基于单细胞测序技术探讨柴胡清肝汤干预三阴性乳腺癌免疫微环境潜在分子机制OACSTPCD

Objective:To analyze the potential molecular mechanism of Chaihu Qinggan Decoction in the intervention of triple negative breast cancer(TNBC)immune microenvironment by using single cell RNA sequencing related bioinformatics technology.Methods:sin-gle-cell RNA sequencing data related to TNBC was collected and principal component analysis and t-SNE clustering were performed by using the Seurat program in R language.The active ingredients of Chaihu Qinggan Decoction through multiple databases and litera-ture were obtained,and the intersection targets of specific genes that interfere with various cell subsets in TNBC were extracted.The Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes enrichment analysis(KEGG)were performed.Use the immune cell matrix of TNBC in TCGA database for immune infiltration cell level analysis.Finally,validate the molecular docking of key active ingredients of Chaihu Qinggan Decoction with immune related target proteins.Results:A total of 14 cell clusters were obtained.The ac-tive ingredients in Chaihu Qinggan Decoction,such as quercetin,burdock phenol,burdock glycoside and kaempferol,may act on targets such as CD14,IL-1β,SYK and ITGA to intervene in M2 macrophages,involving nuclear factor-kappa B signaling pathway(NF-κB),phosphatidylinositol 3-kinase-protein kinase B(PI3K-Akt),Toll like receptor signaling and other pathways;The active ingre-dients above may act on targets such as CXCR3,JAK,STAT and ITGA to intervene in CD4+T cells,involving janus kinase-signal transducer and activator of transcription(JAK-STAT)signaling pathway,chemokine signaling pathway,PD-L1 and other related pathways.In addition,most target genes are correlated with the corresponding immune infiltrating cell level in the immune matrix in TC-GA.Quercetin,chrysin and kaempferol mainly intervene in other cell subsets in the TNBC microenvironment by regulating MAPK,Wnt,tumor necrosis factor,cAMP,AGE-ARGE and PI3K-Akt signaling pathways through proteins such as JUN,CALM1,JAK1,IL-6 and INSR.In the molecular docking results,more than 50%of the active ingredients have strong docking activity with the target protein.Conclusion:The molecular mechanism of key active ingredients in Chaihu Qinggan Decoction intervening in the immune microenviron-ment of TNBC involves multiple targets and cellular components,as well as multiple types of pathways such as tumor,immune and in-flammatory response.