降气平哮方调控CLCA1/Ca2+/SNARE信号改善哮喘气道黏液高分泌的机制研究OA

OBJECTIVE To investigate the mechanism by which Jiangqi Pingxiao Formula(JQPXF)regulates Ca2+and mem-brane fusion to attenuate airway mucus hypersecretion in asthma.METHODS An ovalbumin-induced asthma model was established in mice.The experimental groups included the normal control group,the asthma model group,JQPXF low-(10.0 g·kg⁻¹),medium-(20.0 g·kg⁻¹),and high-dose(30.0 g·kg⁻¹)groups,the dexamethasone group(1.0 mg·kg⁻¹),and the ambroxol hydrochloride group(8.0 mg·kg⁻¹).During the model construction period,the status of mice were observed through behavioral observation,and lung patho-logical status were evaluated by HE and PAS staining.The levels of inflammatory cytokines interleukin(IL)-4 and IL-13 in mouse lungs were detected using the ELISA method.qPCR was used to detect mucin mRNA expression,calcium ion(Ca²⁺)concentration in mouse lung tissue was measured by colorimetric assays,and immunohistochemistry and Western blot analyses were used to assess the expression of calcium-activated chloride channel modulator 1(CLCA1)and membrane fusion-related proteins.RESULTS Com-pared with the normal group,the model group exhibited significantly increased airway inflammation scores,mucus secretion,IL-13 and IL-4 levels,calcium ion concentration,and expression of CLCA1;synaptophysin-associated protein 23(SNAP23),soluble N-ethylmaleimide-sensitive receptor protein(SNARE),synaptotagmin(STX3),vesicle-associated membrane protein 8(VAMP8),and vesicle-associated membrane protein 2(VAMP2)were significantly elevated(P<0.05,P<0.01,P<0.001).Compared with the model group,JQPXF effectively improved various clinical signs in asthmatic mice,significantly alleviating airway inflammation and mucus se-cretion(P<0.05,P<0.001);reduced IL-4 and IL-13 levels in lung tissue(P<0.001);suppressed the overexpression of mucin genes(P<0.05,P<0.01,P<0.001);decreased intracellular Ca²⁺ concentration(P<0.05,P<0.001);and downregulated the expression of key proteins in the membrane fusion pathway,including CLCA1,SNAP23,SNARE,STX3,VAMP8 and VAMP2.CONCLUSION JQPXF may ameliorate mucus hypersecretion in asthmatic airways by suppressing the CLCA1/Ca²⁺/SNARE signaling pathway,provid-ing a novel molecular mechanism target for JQPXF therapy.