Biao Lei,1– 3,* Wenbo Huang,4,* Wei Peng,1,* Jingwen Mu,2,3,* Yi Liu,2,3,* Chunxian Zhou,2,3 Ting Cheng,2,3 Xiaojiao Wei,2,3 Lingyun Pan,5 Yuejuan Zheng,2,3 Bangjiang Fang1 

1Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, People’s Republic of China; 2The Research Center for Traditional Chinese Medicine, Shanghai Institute of Infectious Diseases and Biosecurity, School of Integrative Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, People’s Republic of China; 3Center for Traditional Chinese Medicine and Immunology Research, School of Integrative Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, People’s Republic of China; 4State 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, Guangdong, People’s Republic of China; 5Experiment Center for Science and Technology, Shanghai University of Traditional Chinese Medicine, Shanghai, People’s Republic of China

*These authors contributed equally to this work

Correspondence: Yuejuan Zheng, The Research Center for Traditional Chinese Medicine, Shanghai Institute of Infectious Diseases and Biosecurity, School of Integrative Medicine, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Road, Pudong New District, Shanghai, 201203, People’s Republic of China, Email 13641776412@163.com Bangjiang Fang, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 725 Wanping South Road, Xuhui District, Shanghai, 200000, People’s Republic of China, Email fangbji@163.com

Purpose: Shu-Feng-Jie-Biao formula (SFJBF) has been used to treat acute respiratory infections for a dozen years. This study aimed to explore its mechanisms and effects for the treatment of influenza.
Methods: Network pharmacology was used to explore the underlying mechanism of SFJBF against influenza. The protective effects of SFJBF in vivo were evaluated by lung indexes, body weight loss and pathological changes in lungs. The anti-inflammatory effects in vivo were evaluated by flow cytometry and ELISA. RAW264.7 cells stimulated with imiquimod (R837) were used to determine the anti-inflammatory effects of SFJBF. Neutrophils isolated from bone marrow were activated by phorbol 12-myristate 13-acetate (PMA) to validate the effects of the active components of SFJBF.
Results: SFJBF protected body weight loss, decreased lung indexes, reduced total protein content in lungs and mitigated pathological changes in mice. SFJBF inhibited the expression of chemokines (Cxcl2 and Ccl2) and cytokines (Il1b and IL-6) accompanied by the decreased infiltration of neutrophils in lungs. SFJBF inhibited the expression of iNOS and MPO in lungs. The synergistic role of OSV and SFJBF was exhibited by suppressing virus-induced cytokine expression and reducing the infiltration of inflammatory monocytes in lungs. SFJBF inhibited the phosphorylation of ERK1/2 and NF-κBp65, thereby reducing the secretion of MIP-2, TNF-α, MCP-1 and CCL5 in vitro The active components of SFJBF, including baicalin and wogonin, reduced the production of reactive oxygen species (ROS), MIP-2, MCP-1, and IL-6 in vitro.
Conclusion: SFJBF ameliorated virus-induced lung injury by suppressing overactivated immune responses via NF-κB and ERK MAPK signaling pathways, thereby protecting mice from influenza virus infection. SFJBF could be considered a potent therapeutic agent for treating influenza.

Keywords: traditional Chinese medicine, TCM, anti-inflammatory, influenza virus, acute lung injury, NF-κB signaling pathway, ERK MAPK signaling pathway