Huan Li,1, Lingxue Jiang,1, Jing Qian,2, Wenping Bao,3 Kenan Huang,4 Dongyun Li,1 Aihua Zhang,5 Jianguo Sun,6 Jialong Lu,1 Danxia Wei1 1Department of Nephrology and Rheumatology, The Third Affiliated Hospital of Yunnan University of Traditional Chinese Medicine, Kunming, Yunnan, Peopleâs Republic of China; 2Department of Rehabilitation, Second Affliated Hospital of Naval Medical University, Shanghai, Peopleâs Republic of China; 3Department of Traditional Chinese Medicine, Dounan Community Health Service Center of Chenggong District, Kunming, Yunnan, Peopleâs Republic of China; 4Department of Thoracic Surgery, Second Affliated Hospital of Naval Medical University, Shanghai, Peopleâs Republic of China; 5Department of Pulmonary Medicine, The Third Affiliated Hospital of Yunnan University of Traditional Chinese Medicine, Kunming, Yunnan, Peopleâs Republic of China; 6Department of Pharmacy, Second Affliated Hospital of Naval Medical University, Shanghai, Peopleâs Republic of ChinaThese authors have contributed equally to this workCorrespondence: Jianguo Sun, Department of Pharmacy, Second Affiliated Hospital of Naval Medical University, No. 415, Fengyang Road, Shanghai, 200003, Peopleâs Republic of China, Tel +86-021-81886181, Email 1240603832@stu.cqmu.edu.cn Danxia Wei, Department of Nephrology and Rheumatology, The Third Affiliated Hospital of Yunnan University of Traditional Chinese Medicine, No. 2628 Xiangyuan Street, Kunming, Yunnan, 650500, Peopleâs Republic of China, Tel +86-0871-63125335, Email 15398558606@163.comPurpose: The in-house preparation Qingre Runzao formula (QRRZF) has been used for decades in our hospital to treat asthma with good efficacy, but its underlying mechanisms and key components remain unclear. This study aims to elucidate the potential mechanisms and key components by which the QRRZF ameliorates asthma.Methods: UPLC-MS/MS was employed to identify the components of the QRRZF; therapeutic targets were predicted with SwissTargetPrediction and Super-Pred. Asthma transcriptomic data were obtained from GEO and differentially expressed genes were identified (GSE43696 and GSE147878). Venn analysis yielded potential targets of the QRRZF against asthma. GO and KEGG enrichment analyses with Metascape identified key pathways; the pathway-associated proteins served as receptors for molecular docking to rank drug-receptor affinities, thereby identifying key anti-asthma components. CETSA was performed on 16HBE cells cultured in vitro to verify docking results. CCK-8 and ELISA assessed the anti-asthma effects of the key components in vitro, and an HDM-induced asthma mouse model evaluated its efficacy in vivo.Results: A total of 21 components and 827 predicted targets were obtained; 1250 asthma-related targets were extracted from GEO, yielding 76 potential therapeutic targets. Enrichment analysis suggested calcium signaling pathway as the main pathway; molecular docking showed Deacetyl Ophiopojaponin A (DOA) bound most tightly to Cav2.3, confirmed by CETSA. In vitro experiments demonstrated that DOA protected 16HBE cells from damage induced by house dust mites, reducing the levels of inflammatory cytokines and alleviating airway inflammation, such IL-6, IL-13, and TNF-α. In vivo experiments showed that DOA reduced inflammatory cell infiltration and airway inflammation in the lung bronchioles, protecting the epithelial barrier and demonstrating significant therapeutic efficacy. These results indicate that DOA exerts potent anti-asthma effects by inhibiting the Cav2.3 calcium channel.Conclusion: This study reveals that calcium signaling pathway is the principal mechanism by which QRRZF improves asthma, and DOA is the key therapeutic component, which alleviates asthma-induced airway inflammation by inhibiting Cav2.3 expression and reducing calcium load in airway epithelial cells. The diagram illustrates two scenarios involving airway epithelium. On the left, a mouse is exposed to HDM, leading to activation of Cav2.3 channels in the airway epithelium. This results in airway inflammation and asthma. In the second scenario, DOA is introduced, which inhibits the activation of Cav2.3 channels, reducing airway inflammation and asthma. The diagram highlights the role of Cav2.3 in airway inflammation and the inhibitory effect of DOA.Diagram showing HDM exposure leading to airway inflammation and asthma, with DOA inhibiting inflammation.Keywords: Qingre Runzao formula, asthma, Deacetyl Ophiopojaponin A, CETSA, calcium signaling pathway
Li et al. (Fri,) studied this question.