Guanxin Ⅱ alleviates atherosclerotic comorbid depression via modulating AKT1-STAT3-TLR4 axis and ATG3-mediated...

Guanxin Ⅱ (GXⅡ) is a traditional Chinese herbal formula, which has the functions of activating blood to resolve stasis and move qi to relieve pain. Qi and blood stagnant is regarded as the main pathogenesis of atherosclerosis (AS) and depression, but the specific mechanism of GXⅡ in the treatment of AS comorbid depression (co-depression) remains unclear.

This study aimed to investigate the therapeutic mechanisms of GXⅡ in ameliorating AS co-depression through multi-target network regulation and ATG3-mediated autophagy pathway, integrating pharmacochemical profiling, network pharmacology, and experimental validation.

The absorbed components of GXⅡ in mouse plasma were identified by UPLC-Q-TOF/MS. ApoE^-/- mice were fed with a high fat diet (HFD) combined with restraint stress for 16 weeks, and GXⅡ was administered orally. Anti-AS effects were evaluated by serum lipid panel, aortic Oil Red O staining and HE staining. Antidepressant effects were assessed by open field test (OFT), sucrose preference test (SPT), and tail suspension test (TST). Network pharmacology was employed to predict core targets, which were validated by molecular docking and molecular dynamics simulation. The regulatory effects of GXⅡ on autophagy were confirmed by Western blotting and transmission electron microscopy in mouse hippocampus, and further verified in LPS-stimulated primary hippocampal neurons focusing on ATG3-mediated pathway.

GXⅡ treatment ameliorated dyslipidemia and reduced aortic plaque formation in mice with AS co-depression. It also demonstrated antidepressant effects in the OFT, SPT and TST. Network pharmacology analysis identified 77 overlapping targets among GXⅡ, AS and depression, with 15 core genes selected via PPI network topology. Molecular docking and dynamics simulation revealed robust binding interactions between GXⅡ constituents and AKT1, STAT3, and TLR4. Notably, GXⅡ restored the suppressed autophagy in the hippocampus of AS co-depressed mice, as evidenced by upregulated LC3, Beclin1 and ATG3 expression. These autophagy-regulating effects were consistently observed in primary hippocampal neurons and were reduced upon lentiviral knockdown of ATG3, confirming its essential role in mediating GXⅡ's therapeutic action.

GXⅡ ameliorates atherosclerosis and depression-like behaviors in comorbid mice via modulating the AKT1-STAT3-TLR4 signaling axis and activating ATG3-mediated autophagy. These findings provide pharmacological evidence to support the application of GXⅡ in the treatment of AS co-depression, and highlight ATG3 as a potential therapeutic target.