Abstract
<title>Abstract</title> <p> <bold>Background:</bold> Cytokine storm (CS) is a life-threatening hyperinflammatory syndrome. Although andrographolide sulfonate (Andro-S) is clinically used for respiratory inflammation, its tissue distribution and molecular mechanisms in CS remain insufficiently defined. This study aims to investigate the protective mechanism of Andro-S against LPS-induced CS. <bold>Methods:</bold> In a murine model, the efficacy of Andro-S was evaluated. A self-developed 384 In situ Tissue Acquisition Device (384 IsTad) was coupled with MALDI mass spectrometry imaging to visualize Andro-S distribution within pulmonary lesions. An integrated spatial pharmacoproteomic strategy, molecular docking, and cellular thermal shift assays (CETSA) were applied to identify targets and inflammatory pathways, supported by statistical analysis. <bold>Results:</bold> Andro-S significantly improved survival, attenuated lung and spleen injuries, and reduced systemic overproduction of TNF-α, IL-1β, and IL-6. Proteomic and target validation identified IFIT2 as a key candidate target, through which Andro-S suppressed TLR4/Caspase/GSDMD-associated pyroptotic signaling. Notably, chemically distinct components of Andro-S exhibited differential target preferences, demonstrating molecular complementarity. <bold>Conclusion:</bold> Andro-S alleviates CS by restoring IFIT2-associated immune regulation and limiting Caspase-1/GSDMD-dependent pyroptosis. This study establishes a spatially resolved framework for investigating complex botanical derivatives. </p>