Mechanism Of Quyang Huashi Decoction In Treating Perianal Eczema And Pruritus Based On Network Pharmacology Approach

Objective:Based on the network pharmacological approach,the mechanism of Quyang Huashi Decoction(QYHSD)in treating perianal eczema and pruritus was explored,so as to find the potential targets and signal pathways of QYHSD.Further experiments in vitro were carried out to provide more evidences for the scientific effectiveness of this approach and more foundations for the research and development of new drugs of traditional Chinese medicine.Methods:Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine(BATMAN-TCM)were used to predict the chemical components of QYHSD,and the corresponding gene targets were searched by UniProt,GeneCards and other databases.The target network map of QYHSD-active compounds-diseases was constructed by the visual network pharmacology tool Cytoscape and the protein-protein interaction network(PPI)of QYHSD was constructed by STRING software.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)of the compound target network were performed by DAVID platform.To reveal the potential targets and signal pathways of QYHSD in the treatment of perianal eczema and pruritus.The anti-inflammatory mechanism of QYHSD was studied by human keratinocyte(HaCaT).CCK8 activity assay was used to evaluate the 50%Inhibiting Concentration(IC50)of QYHSD in HaCaT cells.Inflammatory response was induced by stimulation of tumor necrosis factor-?(TNF-?)and interferon-y(IFN-y).The expression of IL-6,IL-1?,TNF-? inflammatory chemokines in induced HaCaT cells was measured by enzyme-linked immunosorbent assay(ELISA).Western blot was used to detect the phosphorylation levels of signal transducer and activator of transcription 3(STAT3)and Janus kinase 2(JAK2)Results:It showed that there were 83 potential targets of QYHSD,such as PIK3CA,STAT3,EGFR,IL-6,IL-10,JAK2,JAK1,TNF and so on.At the same time,functional annotation analysis of target protein gene showed biological processes molecular functions and cellular component.KEGG signal pathway enrichment analysis shows that most genes are enriched in PI3K-Akt signaling pathway,cytokine-cytokine receptor interaction,inflammatory bowel disease,Jak-STAT signaling pathway,TNF signaling pathway,Toll-like receptor signaling pathway,FoxO signaling pathway,HIF-1 signaling pathway,MAPK signaling pathway and so on.In cell experiment,QYHSD inhibited the proliferation of HaC aT cells in a concentration-dependent manner,and IC50 was 672.6?g/ml.In the cells stimulated by TNF-? and IFN-?,it was observed that QYHSD at high,middle and low concentrations could inhibit the production and expression of IL-6,IL-1?,TNF-? inflammatory factors,as well as the expression and phosphorylation of STAT3 and JAK2.Conclusions:The therapeutic effect of QYHSD on perianal eczema and pruritus may be multi-component,multi-target and multi-pathway.It is speculated that mainly through a number of immunological pathways,receptor signal pathways,cytokine/chemokine pathways,etc.,to participate in biological processes such as immunity,inflammation,apoptosis,gene transcription,gene expression and so on.Furthermore.the results of in vitro experiments showed that QYHSD exerted its anti-inflammatory effect by inhibiting the phosphorylation of JAK2 and STAT3 and blocking the JAK2/STAT3 signal pathway.