Zhichuanling Oral Liquid Attenuates Airway Inflammation in Asthma by Targeting the STIM1/CHOP/JNK Axis

Ethnopharmacological relevance: Zhichuanling oral liquid (ZCLOL) is a multi‑component traditional Chinese medicine formulation that has been widely used in the clinical management of asthma. However, the mechanisms underlying its attenuation of airway inflammation remain unclear. Aim of the study: This study aimed to elucidate the mechanisms by which ZCLOL mitigates asthma‑related airway inflammation through the integration of in vivo and in vitro models with proteomic profiling. Methods: An ovalbumin (OVA)-induced asthma model was established in BALB/c mice. Lung function, histopathology, serum Th2 cytokines and immunoglobulins, bronchoalveolar lavage fluid (BALF) macrophage phenotypes, lung tissue Ca²⁺ levels, and protein expression were assessed. Data-independent acquisition (DIA) proteomics was performed to identify differentially expressed proteins (DEPs) and enriched pathways. In vitro, an inflammatory BEAS-2B cell model was induced using LPS and IL-13, with BAPTA-AM as a Ca²⁺ chelation control. Intracellular Ca²⁺ levels, ERS stress markers, and expression levels of STIM1/CHOP/JNK-related proteins were evaluated. Results: ZCLOL significantly improved airway hyperresponsiveness and reduced airway inflammation in OVA-induced mice, as evidenced by decreased specific airway resistance (sRaw), alleviated histopathological damage, and reduced inflammatory cell infiltration. Serum levels of OVA-sIgG, IgE, IL-4, IL-5, and IL-13, as well as BALF M1 macrophage activation, were markedly reduced. DIA proteomics revealed significant enrichment of the calcium signaling pathway among DEPs. ZCLOL downregulated lung tissue expression of STIM1, Bip, Ero1-Lɑ, PDI, CHOP, and p-JNK, accompanied by reduced Ca²⁺ accumulation. In vitro, ZCLOL suppressed Ca²⁺ overload and endoplasmic reticulum stress (ERS), and inhibited activation of the STIM1/CHOP/JNK axis, with additive effects observed when combined with BAPTA-AM. Conclusion: ZCLOL alleviates airway inflammation and hyperresponsiveness in asthma by restoring Ca²⁺ homeostasis and suppressing ERS through inhibition of the STIM1/CHOP/JNK axis. These findings provide mechanistic evidence supporting ZCLOL as a potential multi-target therapeutic strategy for asthma.