Study On The Mechanism Of Xingbei Zhike Granules In The Treatment Of Acute Lung Injury

Object:Based on network pharmacology,to explore the target and mechanism of Xingbei Zhike Granules in the treatment of acute lung injury,evaluate its efficacy by animal experiments,and verify the key target of network pharmacology,to provide new ideas for the treatment of acute lung injury with traditional chinese medicine.Methods:1.The TCMSP database was used to screen the drug composition and targets of Xingbei Zhike Granules.Gene Cards,OMIM database were screened with the keyword “Acute lung injury”to obtain disease targets for acute lung injury.Venn diagrams are created to obtain intersectional targets for drug diseases.The STRING database was used to construct proteinprotein interaction(PPI)networks by feeding in relevant intersecting targets.Cytoscape3.9.1 was used to construct network maps.The open source bioinformatics software Bioconductor was used for GO enrichment and KEGG pathway analysis.2.C57BL/6J male mice were divided into blank group,model group and Xingbei Zhike Granules groups(3g/kg/d and 6 g/kg/d).The mice in Xingbei Zhike Granule groups were gavaged with Xingbei Zhike Granules and the mice in the blank group and the model group were given an equal volume of distilled water daily for 1 week.Except for the blank group,mice were injected intraperitoneally with LPS(10 mg/kg)to construct a model of acute lung injury in mice.Then the modeling group was divided into the model group,the low-dose group(3 g/kg/d)and the high-dose group(6 g/kg/d)according to the random number table method.The mice were executed at 6 h,12 h and 24 h after modeling,and the lung tissues of each group were stained with HE to observe the pathological development of lung tissue.Real-time quantitative PCR was used to verify the expression of IL-6,IL-1β and TNF-α in lung tissue.The mice were executed at 6 h,and the wet/dry weight ratio and superoxide dismutase(SOD)activity of lung tissues were measured.ELISA was used to detect the levels of proinflammatory cytokines IL-6,IL-1β and TNF-α in plasma and alveolar lavage fluid(BALF).Real-time quantitative PCR was used to verify the expression of NLRP3,caspase-1,GSDMD and IL-18 regulatory molecules in lung tissue.Results:1.A total of 254 ingredients of Xingbei Zhike Granules were searched from TCMSP database.After the screening of ADME parameters,a total of 183 active components were found to be in line with oral bioavailability(OB)and drug(DL)after removing duplicates.2723 drug targets were retrieved,and 267 drug targets were obtained after standardization and removing duplicates by UNIPROT.A total of 9838 related targets were obtained by searching the disease target database.According to the correlation criteria,a total of 9586 hypertension related targets were obtained by UNIPROT standardization and EXCEL software after removing duplicates.The PPI network was constructed with 68 key targets including JUN,MAPK3,NLRP3,STAT3,AKT1,caspase1,GSDMD,FOS,TNF and RXRA.GO analysis showed 4673 biological processes related to response to drug、cellular response to chemical stress、reactive oxygen species metabolic process、response to oxidative stress、response to lipopolysaccharide and so on.KEGG analysis showed 283 pathways related to AGE-RAGE signaling pathway in diabetic complications、Lipid and atherosclerosis、c GAS-STING signaling pathway and so on.2.Compared with the blank group,in the model group,the lung tissue damage was progressively aggravated,the expression levels of IL-6,TNF-α and IL-1β in the lung tissues of mice were significantly higher(P<0.05),the lung wet/dry weight ratio was significantly higher(P<0.05),the activity of SOD in lung tissues was slightly reduced,the levels of pro-inflammatory cytokines IL-6,IL-1β and TNF-α in plasma and BALF were significantly higher(P<0.05),and the expression levels of NLRP3,caspase-1,GSDMD and IL-18 m RNA,the regulatory molecules related to the carcinogenesis pathway,were higher in lung tissues;compared with the model group,in Xingbei Zhike Granule groups,the lung tissue damage was progressively reduced,the expression levels of IL-6,TNF-α and IL-1β in the lung tissues were significantly lower(P<0.05),the wet/dry weight ratio of lungs was significantly lower(P<0.05),the activity of SOD in lung tissues was increased,the levels of pro-inflammatory cytokines IL-6,IL-1β and TNF-α in plasma and BALF were significantly lower(P<0.05),The expression levels of NLRP3,caspase-1,GSDMD and IL-18 m RNAs in lung tissues were reduced.Conclusion:1.Network pharmacology analysis: Xingbei Zhike Granules may treat acute lung injury by modulating inflammatory responses and related signalling pathways such as immune regulation.2.Animal experiments: Xingbei Zhike Granules had some ameliorating effect on LPS-induced acute lung injury in mice,and the mechanism may be related to the inhibition of cell scorching.