Dual inhibition of caspase-11 pyroptosis and NLRP3 inflammasome by Huanglian Jiedu Decoction: A polypharmacological strategy for sepsis management.

BACKGROUND

Sepsis, responsible for 19.7% of global deaths, is characterized by caspase-11-mediated pyroptosis triggered by lipopolysaccharide (LPS). Huanglian Jiedu Decoction (HLJDD), a traditional Chinese formula comprising Coptis chinensis and related herbs, has demonstrated anti-inflammatory efficacy, yet its molecular targets in caspase-11 regulation remain undefined.

PURPOSE: This study aims to mechanistically decode HLJDD's multi-target effects on caspase-11 signaling and pyroptosis pathways, bridging traditional detoxification principles with modern inflammasome biology.

METHODS

Three integrated strategies were employed: (1) Pharmacokinetic profiling via LC-MS/MS and MALDI-MSI identified bioactive components and organ-specific distribution; (2) Qualitative analysis of constituents using UHPLC-QTOF-MS; (3) Biolayer interferometry and 100-ns molecular dynamics simulations quantified caspase-11 binding; (4) LPS-induced septic mice (12 mg/kg) received HLJDD (1.048-4.192 g/kg) for cytokine profiling (ELISA), histopathology, and immunoblotting of NLRP3/ASC/caspase-1 pathways. Network pharmacology analyzed 1962 shared targets.

RESULTS

HLJDD exhibited dose-dependent efficacy: 33.8% IL-1β reduction, 41.2% IL-18 suppression (p<0.001), and complete GSDMD cleavage inhibition. Berberine dominated caspase-11 binding via Tyr208/Leu209 interactions, while geniposide modulated HMGB1 release (43.8% downregulation). Concurrent NLRP3-ASC complex disassembly (31.8% reduction) and hepatic protection (edema score: 4.2→1.7) confirmed multi-organ synergy.

CONCLUSION

HLJDD's therapeutic paradigm operates through dual mechanisms: direct caspase-11 inhibition and NLRP3 inflammasome disruption, achieving synchronized blockade of pyroptosis initiation and cytokine amplification. The formula's phytochemical consortium (berberine-baicalin-geniposide) exemplifies functional complementarity, outperforming single-target agents. These findings validate TCM's potential in precision sepsis management, providing molecular evidence for the Traditional Chinese Medicine concept of "clearing heat and detoxifying."