Siji Kangbingdu Mixture Ameliorates LPS-Induced Acute Lung Injury by Downregulating the TNF/IL-17 Signaling Pathways: A Comprehensive Study Using Transcriptomics, Network Pharmacology and Experimental Verification

Yuan Wang,1 Huiying Zhou,1 Bo Wang,2 Yuxi Liu,1 Kaihua Long,1 Hong Zhang1 1Institute of Chinese Materia Medica, Shaanxi 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 ChinaCorrespondence: Kaihua Long, 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]: Acute lung injury (ALI), as a respiratory condition triggered by various internal and external factors, is characterized clinically by high rates of incidence and mortality. In the current context where respiratory system diseases are on the rise, the incidence of ALI has been increasing annually. Previous research has established the efficacy of traditional Chinese medicine (TCM) in inhibiting ALI. Siji Kangbingdu Mixture (SJM), has demonstrated a clear clinical therapeutic effect in managing respiratory infections. Nevertheless, its mechanism of action in treating ALI is still lacking.Purpose: To clarify the mechanism by which SJM exerts its therapeutic effects in the treatment of ALI.Materials and Methods: In our study, we initially elucidated the chemical constituents of SJM by the UHPLC-Q-Orbitrap HRMS (LC-MS), and established an ALI model. Next, we evaluated how SJM inhibited ALI by measuring the weight curve of mice, the wet-to-dry lung weight ratio (W/D), the HE lung tissue pathological change score, along with the inflammatory factor expression. Then, by employing transcriptomics and network pharmacology approaches, the mechanism of SJM acting on ALI were predicted and analyzed. Finally, the expression and regulatory interactions of the differentially expressed target proteins in the key pathways were verified by qRT-PCR analysis, Western blotting (WB), immunofluorescence and immunohistochemistry.Results: The LC-MS analysis identified 148 active ingredients. In vitro experiments indicated that SJM could significantly reduce the W/D index, pulmonary tissue pathological score, and the inflammatory factor levels. The findings from transcriptomics and network pharmacology suggested that SJM may alleviate ALI as a TNF and IL-17 signaling pathway. Further, through WB, immunofluorescence and immunohistochemistry, we noted significant downregulations in the pulmonary tissue expression of keys proteins in the SJM-H group.Conclusion: SJM has the potential to inhibit the progression of ALI, and its mechanism of action may involve the TNF and IL-17 signaling downregulation. Keywords: Siji kangbingdu mixture, acute lung injury, network pharmacology, transcriptomics, TNF signaling pathway, IL-17 signaling pathway