Analysis of network pharmacology and experimental verification to explore the mechanism of Modified Guilu Erxian Glue in the treatment of low immune function

Abstract

ObjectiveThe aim of this study was to investigate the therapeutic effect of modified Guilu Erxian Gum (MGEG) on cyclophosphamide (Cy) induced immunosuppression in mice. Based on network pharmacology, the comprehensive mechanism of immune hypofunction was preliminarily discussed. MethodsCyimmunosuppression model was established to observe the effect of MGEG compound on immune injury in mice. The best compound was screened for network pharmacological analysis, and the "drug component" and "disease drug target" networks were established by network pharmacological methods. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were conducted according to the common target between Chinese herbal medicine and low immunity to determine the biological process and signal pathway. The core components and targets obtained by network pharmacology were verified by Western blot analysis (WB). ResultsIn vivo experiments showed that the weight, thymus index, organ index, carbon particle clearance index, phagocytosis index, foot thickening level, macrophage phagocytosis and natural killer cell killing activity of immunosuppressed mice induced by Cy were significantly increased by Codonopsis pilosula, Guilu Erxian Jiaowei Wuxing Powder (CGW) (p<0.05 or p<0.01). In addition, CGW significantly increased the humoral and cellular immunity of immunosuppressed mice (p<0.05 or p<0.01). Based on the network pharmacological analysis, it is predicted that CGW contains 125 potential bioactive components, involving 489 action targets, and 125 common targets that may be related to immunosuppressive diseases. Based on the establishment of protein protein interaction network, five central nodes were identified as active factors involved in major biological functions. David, ClueGO and Metascape enrichment analysis all proved that CGW plays an important role in the regulation of immune function. By KEGG analysis, TNF signaling pathway is considered to be one of the main signaling pathways of CGW in the treatment of immunosuppression, including a wide range of intracellular signaling pathways. NF-κB signaling pathway regulates inflammation and immunity in TNF signaling pathway. WB showed that CGW significantly promoted NF-κB The expression of signal pathway further confirms that Chinese herbal compound can enhance the body's immunity and NF-κB signal path activation. ConclusionBased on network pharmacology and experimental verification, the potential targets and pathways of CGW to improve Cy induced immunosuppression and its potential therapeutic mechanism were revealed, providing a theoretical basis for development of traditional Chinese medicine. Guilu Erxian Gum (龟鹿二仙胶, GEG) is a traditional Chinese medicine prescription, which is widely used to treat low immunity and immune function impairment caused by chemotherapy. However, its potential pharmacological mechanism remains largely unknown.