Xi Tian,1,2,* Qingfei Cui,3,* Jinhuan Wei,1 Qian Zhang,1 Huiyi Zhang,1 Mengxin Yang,1 Yiqi Xing,4 Yukun Niu,1 Wenyu Li,1 Nan Wang,1 Yiran Jin,5 Yingfeng Du1 

1Department of Pharmaceutical Analysis, School of Pharmacy, Hebei Medical University, Shijiazhuang, Hebei, People’s Republic of China; 2Institute of Cash Crop, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang, Hebei, People’s Republic of China; 3Shijiazhuang Biomedical Academician Workstation, Shijiazhuang Yiling Pharmaceutical Co., Ltd., Shijiazhuang, Hebei, People’s Republic of China; 4Architectural Engineering Institute, Hebei Vocational University of Industry and Technology, Shijiazhuang, Hebei, People’s Republic of China; 5Department of Pharmacy, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, People’s Republic of China

*These authors contributed equally to this work

Correspondence: Yingfeng Du, Department of Pharmaceutical Analysis, School of Pharmacy, Hebei Medical University, Shijiazhuang, Hebei, People’s Republic of China, Tel +86-311-86265625, Fax +86-311-86266419, Email 17700941@hebmu.edu.cn Yiran Jin, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, People’s Republic of China, Email jinyiran@sohu.com

Background: In traditional Chinese Medicine (TCM), Zhimu (Anemarrhena asphodeloides Bunge, ZM) - Chuanbeimu (Fritillaria Cirrhosae Bulbus, CBM) (ZC) is one of the most classical herb pairs used in the treatment of lung diseases such as asthma. This study aimed to investigate how ZC affects asthma and its mechanism.
Methods: Asthma model rats were sensitized by ovalbumin. The anti-asthma efficacy of ZM, CBM and ZC were evaluated through analysis of lung function, pathological sections and biochemical indices. Metabolomics based on UHPLC-QTOF-MS was conducted to determine the synergistic anti-asthma effect of combination therapy. Asthma targets and mechanism prediction were performed using network pharmacology. Then, the potential anti-asthma mechanism of ZC was explored using RT-qPCR.
Results: According to the lung function test, Hematoxylin-Eosin Staining experiment, ZC herb pair had an obvious anti-asthma effect over either ZM or CBM alone. It has also been demonstrated that positive effect of ZC against Th1/Th2 immune imbalance. Both metabolomics and network pharmacology were highly enriched in the arachidonic acid metabolism pathway. The mRNA expression levels of ALOX5, PLA2G4A and CYP1A2, critical targets in arachidonic acid metabolism, were significantly down-regulated by RT-qPCR.
Conclusion: By reducing the expression of cytokines and chemokines mediated by the arachidonic acid metabolism pathway, ZC could alleviate OVA-induced asthma in vivo. It was the first to demonstrate the complex mechanism of ZC for the treatment of asthma. Meanwhile, a new paradigm was established for evaluating the pharmacological effects of TCM drugs for asthma based on multiple mechanisms.

Keywords: Anemarrhena asphodeloides Bunge-Fritillaria Cirrhosae Bulbus, anti-asthma, combination mechanisms, network pharmacology, metabolomics, RT-qPCR