Qingwen Hufei Granules Attenuate Lipopolysaccharide-Induced Acute Lung Injury by Suppressing NLRP3 Inflammasome-Dependent Pyroptosis

Abstract

Kaihua Long,1,* Yun Yang,2,* Yuan Wang,1 Bo Wang,2 Huiying Zhou,1 Yuxi Liu,1 Ye Li,1 Shuanzhu Yang,1 Liping Cao,1 Tingting Huang,2 Yang Liu,1,3,4 Hong Zhang1 1Shaanxi Academy of Traditional Chinese Medicine (Shaanxi Provincial Hospital of Traditional Chinese Medicine), Xi’an, People’s Republic of China; 2College of Life Sciences, Northwest University, Xi’an, People’s Republic of China; 3Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, Beijing, People’s Republic of China; 4Shaanxi Panlong Pharmaceutical Group Limited by Share Ltd., Xi’an, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yang Liu, Shaanxi Academy of Traditional Chinese Medicine (Shaanxi Provincial Hospital of Traditional Chinese Medicine), No. 4 Xihuamen, Lianhu District, Xi’an, 710003, People’s Republic of China, Email [email protected] Hong Zhang, Shaanxi Academy of Traditional Chinese Medicine (Shaanxi Provincial Hospital of Traditional Chinese Medicine), No. 4 Xihuamen, Lianhu District, Xi’an, 710003, People’s Republic of China, Email [email protected]: Currently, clinical treatments for ALI include vasodilators, glucocorticoids, and mechanical ventilation; however, these therapies are associated with various adverse reactions. Therefore, there is a need to develop safer and more effective treatment options. Traditional Chinese Medicine (TCM), particularly Qingwen Hufei Granules (QHG), has demonstrated efficacy in treating respiratory disorders, including upper respiratory tract infections and influenza. However, the active components and mechanism of action of QHG remain unclear.Methods: Lipopolysaccharide (LPS) induced ALI mouse model. We analyzed the main pharmacodynamic components of QHG by HPLC, determined the effects of QHG on the cell viability and inflammatory factors of RAW264.7 cells, and recorded the body weight and lung tissue wet/dry (W/D) of ALI mice. Inflammatory factors in bronchoalveolar lavage fluid (BALF), serum, and lung tissue were determined using ELISA, and the protective effect of hematoxylin and eosin (H&E) staining of lung tissue was studied. Transcriptomic, qRT-PCR, Western blotting, immunohistochemistry, and immunofluorescence analyses were used to analyze QHG and explore the mechanism of ALI damage reduction.Results: The findings demonstrated that QHG effectively mitigated inflammatory cell invasion and pulmonary edema. Moreover, it diminished the concentrations of IL-6, TNF-α, and IL-1β in lung tissue, serum, and BALF. Furthermore, QHG notably decreased the levels of NO, reactive oxygen species(ROS), IL-6, IL-1β, and TNF-α in the supernatants of RAW264.7 cells. Transcriptomic analysis of the lung tissue revealed that QHG primarily enriched the NOD-like receptor (NLR) signaling pathway. qRT-PCR, Western blotting, immunohistochemistry, and immunofluorescence experiments confirmed that QHG mitigated ALI damage through the NLR signaling pathway. Furthermore, seven key components in QHG were determined by using HPLC.Conclusion: Our study demonstrated that QHG effectively mitigated LPS-induced ALI, and provided preliminary insights into its mechanism of action and material basis. Keywords: acute lung injury, lipopolysaccharide, Qingwen Hufei granules, transcriptomics, NOD-like receptor-signaling pathway

Keywords

• acute lung injury
• lipopolysaccharide
• Qingwen Hufei Granules
• transcriptomics
• NOD-like receptor-signaling pathway