Combined Proteomics and Metabolomics Analysis of Pathological Mechanisms in Mouse Models of Coronary Heart Disease

Abstract

BACKGROUND: The pathogenesis of coronary heart disease is complex. A single omics approach is limited in elucidating its biological pathways, whereas multi-omics integration helps reveal molecular interaction networks across different levels, addressing the limitations of single-omics methods. OBJECTIVE: To investigate the pathological mechanisms of coronary heart disease in a mouse model using proteomics and metabolomics. METHODS: Healthy SPF-grade 8-week-old male C57BL/6 mice were randomly divided into a sham operation group and a model group. The mouse model of coronary heart disease was established by ligation of the left anterior descending coronary artery, while the sham operation group underwent threading without ligation. At 28 days post-surgery, cardiac function was assessed by echocardiography, and myocardial infarct size was evaluated by TTC staining. Ultra-high-performance liquid chromatography-tandem mass spectrometry was used to screen differentially expressed proteins and metabolites between groups, followed by integrated omics analysis. RESULTS AND CONCLUSION: Compared with the sham group, the model group exhibited reduced cardiac function, with significantly decreased left ventricular ejection fraction and left ventricular fractional shortening (P < 0.05), and significantly increased myocardial infarct size (P < 0.01). Proteomics identified 420 differentially expressed proteins, including 282 upregulated (e.g., Serum amyloid A protein, protein kinase D) and 138 downregulated (e.g., Protein YIPF5, E3 ubiquitin-protein ligase). KEGG pathway enrichment revealed involvement in ATP-dependent chromatin remodeling and renin-angiotensin system pathways. Metabolomics identified 155 differential metabolites, including 56 upregulated (e.g., Thromboxane, Tromethamine) and 99 downregulated (e.g., N-Acetyl-D-Tryptophan, D-Xylulose 5-Phosphate). KEGG analysis linked these to purine metabolism and glycerophospholipid metabolism. Integrated analysis found correlations between 26 differentially expressed proteins and 16 differential metabolites, involving proteins such as ATP1A3 and Hexokinase, and metabolites such as Cytochalasin B and Gluconasturtiin. CONCLUSION: The pathological mechanisms of coronary heart disease are closely related to disturbances in energy metabolism networks, activation of inflammatory-coagulation cascades, and dysregulation of ion homeostasis.

1. Introduction

Proteomics and metabolomics, as core technologies of systems biology, provide a new paradigm for deciphering complex life phenomena through their dynamic, holistic, and systematic research characteristics [1-2]. Proteomics comprehensively captures the expression profiles, post-translational modifications, and interaction networks of proteins in organisms, revealing the regulatory mechanisms of key proteins during disease development. Metabolomics precisely detects changes in endogenous metabolites in biological fluids or tissues, reflecting the dynamic imbalance of metabolic networks. The combined application of these two approaches enables elucidation of the pathophysiological processes of diseases from the 'protein-metabolite' dimension, which aligns closely with the holistic view and syndrome differentiation philosophy of traditional Chinese medicine.

Coronary atherosclerotic heart disease (coronary heart disease, CHD), caused by atherosclerotic lesions leading to stenosis or occlusion of coronary arteries, results in myocardial ischemia, hypoxia, or necrosis. It has become one of the leading causes of death and disability worldwide [3]. In traditional Chinese medicine theory, CHD falls under the categories of 'chest bi' and 'true heart pain', with blood stasis as its core pathological feature, and blood stasis syndrome is one of the most common clinical syndromes [4]. However, as a nonlinear complex system, the biological connotation of syndromes requires research methods adapted to its complexity [5-6].

Based on this, to deeply explore the pathological mechanisms of CHD in mouse models and identify key signaling pathways and potential biomarkers, this study employed ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to integrate proteomics and metabolomics for combined analysis. The aim is to provide in-depth data support and theoretical basis for elucidating the traditional Chinese medicine pathogenesis of CHD at the molecular level, and to open new research avenues for syndrome differentiation, efficacy evaluation, and new drug development for CHD.