Therapeutic effect and mechanism of danshensu on coronary heart disease using liquid chromatography combined with mass spectrometry metabolomics

Highlights

• Metabolic profile and biomarkers were discovered by comprehensive metabolome analysis.

• The pharmacological effect and mechanism of action of danshensu treatment was also revealed.

• A total of 26 potential biomarkers were screened out, and involving 8 metabolic pathways.

• DSS could significantly regulate the primary bile acid, glycerophospholipid, and lipid metabolism.

• This study dissects pharmacological effects and mechanisms of natural products via metabolomics.

Abstract

Metabolomics can discover the biomarkers and metabolic pathways, provides the possibility for insights into the pharmacological action and mechanism of natural products. The therapeutic effect and mechanism of danshensu (DSS) on total metabolic pathways has not been well investigated. The aim of this study was to explore the disturbed endogenous biomarkers and metabolic pathways reflecting the pharmacological activity of DSS, and mechanism of action of DSS using comprehensive metabolome analysis based on high-throughput metabolomics technology combined with ultra-high performance liquid chromatography (UPLC) coupled with quadrupole tandem time-of-flight mass spectrometry (Q-TOF-MS) and pattern recognition method. Through the changes of the overall metabolic profile and the related biomarkers, the intervention effect of natural product danshensu (DSS) treatment on CHD model rats was revealed. The results showed that after the model replication was established, the metabolic profile was clearly separated, and a total of 26 potential biomarkers were screened out, and involving 8 metabolic pathways. After different doses of DSS solution were given, a total of 20 biomarkers could be significantly regulated, mainly involving primary bile acid biosynthesis, glycerophospholipid metabolism, and lipid metabolism. It showed UPLC-MS-based metabolomics can be used for discovering potential biomarkers and metabolic pathways of CHD, and to further understand and dissecting pharmacological effects and mechanisms of natural products via metabolomics techniques.

Introduction

Coronary heart disease (CHD) is a kind of cardiopathy evocable by atherosclerotic lesion of left and right coronary arteries and their branches, which can cause vascular lumen obstruction, myocardial ischemia and even necrosis and other diseases [1]. It is a common disease among the middle-aged and the elderly, with extremely high morbidity and disability. In recent years, the onset age of the disease is getting younger and younger, which is mainly related to dangerous factors such as hyperlipidemia, diabetes, obesity, smoking and hypertension [2]. The main feature of CHD is the formation of arterial plaques, which narrow the coronary artery lumen and lead to paroxysmal or persistent angina pectoris [3]. Rupture of plaque leads to thrombosis, myocardial infarction and death due to cessation of blood flow [4]. Therefore, the theory of inflammation is very important in the etiopathogenesis of CHD. A lot of studies have shown that inflammatory factors such as interleukin-6, C-reactive protein (CRP), CD40 and myeloperoxidase (MPO) can be used for clinical diagnosis and prognosis observation of CHD [5], [6]. However, the existing methods are all performed after pathological changes occur, and there is no sensitive and accurate method for clinical early diagnosis of CHD. Metabolomics provides a new method for pre-disease diagnosis and drug therapy monitoring of CHD [7], [8].

Metabolism is the basic form of life activities and is a means of systematic research to analyze all metabolic changes of organisms after being stimulated by interference [9]. Investigating the metabolism in vivo under the disease state and studying the metabolic changes of endogenous substances induced by it can help people to preferable comprehend the pathogenesis, discover biomarkers of the illness and further guide the treatment [10]. At present, metabolomics technology has been applied in many fields such as drug research [11] and development, botany [12], and has shown a wide application prospect in clinical medicine including cardiovascular disease research [13]. Feng et al. [14] discovered GlcNAc-6-P, mannitol and 15 kinds of serum choline as potential biomarkers of CHD using metabolomics techniques. In recent years, UPLC-MS technology has become an important means of metabolomics research [15], [16]. UPLC ensures rapid separation with high resolution under high column pressure, while Q-TOF-MS not only has accurate molecular mass and high resolution of time-of-flight mass spectrometry, but also can obtain secondary mass spectrometry through collision and fragmentation again to accurately identify molecular structure [17]. Therefore, it is often used for accurate identification and quantitative analysis of metabolic markers in metabolomics [18], [19]. Metabolomics with UPLC-MS technology not only provides an efficient technical means for the clinical diagnosis and prevention of diseases[20], [21].

The herbal medicine has unique functions in preventing and protecting cardiovascular diseases [22], [23]. Compound Danshen Dropping Pills is a TCM prepared by extracting the effective components of traditional compound Danshen tablets. It has been extensive used in clinical cure of CHD, angina pectoris and other diseases with chest tightness symptoms[24], [25]. Its main water-soluble component danshensu (DSS, salvianic acid A) has pharmacological effects of anti-inflammation[26], [27], protecting myocardium[28], inhibition of thrombosis, anti-liver fibrosis[29], anti-cancer[30] and immunity improvement[31], [32]. In this study, UPLC-MS-based combined with metabolomics technology was used to explore the potential biomarkers and related metabolic pathways of CHD. Based on the establishment of the model, the therapeutic effect and potential therapeutic targets of DSS on CHD are evaluated to provide evidence for the effectiveness of DSS in treating CHD.