Identification of Key Targets of Herbal Compounds for Liver Fibrosis Using Network Pharmacology Combined With...

Chinese herbal compound prescriptions have demonstrated efficacy in preventing and treating liver fibrosis (LF), though their mechanisms remained unclear. This study is aimed at identifying diagnostic biomarkers and elucidating the molecular mechanism underlying the effects of the TCM prescription on LF. LF-related datasets (GSE162694, GSE84044, and GSE136103) were obtained from a public database. Active ingredient-related target genes (AIRTGs) and LF-related target genes (LFRTGs) were intersected with differentially expressed genes (DEGs) between the LF and normal control (NC) group to select candidate genes. Subsequently, biomarkers for LF diagnosis were determined using Boruta and LASSO algorithms, receiver operating characteristic (ROC), and expression analyses. A nomogram was constructed to evaluate the capability of these biomarkers for predicting LF risk. Furthermore, GSEA and immunoinfiltration analysis were conducted, along with single-cell analysis to identify relevant cell types in LF. COL3A1 and ALOX5 were identified as diagnostic biomarkers for LF, and the nomogram was proven effective in predicting LF risk. GSEA showed that COL3A1 might play vital roles in cell growth and differentiation, extracellular matrix organization, and cell-matrix interactions. The functions of ALOX5 might be associated with cell-cell interaction, cytoskeletal regulation, and so forth. Immunoinfiltration analysis revealed that activated dendritic cells (DCs) were highly infiltrated, whereas monocytes were less infiltrated in LF. COL3A1 expression was positively correlated with monocytes, but both COL3A1 and ALOX5 showed negative correlations with activated DCs. Single-cell analysis identified nine cell types, with macrophages, B cells, and mesenchyme cells emerging as key cell types. Cell communication analysis demonstrated stronger interactions between macrophages and mesenchymal cells in the LF group. Pseudotime analysis unveiled that the expression of ALOX5 was upregulated and then downregulated, whereas that of COL3A1 was gradually downregulated during the midstage and stabilized thereafter. COL3A1 and ALOX5 may serve as biomarkers for the diagnosis and treatment of LF with Chinese herbal compound prescriptions, contributing to more accurate diagnosis and improved LF therapy.