Network Pharmacology, Molecular Docking, and Experimental Validation of the Mechanism of Jiedu Xiaoliu Formula against Diffuse Large B-Cell Lymphoma.

Abstract

Diffuse Large B-Cell Lymphoma (DLBCL) is the most common Bcell lymphoma type. Detoxification and tumor elimination formula, a herbal compound, can potentially treat lymphoma. In this study, network pharmacology and molecular docking approaches were utilized to reveal the potential mechanism of the Jiedu Xiaoliu formula (JDXLF) against DLBCL. Active compounds and targets of JDXLF were obtained from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. Targets related to DLBCL were retrieved from GeneCards and Online Mendelian Inheritance in Man (OMIM) databases. Protein- Protein Interaction (PPI) networks were established to screen core targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using R 4.2.2. Model interactions between potential disease targets and pharmacologically active compounds were determined by molecular docking. Screening of 14 herbal active ingredients yielded 129 active compounds and 1414 disease targets for DLBCL. GO annotations showed that the effects of JDXLF were related to protein phosphorylation and reactive oxygen species response. KEGG pathway enrichment analysis indicated that the detoxification and elimination of tumors formula mainly regulated apoptosis pathways. Nobiletin showed good interaction with AKT1, TP53, and CASP3, and the cell counting kit-8 (CCK-8) assay confirmed that nobiletin inhibited the proliferation of SU-DHL-4 cells. Western blot analysis showed that nobiletin downregulated the expressions of p-PI3K, p- AKT, and BCL-2 proteins and upregulated those of cleaved-caspase3 and BAX. Our findings preliminarily suggested that the active ingredient of JDXLF, nobiletin, may induce apoptosis in Diffuse Large B-Cell Lymphoma SU-DHL-4 cells by regulating the PI3K/AKT signaling pathway.