Effect of Haoqin Qingdan Tang on influenza A virus through the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway

Shiyun Liang; Jieling Lin; Mengjie Xiao; Tongmei Shi; Yu Song; Tianbo Zhang; Xi Zhou; Runfeng Li; Xin Zhao; Zifeng Yang; Huihui Ti

doi:10.1016/j.phymed.2024.155680

Effect of Haoqin Qingdan Tang on influenza A virus through the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway

Phytomedicine. 2024 Apr 25:129:155680. doi: 10.1016/j.phymed.2024.155680. Online ahead of print.

Authors

Shiyun Liang¹, Jieling Lin¹, Mengjie Xiao², Tongmei Shi¹, Yu Song², Tianbo Zhang³, Xi Zhou², Runfeng Li⁴, Xin Zhao², Zifeng Yang⁵, Huihui Ti⁶

Affiliations

¹ School of Chinese Materia Medica, Guangdong Pharmaceutical University, Guangzhou, 510006, China.
² Guangdong Provincial Key Laboratory of Chemical Measurement and Emergency Test Technology, Guangdong Provincial Engineering Research Center for Ambient Mass Spectrometry, Institute of Analysis, Guangdong Academy of Sciences(China National Analytical Center, Guangzhou, Guangzhou, 510070, China.
³ Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, 94305, USA.
⁴ State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510000, China.
⁵ State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510000, China; Guangzhou Laboratory, Guangzhou, 510000, China; Guangzhou Key Laboratory for Clinical Rapid Diagnosis and Early Warning of Infectious Diseases, Guangzhou, 510000, China; State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Taipa, Macau, 519020, China. Electronic address: jeffyah@163.com.
⁶ School of Chinese Materia Medica, Guangdong Pharmaceutical University, Guangzhou, 510006, China; Guangdong Province Precise Medicine Big Date of Traditional Chinese Medicine EngineeringTechnology Research Center, Guangdong Pharmaceutical University, Guangzhou, 510006, China. Electronic address: tihuihui@126.com.

PMID: 38728923
DOI: 10.1016/j.phymed.2024.155680

Abstract

Objective: Influenza, a viral respiratory illness, leads to seasonal epidemics and occasional pandemics. Given the rising resistance and adverse reactions associated with anti-influenza drugs, Traditional Chinese Medicine (TCM) emerges as a promising approach to counteract the influenza virus. Specifically, Haoqin Qingdan Tang (HQQDT), a TCM formula, has been employed as an adjuvant treatment for influenza in China. However, the active compounds and underlying mechanisms of HQQDT remain unknown.

Aim: The aim of this study was to investigate HQQDT's antiviral and anti-inflammatory activities in both in vivo and in vitro, and further reveal its active ingredients and mechanism.

Methods: In vivo and in vitro experiments were conducted to verify the antiviral and anti-inflammatory activities of HQQDT. Subsequently, the active ingredients and mechanism of HQQDT were explored through combining high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS) analysis and network pharmacology. Finally, the examinations of cell cytokines and signaling pathways aimed to elucidate the predicted mechanisms.

Results: The results indicated that HQQDT exhibited inhibitory effects on influenza viruses A/PR/8/34 (H1N1), A/HK/1/68 (H3N2), and A/California/4/2009 (H1N1) in vitro. Furthermore, HQQDT enhanced the survival rate of influenza-infected mice, reduced the lung index and lung virus titer, and mitigated lung tissue damage in vivo. The proinflammatory cytokine expression levels upon influenza virus infection in PR8-induced A549 cells or mice were suppressed by HQQDT, including IL-6, IL-1β, CCL₂, CCL₄, IP-10, interferon β1 (IFN-β1), the interferon regulatory factor 3 (IRF3), and hemagglutinin (HA). Twenty-two active components of HQQDT against influenza were identified using HPLC-Q-TOF-MS analysis. Based on network pharmacological predictions, the JAK/STAT signaling pathway is considered the most relevant for HQQDT's action against influenza. Finally, western blot assays revealed that HQQDT regulated the protein level of the JAK/STAT signaling pathway in PR8-infected A549 cells and lung tissue.

Conclusion: These findings verified the antiviral and anti-inflammatory effects of HQQDT through JAK-STAT signaling pathway in influenza infections, laying the foundation for its further development.

Keywords: Haoqin Qingdan Tang; Influenza virus; JAK/STAT; Network pharmacology.