Ligusticum chuanxiong, Salvia miltiorrhiza and Morindae officinalis jointly enhanced the antidepressant effect of Kaixin San

Background Depression, a prevalent global mental health disorder, is characterized by persistent low mood, anhedonia, and high relapse rates. Current mainstream treatments often have limitations. In contrast, Traditional Chinese Medicine (TCM) offers potential advantages through multi-target effects and favorable safety profiles, presenting a promising approach for developing novel antidepressants. Objective This study employed network pharmacology, with in vivo and in vitro validation, to compare the active components and target sites of Kaixin San (KXS) and its modified formulation Jia-Wei Kaixin San (JWKXS), as well as to compare their mechanisms and characteristic advantages of antidepressant effects. Methods High-performance liquid chromatography (HPLC) chromatographys of KXS and JWKXS were established based on 3,6′-disinapoyl sucrose, α-asarone, ginsenoside Rb1, dehydrotumulosic acid, monotropein, ferulic acid, and salvianolic acid B. Active components were screened from the TCMSP database, potential antidepressant targets were analysed via the DisGeNet database, and a herb-compound-target network was constructed. Protein-protein interaction networks were built using STRING and analyzed topologically, followed by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Oxidative stress-induced SH-SY5Y cells model in vitro and acute-stress mice models in vivo were established to validate the similarities and differences between KXS and JWKXS. Results For KXS, 200 active components and 358 targets were identified, including 72 potential antidepressant targets. JWKXS contained 436 active components and 449 targets, with 84 potential antidepressant targets. KEGG pathway analysis showed that KXS mainly acts on pathways related to neuroactive ligand-receptor interactions, cAMP signaling, and some pathways associated with cardiovascular and lipid metabolism. In addition to sharing common pathways with KXS. JWKXS demonstrated more diversified signaling pathways, particularly those related to vascular endothelial dysfunction, such as the Rap1, Ras, and EGFR signaling pathways. In an oxidative stress-induced SH-SY5Y cells model, JWKXS significantly enhanced neuronal cell viability ( P < 0.001), exhibiting dose-dependent neuroprotective effects superior to KXS. In acute stress models (tail suspension and forced swimming tests in mice), both formulations significantly reduced immobility time as compared to that of control group (2 h–72 h, P < 0.001; 7 d, P < 0.01) with no significant differences between them. Both formulations upregulated serotonin (5-HT) and downregulated hypoxia-inducible factor (HIF-1α) levels, with JWKXS showing greater efficacy. Additionally, high-dose JWKXS significantly downregulated MAPK3 ( P < 0.05) and ESR1 protein levels ( P < 0.01) and revised brain-derived neurotrophic factor (BDNF) ( P < 0.001) and vascular endothelial growth factor (VEGF) levels ( P < 0.01). Conclusion Compared with the basic KXS formulation, JWKXS, by further addition of Ligusticum chuanxiong, Salvia miltiorrhiza , and Morindae officinalis , do not only enhance the original antidepressant effects but also more effectively regulates potential targets and pathways related to vascular endothelial function. These findings suggest that JWKXS offer advantages in treating depression comorbid with cardiovascular and cerebrovascular diseases.