Mechanism study of Kong Xian-Dan's improvement of asthma via the PLC/IP3R/Ca2+/TMEM16A pathway based on network pharmacology and LC-MS/MS

s Background Kongxian Dan (KXD), a classic‌ traditional Chinese prescription for treatingexcessive mucus secretion, which possesses broad therapeutic potential. This study aims to identify its active components against asthma and elucidate the underlying mechanisms. Methods Active components of KXD absorbed into mouse serum were identified using LC-MS/MS. Potential targets and related signaling pathways were predicted via network pharmacology. An OVA-induced mouse asthma model as well as human bronchial epithelial BEAS-2B cells were established to evaluate the therapeutic effects. Inflammatory cells, cytokines and oxidative stress markers in BALF and lung tissue were measured. Histopathology was performed to assess airway inflammation, mucus secretion, remodeling, and hepatorenal toxicity. The expression of mucin (MUC5AC) and TMEM16A was detected via immunofluorescence, and related key proteins were analyzed by western blot. Results Sixteen active components derived from KXD were identified. Network pharmacology analysis suggested a critical association with calcium signaling pathways. In vivo and in vitro experiments showed that KXD significantly alleviated airway inflammation, reduced inflammatory cell infiltration and cytokine levels, and suppressed decreased oxidative stress. It also attenuated mucus hypersecretion and airway remodeling without hepatorenal toxicity. Mechanistically, KXD reduced intracellular calcium concentration by downregulating PLC, IP3R, and TMEM16A expression. Conclusion KXD mitigates asthma-related airway inflammation and excessive mucus secretion, potentially through the inhibition of the PLC/IP3R/Ca²⁺/TMEM16A signaling pathway.