, downregulated expressions of AC-α-tubulin, NLRP3, Caspase-1, and GSDMD proteins (P<0.01), decreased fluorescence intensity of ASC inflammatory specks (P<0.05), and reduced co-localization of α-tubulin and NLRP3 (P<0.01). IL-1β and IL-18 contents decreased (P<0.01, P<0.05). Conclusion BRP inhibits α-tubulin acetylation, disrupts microtubule structure, suppresses NLRP3 inflammasome assembly, and reduces BMSC pyroptosis. |