Mechanism of Action of San Ren Decoction in PLWH Treatment Determined Using Network Pharmacology Combined with...

To explore the mechanism of action of San Ren Decoction (SRD) in people living with HIV (PLWH) treatment using network pharmacology, molecular docking technology, and cellular experiments. The active ingredients and potential targets of the Chinese traditional herb in SRD were determined from traditional Chinese Medicine Systems Pharmacology and Bioinformatics Analysis Tool for Molecular Mechanism of traditional Chinese medicine. The therapeutic targets of SRD in AIDS were identified using GeneCards, OMIM, DisGeNET, and DrugBank. The overlap between disease and component targets was determined to identify the potential targets of SRD in AIDS. The network of "Chinese herbs-active ingredients-targets" for SRD was accessed using Cytoscape. A protein-protein interaction network was prepared using STRING. The Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were used to perform enrichment analysis of signaling pathways. Molecular docking experiments and visualization of results were performed using Auto Dock Vina and PyMOL. Based on the results of network pharmacology, a drug-containing serum was prepared through cellular experiments. Peripheral blood mononuclear cells (PBMCs) were isolated from blood samples of PLWH and divided into three groups: the PLWH group, the PLWH + PI3K/Akt inhibitor group, and the PLWH + SRD drug-containing serum group (represented by PLWH, ZSTK474, and SRD, respectively). Healthy human PBMCs were used in the control group. After grouped culturing, quantitative polymerase chain reaction and enzyme-linked immunosorbent assay were performed to detect and confirm gene and protein expression in each group. Quercetin, luteolin, myristic acid, honokiol, arachidonic acid, and other core components were the active ingredients in SRD. The core targets of SRD in AIDS included CAV1, SRC, HSP90AA1, AKT1, PI3K, STAT1, and RAF1. Gene ontology functional enrichment analysis revealed the positive regulation of gene expression, the response to foreign stimuli, and other observations. KEGG pathway enrichment analysis showed the involvement of the PI3K/Akt, TLR, and other pathways. Molecular docking results indicated that the primary active ingredients of SRD exhibited stable binding with the core proteins. In vitro and in vivo experiments showed that the mRNA and protein levels of AKT1, Caspase8, mTOR, PI3K, STAT1, and Bcl-2 were higher in PLWH. SRD may help regulate PLWH by inhibiting the PI3K/Akt signaling pathway. The results of this study indicated that SRD may play a role in PLWH treatment through multiple components and multiple targets to regulate the PI3K/Akt signaling pathway.