# Shuxuening injection improves ventricular remodeling after myocardial infarction by indirectly promoting macrophage-mediated angiogenesis and lymphangiogenesis

## TL;DR

Shuxuening injection helps repair heart damage after heart attacks by boosting blood and lymph vessel growth through macrophages.

## Contribution

The study reveals that SXNI indirectly promotes angiogenesis and lymphangiogenesis via macrophage activation, identifying key active compounds.

## Key findings

- SXNI improves heart function and reduces infarct size in rats after myocardial infarction.
- SXNI activates VEGF-A/VEGFR2 and VEGF-C/VEGFR3 pathways, promoting angiogenesis and lymphangiogenesis.
- Ginkgolide A, B, rutin, and quercetin 3-neohesperidoside are key SXNI components that regulate VEGF expression in macrophages.

## Abstract

Macrophage-mediated angiogenesis and lymphangiogenesis after myocardial infarction (MI) are essential for restoring cardiac perfusion and lymphatic drainage, thereby limiting cardiac tissue ischemia, edema, and fibrosis. Shuxuening injection (SXNI) is commonly used in the treatment of cardiovascular diseases in clinical practice, but its mechanism of action in mitigating cardiac remodeling after MI is still unclear.

This study aimed to investigate the effect of SXNI on ventricular remodeling after MI and to clarify its mechanism of action.

SXNI were administered at doses of 1.05 (low-dose), 2.1 (clinical equivalent-dose), and 4.2 (high-dose) mL/kg/day over a 4-week period by using a rat MI model. Pharmacodynamic assessments encompassed cardiac function, infarct size, and fibrosis areas. Angiogenesis and lymphangiogenesis were assessed via immunohistochemical staining, western blotting, qRT-PCR, and ELISA. Assessment of cardiac edema and inflammatory status used gravimetry and ELISA. CCK-8, scratch wound, and tube formation assays were conducted to detect the direct and indirect (macrophage-mediated) effects of SXNI on HUVECs and SVEC4-10 cells. Western blotting examined the underlying mechanisms. The chemical composition of SXNI was determined by ultra-high performance liquid chromatography-Q exactive-mass spectrometry (UPLC-QE-MS). Key components of SXNI capable of binding to VEGF-A/VEGF-C were screened via molecular docking. The effects of these components on VEGF-A and VEGF-C levels in macrophages were then detected using qRT-PCR and ELISA.

In MI rats, SXNI significantly enhanced cardiac function, reduced infarct size, and suppressed cardiomyocyte apoptosis. SXNI activated the VEGF-A/VEGFR2 and VEGF-C/VEGFR3 signaling pathways, thereby promoting post-MI angiogenesis and lymphangiogenesis, consequently decreasing cardiac edema, inflammation, and dysfunction. Direct intervention with SXNI does not affect the viability or tube formation ability of HUVEC or SVEC4-10 cells. However, SXNI promoted the secretion of angiogenic factors by Raw264.7 cells. SXNI-induced macrophage stimulation indirectly enhanced the proliferation, migration, and tube formation of HUVECs and SVEC4-10 cells, thereby activating VEGFR2-mediated signals (AKT/ERK1/2) and VEGFR3-mediated signals (AKT/ERK1/2) in vitro, facilitating angiogenesis and lymphangiogenesis. Seventy chemical components were identified using UPLC-QE-MS mass spectrometry. Molecular docking results suggest that ginkgolide A, ginkgolide B, rutin and quercetin 3-neohesperidoside may bind to VEGF-A and VEGF-C proteins. Subsequent cellular experiments confirmed that these compounds could regulate the expression levels of VEGF-A and VEGF-C in macrophages.

SXNI activates macrophages to secrete VEGFs, such as VEGF-A and VEGF-C, which in turn activate the VEGFR2 and VEGFR3 signaling pathways in endothelial cells and lymphatic endothelial cells, thereby promoting angiogenesis and lymphangiogenesis and ameliorating ventricular remodeling. The key active ingredients in SXNI may be ginkgolide A, ginkgolide B, rutin and quercetin 3-neohesperidoside. This work provides valuable clinical evidence supporting its use in MI patient treatment.

The online version contains supplementary material available at 10.1186/s13020-026-01352-w.