Sangma Zhike Formula alleviates airway inflammation and hyperresponsiveness in rats with postinfectious cough by inhibiting the TRPV1-SP/CGRP and pyroptosis pathways

doi:10.12122/j.issn.1673-4254.2025.09.03

Abstract

Abstract:

Objective To investigate the therapeutic mechanism of Sangma Zhike Formula (SMZKF) for relieving cough sensitivity and airway inflammation in rats with postinfectious cough (PIC). Methods Male SD rat models were established by cigarette smoke exposure with intranasal LPS instillation and capsaicin aerosol inhalation. From day 19 following the start of PIC modeling, the rats received daily treatment with saline (model group), low-, medium-, and high-dose SMZKF, and compound methoxyphenamine (ASM) via gavage for 10 consecutive days (n=8). The assessments included behavioral changes, cough sensitivity (latency and frequency), lung histopathology, inflammatory cell counts and cytokine/mediator levels in the bronchoalveolar lavage fluid (BALF), oxidative stress markers in the lung tissue, and expressions of proteins related with cough hypersensitivity and pyroptosis. Results The rat models of PIC exhibited reduced mental alertness, accelerated respiration, and pronounced symptoms such as coughing, sneezing, and facial scratching with significantly shortened cough latency and increased 5-min cough frequency. Histopathological analysis revealed collapsed alveolar structures, thickened alveolar septa, and extensive inflammatory cell infiltration in the bronchi and peribronchial regions, accompanied by elevated bronchial and alveolar inflammation scores of the rat models. In the BALF, inflammatory cell counts and the levels of IL-1β, TNF-α, IL-6, COX-2, PGE-2, and TXA-2 were all markedly elevated, and the pulmonary oxidative stress markers (ROS and MDA) and myeloperoxidase (MPO) activity were also significantly increased. The pulmonary expressions of cough hypersensitivity-related proteins (TRPV1, SP, CGRP, and NK1R) and pyroptosis-associated markers (P-NF-κB, NLRP3, ACS, cleaved caspase-1, cleaved IL-1β, and GSDMD-N) were significantly upregulated in the model group. SMZKF interventions significantly ameliorated these pathological changes in the rat models, and high-dose SMZKF produced a similar therapeutic efficacy to that of ASM. Conclusion SMZKF alleviates cough sensitivity and airway inflammation in PIC rats possibly by inhibiting TRPV1-mediated SP/NK1R signaling and the NLRP3/caspase-1/GSDMD pyroptosis pathway.

Key words: postinfectious cough, Sangma Zhike Formula, cough sensitivity, airway inflammation, mechanism

Qinjun YANG, Hongyu ZHU, Yuan GAO, Cheng YANG, Tong LIU, Lu ZHANG, Jiabing TONG, Zegeng LI. Sangma Zhike Formula alleviates airway inflammation and hyperresponsiveness in rats with postinfectious cough by inhibiting the TRPV1-SP/CGRP and pyroptosis pathways[J]. Journal of Southern Medical University, 2025, 45(9): 1830-1839.

Figures/Tables 7

Fig.1 Schematic diagram of postinfectious cough (PIC) modeling in rats and drug administration.

Fig.2 Cough latency (A) and number of coughs within 5 min (B) in each group. *P<0.05 vs CON group, #P<0.05 vs PIC group.

Fig.3 Lung tissue pathology and bronchial and alveolar inflammation scores in each group of rats. A: HE staining of rat lung tissue (Original magnification: ×400). B: Bronchial inflammation scores. C: Alveolar inflammation scores. *P<0.05 vs CON group, #P<0.05 vs PIC group.

Fig.4 Inflammatory cell counts in BALF of the rats in each group. A: WBC. B: LYM. C: NEU. D: MON. E: EOS. *P<0.05 vs CON group, #P<0.05 vs PIC group.

Fig.5 Levels of inflammatory factors and mediators in the BALF of the rats in each group. A: IL-1β. B: TNF-α. C: IL-6. D: PGE-2. E: COX-2. F: TXA-2. *P<0.05 vs CON group, #P<0.05 vs PIC group.

Fig.6 Levels of ROS and MDA in the lung tissues of the rats in each group. A: Flow cytometry for detecting ROS in the lung tissue. B: ROS. C: MDA. D: MPO. *P<0.05 vs CON group, #P<0.05 vs PIC group.

Fig.7 Expressions of airway sensitivity-related proteins in the lung tissues of the rats in each group. A: Western blotting for detecting expressions of TRPV1, SP, CGRP and NK1R in the lung tissue. B: TRPV1. C: SP. D: CGRP. E: NK1R. *P<0.05 vs CON group, #P<0.05 vs PIC group.

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