Role of IL-10 in regulating macrophage pyroptosis in immune thrombocytopenia

Abstract

Abstract Introduction Primary immune thrombocytopenia (ITP) is the most prevalent hemorrhagic disorder in clinical practice, with relapse and refractoriness representing urgent therapeutic challenges. Macrophage pyroptosis-driven inflammatory and immune dysregulation serves as a pivotal mechanism in the pathogenesis of autoimmune diseases and constitutes a novel therapeutic target for inflammatory disorders. Previous studies demonstrated that IL-10 gene knockout mice exhibit significantly elevated expression of key pyroptotic proteins in macrophages, indicating a correlation between IL-10 and macrophage pyroptosis. We hypothesize that ITP features both excessive macrophage pyroptosis and IL-10 deficiency, and that exogenous IL-10 supplementation may exert therapeutic effects by suppressing the NLRP3-mediated canonical pyroptosis pathway. Methods 1.Collection of clinical samples from ITP patients Peripheral blood and bone marrow samples from ITP patients were collected. The expression levels of key molecules in the canonical pyroptosis pathway (NLRP3, activated caspase-1, and GSDMD-NT) were detected using flow cytometry and Western blotting. IL-10 levels were quantified concurrently. 2.Induction and intervention of pyroptosis models A pyroptosis model was established using THP-1 cells stimulated with either plasma from ITP patients or Nigericin + LPS. Concurrently with pyroptosis induction, IL-10 therapeutic intervention was administered. The expression profiles of NLRP3, activated caspase-1, and GSDMD-NT in the canonical pyroptosis pathway were systematically assessed. Results 1.Clinical findings in ITP patients Macrophages derived from ITP patients show excessive activation of the NLRP3/caspase-1/GSDMD-NT-dependent pathway, accompanied by significantly elevated plasma levels of pyroptosis-related cytokines IL-1β and IL-18. Compared with healthy controls, IL-10 levels are substantially downregulated; notably, steroid-dependent or steroid-resistant patients exhibit a further reduction in IL-10 compared to newly diagnosed ITP cases. 2.In vitro experimental validation Plasma from ITP patients induces hyperactivation of the NLRP3/caspase-1/GSDMD-NT-dependent pathway in macrophages. However, exogenous supplementation with IL-10 significantly suppresses the activation of these canonical pyroptosis executors. Consistently, in LPS + Nigericin-induced positive control models of pyroptosis, administration of IL-10 markedly attenuates the progression of pyroptosis in macrophages. Conclusion Excessive macrophage pyroptosis and deficient IL-10 expression are characteristic features of ITP. IL-10 potentially attenuates ITP progression by suppressing NLRP3/caspase-1/GSDMD-NT-mediated canonical pyroptosis pathway activation in macrophages.