Lingguizhugan decoction inhibits the cleavage of LYVE-1 by MMP-9 and promotes lymphangiogenesis to improve myocardial infarction

Abstract

Background

Myocardial remodeling plays a crucial role in determining patient outcomes after myocardial infarction (MI). Emerging evidence from both preclinical and clinical studies highlights the beneficial effects of cardiac lymphangiogenesis in improving cardiac function and prognosis post-MI. Lingguizhugan decoction (LD), a traditional Chinese medicine, is extensively used in the treatment of ischemic heart disease. Nevertheless, its potential mechanisms are still not well understood.

Purpose

To determine whether LD can enhance post-MI myocardial remodeling by promoting lymphangiogenesis and to elucidate its molecular mechanisms

Methods

Surgical ligation of the left anterior descending artery (LAD) was utilized to establish MI rat model. Cardiac structure and function were assessed using histopathological staining and echocardiography. A transgenic zebrafish model was used to confirm that lymphangiogenesis plays a key role in LD’s cardioprotective effects. Network pharmacology analysis was conducted to predict the potential mechanisms underlying LD’s therapeutic action in MI. The expression levels of matrix metalloproteinase-9 (MMP-9) and lymphatic vessel endothelial receptor-1 (LYVE-1) were assessed in MI rats. Transcriptomic data mining from MI patients and in vitro protein interaction validation were conducted to explore the relationship between MMP-9 and LYVE-1. Key proteins involved in the interleukin-17 (IL-17) signaling pathway were analyzed using western blotting and qRT-PCR. Furthermore, mass spectrometry imaging was conducted to identify potential bioactive compounds in LD that regulate the IL-17 signaling pathway.

Results

LD significantly improved cardiac function and mitigated adverse myocardial remodeling in left anterior descending artery-ligated rats. Notably, LD promoted cardiac lymphangiogenesis and improved cardiac function in transgenic zebrafish treated with verapamil, with further validation using lymphangiogenesis inhibitors. Based on network pharmacology findings and previous studies, MMP-9 and LYVE-1 were identified as key targets of LD. LD decreased MMP-9 expression and increased LYVE-1 levels in MI rat hearts. A strong correlation was observed between MMP-9 and LYVE-1, suggesting a potential regulatory relationship. Additionally, LD downregulated key proteins involved in the IL-17 signaling pathway, indicating its role in modulating inflammatory responses. Finally, four biologically active compounds dehydrotrametenonic acid, ethyl p-methoxycinnamate, atractylon, and licoricone were identified in cardiac tissue as potential regulators of the IL-17R/MMP-9/LYVE-1 axis, contributing to LD’s lymphangiogenesis-promoting effects.

Conclusions

LD enhances post-MI ventricular remodeling by promoting lymphangiogenesis and modulating the IL-17R/MMP-9/LYVE-1 signaling pathway. Dehydrotrametenonic acid, ethyl p-methoxycinnamate, atractylon, and licoricone may serve as key bioactive compounds responsible for LD’s therapeutic effects.