Pharmacodynamics and molecular mechanism of Jinzhen Oral Liquid in treating bronchial asthma
Objective To establish a mouse bronchial asthma model and a human non-small cell lung cancer cell(A549)inflammation model,and to study the pharmacological efficacy and molecular mechanism of Jinzhen Oral Liquid(JZOL)on bronchial asthma.Methods Sixty-six female BALB/c mice were randomly divided into control group and model group.After the asthma model was established by the ovalbumin(OVA)sensitization method,mice in the model group were divided into model group,JZOL high,medium,and low dose group,and dexamethasone group(Dex).Whole-body volumetric tracing system was used to determine the penh value of airway response in mice.HE staining was used to observe the histopathology of lungs in mice.The number of inflammatory cells in alveolar lavage fluid(BALF)was detected by Riesling-Jimsa staining.ELISA was performed to detect tumor necrosis factor-α(TNF-α)in BALF and immunoglobulin E(IgE)and immunoglobulin G1(IgG1)concentrations in serum.A cellular inflammation model was established by using 20 ng·mL-1 interleukin(IL)-1β induced A549 cells for 24 h.The cells were randomly divided into control group,model group,and four mass concentration JZOL-treated groups(5.39,2.69,1.35,and 0.67 mg mL-1).The MTS assay was used to determine the pharmacotoxicity of JZOL on A549 cells.RNA-seq technology was used to perform transcriptomics analysis of six experimental groups to screen differential genes,which were subjected to GSEA pathway enrichment analysis,KEGG pathway enrichment analysis and GO pathway enrichment analysis,and the key genes screened were experimentally verified by qRT-PCR experiments.Results In vivo experiments showed that JZOL was able to improve inflammatory cell infiltration in mouse lung tissues,reduce the Penh value of airway hyperresponsiveness in mice,decrease the number of macrophages,eosinophils and neutrophils in BALF,and decrease the concentrations of TNF-α in BALF and IgE and IgG1 in serum.In vitro experiments showed that the maximum nontoxic concentration of JZOL on A549 cells was 5.39 mg·mL-1.Gene enrichment analysis showed that a total of 46 inflammation model-related pathways were obtained in the model group compared with the control group,and the percentage of the reversal of these pathways in the groups treated with different concentrations of JZOL was 34.8％(5.39 mg·mL-1),50％(2.69 mg·mL-1),32.6％(1.35 mg mL-1)and 26％(0.67 mg·mL-1),respectively.KEGG enrichment analysis of differentially expressed genes(DEGs)showed that a total of 58 pathways were enriched in model group compared with control group,and a total of 32 pathways were enriched in JZOL-treated group compared with the model group,reversing IL-17,TNF,and nucleotide oligomerization structural domain(NOD)-like receptor signaling pathways.The qRT-PCR validation of chemokine ligand 1(CXCL1),chemokine ligand 2(CXCL2),and lipid carrier protein 2(LCN2)in the IL-17 signaling pathway was consistent with the transcriptome sequencing results.Conclusion JZOL has significant therapeutic efficacy in mouse model of bronchial asthma induced by OVA,effectively alleviating airway hyperresponsiveness and suppressing inflammatory responses in lung tissues in asthmatic mice.The anti-inflammatory effect was likely achieved by regulating the expression of several key genes,including CXCL1,CXCL2 and LCN2,which in turn affected the IL-17 signaling pathway.
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