Gualou-Xiebai-Baijiu Decoction Ameliorates Atherosclerosis by Inhibiting the ARG1/ <scp>l</scp> -Arg/NO Pathway: A Study Based on Network Pharmacology

This study explored the potential mechanisms of action of Gualou-Xiebai-Baijiu Decoction (GXBD) in the treatment of atherosclerosis (AS) by integrating computational analyses with preliminary animal experiments. The putative targets of blood-absorbed components in GXBD were obtained and then intersected with AS-related targets, followed by protein-protein interaction network construction, core target identification, and GO and KEGG enrichment analyses. Targets presenting potential causal associations with AS were determined with Mendelian randomization (MR) analyses. Binding stability between candidate compounds and key targets was evaluated with molecular docking and molecular dynamics (MD) simulations. Finally, a mouse model of AS was established for in vivo validation. A total of 379 targets of six blood-absorbed components in GXBD and 1975 AS-related targets were identified, among which 154 were overlapping genes and 64 were further defined as core targets. Enrichment analysis results indicated the involvement of pathways including fluid shear stress, PI3K-Akt, and focal adhesion. Among the targets of GXBD, <i>ARG1</i>, <i>CCR3</i>, <i>EPHB4</i>, <i>MAPK9</i>, <i>MCL1</i>, and <i>PDK1</i> showed significant causal associations with AS, with <i>ARG1</i> determined as the key target. Molecular docking and MD simulations demonstrated that Chrysoeriol glucuronide might stably interact with <i>ARG1</i>. In ApoE^-/- mice, GXBD treatment significantly decreased <i>ARG1</i> expression in the aorta, increased serum l-Arg and NO levels, improved lipid profiles, and reduced aortic plaque burden. These findings provide preliminary evidence that GXBD may ameliorate AS-related pathological changes via the <i>ARG1</i>/l-Arg/NO pathway, offering mechanistic insights into the multicomponent actions of GXBD in AS treatment and highlighting further validation directions.