Effect and Mechanism of Nebulized Inhalation of Reduning Injection on Improving Pulmonary Inflammation in Mice with Smoke-induced Chronic Obstructive Pulmonary Disease
Objective:To investigate the therapeutic effect and mechanism of Reduning Injection(RDN)on the pulmonary inflammation in a cigarette smoke-induced chronic obstructive pulmonary disease(COPD)mouse model.Methods:The contents of chlorogenic acid and geniposide in RDN were determined using high-performance liquid chromatography(HPLC).A cigarette smoke-induced COPD mouse model was established and treated with nebulized inhalation of RDN.Wright's staining was used to detect the types and numbers of inflammatory cells in bronchoalveolar lavage fluid(BALF).The pathological changes in lung tissues were observed using hematoxylin-eosin(HE)staining.The expression of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and IL-6 in lung tissues was detected using immunohistochemical(IHC)staining.Transcriptomic analysis was used to screen differentially expressed genes in the lung tissues of the control group,model group,and RDN high-dose group,followed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.The expression levels of related proteins in the Toll-like receptor 4(TLR4)signaling pathway were determined by Western blot.Results:The concentrations of chlorogenic acid and geniposide in RDN were 4.75 and 7.60 mg·mL-1,respectively.RDN reduced the number of macrophages and neutrophils in BALF and improved pathological changes such as interstitial and alveolar hemorrhage,alveolar wall edema,and inflammatory infiltration in the lung tissues of mice.RDN inhibited the expression of inflammatory factors TNF-α,IL-1β,and IL-6 in lung tissues.Transcriptome sequencing results showed that there were 149 common differentially expressed genes among the three groups.KEGG enrichment analysis indicated that these genes were mainly involved in the IL-17 signaling pathway,Toll-like receptor signaling pathway,TNF-α signaling pathway,and nuclear factor-κB(NF-κB)signaling pathway.RDN reduced the expression of TLR4 in lung tissues and inhibited the phosphorylation levels of downstream related proteins p38,JNK,extracellular signal-regulated kinase(ERK),and p65.Conclusion:RDN can improve the inflammatory response in the lung tissues of cigarette smoke-induced COPD model mice,and its mechanism is related to the inhibition of the TLR4 signaling pathway.
万方数据
维普
