Abstract
Marine and salmon polar lipids (PLs) extracted by conventional extractions with non-food-grade solvents (CE-salmon-PLs) possess antithrombotic bioactivities against platelet-activating factor (PAF) and thrombin. Similar effects of food-grade-extracted (FGE) marine PLs have not yet been reported. In this study, food-grade solvents were used to extract PLs from Irish organic farmed salmon (Salmo salar) fillets (FGE-salmon-PLs), while their antithrombotic bioactivities were assessed in human platelets induced by platelet aggregation agonists (PAF/thrombin). FGE-salmon-PLs were further separated by thin layer chromatography (TLC) into lipid subclasses, and the antithrombotic bioactivities of each subclass were also assessed. LC-MS was utilized to elucidate the structure-activity relationships. FGE-salmon-PLs strongly inhibited PAF-induced platelet aggregation, while their relevant anti-thrombin effects were at least three times more potent than the previously reported activities of CE-salmon-PLs. TLC-derived lipid fractions corresponding to phosphatidylcholines (PC) and phosphatidylethanolamines (PE) were the most bioactive lipid subclasses obtained, especially against thrombin. Their LC-MS analysis elucidated that they are diacyl- or alkyl-acyl- PC and PE moieties baring ω3 polyunsaturated fatty acids (PUFA) at their sn-2 position, such as eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA). Our results concerning the potent antithrombotic effects of FGE-salmon-PLs against both PAF and thrombin pathways strongly suggest that such food-grade extracts are putative candidates for the development of novel cardioprotective supplements and nutraceuticals.
Highlights
Marine polar lipids (PLs) have exhibited several beneficial activities against inflammation, thrombosis, and related disorders [1,2,3,4]
Total lipids (TL) of all samples (n = 6) of salmon fillets from the same batch of farmed salmon were extracted using food-grade solvents according to EU legislation for extracting fish oils and further separated into neutral lipids (NL) and PLs by a counter-current distribution, using food-grade solvents
Dietary supplementation of fish oil (MaxEPA) rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) for six weeks resulted in the reduction of platelet-activating factor (PAF) synthesis in healthy subjects, while PAF generation was reduced in monocyte monolayers in the EPA treatment group only, while PAF synthesis was increased by arachidonic acid (AA) [35]
Summary
Marine polar lipids (PLs) have exhibited several beneficial activities against inflammation, thrombosis, and related disorders [1,2,3,4]. Mar. Drugs 2019, 17, 62 the activities of several plasma membrane receptors related to important pathways of inflammation and thrombosis [1,2,3,4,5], such as the receptor for platelet-activating factor (PAF) [4,5,6]. Thrombin is a serine protease that participates in coagulant catalytic processes, such as the conversion of fibrinogen into fibrin and the activation of the V, VIII, XI, and XIII coagulation factors [7,8], as well as the activation of many cell types and platelets including platelet aggregation [7] Such soluble platelet agonists are produced during coagulation (e.g., thrombin) and inflammation (e.g., PAF) and play a critical role via G-protein-coupled receptors (GPCR) in platelet activation and thrombus formation [7]. These mediators present in high concentrations in atherosclerotic plaques and atherothrombotic events [6,8]
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