Abstract
Sipunculus nudus, an old marine species, has great potential for use as functional seafood due to its various bioactivities. Its potential antithrombotic activity pushed us to isolate the bio-active components bio-guided by tracking fibrinolytic activity. As a result, a novel protease named as SK (the kinase obtained from S. nudus) was obtained, which possessed a molecular weight of 28,003.67 Da and 15 N-terminal amino acid sequences of PFPVPDPFVWDTSFQ. SK exerted inhibitory effects on thrombus formation through improving the coagulation system with dose-effect relationship within a certain range. Furthermore, in most cases SK got obviously better effect than that of urokinase. With the help of untargeted mass spectrometry-based metabolomics profiling, arachidonic acid, sphingolipid, and nicotinate and nicotinamide mechanism pathways were found to be important pathways. They revealed that the effect mechanism of SK on common carotid arterial thrombosis induced by FeCl3 was achieved by inhibiting vessel contraction, platelet aggregation, adhesion, and release, correcting endothelial cell dysfunction and retarding process of thrombus formation. This study demonstrated SK was a promising thrombolytic agent on the basis of its comprehensive activities on thrombosis, and it should get further exploitation and utilization.
Highlights
Thrombotic disorders, including ischemic stroke, unstable angina, pulmonary embolism (PE), deep venous thrombosis (DVT) and myocardial infarction, are major causes of morbidity and mortality worldwide [1]
After the graded precipitation while using (NH4)2SO4, the crude protein was further purified by hydrophobic interaction chromatography that is based on its hydrophobicity
The results indicated that the RSD% of repeatability was between 3.06% and 8.15% and RSD% of stability was between 4.35% and 7.35%, which was at 0, 2, 4, 8, 12, and 24 h during a single day
Summary
Thrombotic disorders, including ischemic stroke, unstable angina, pulmonary embolism (PE), deep venous thrombosis (DVT) and myocardial infarction, are major causes of morbidity and mortality worldwide [1]. A number of antithrombotic agents, which include anticoagulant, antiplatelet, and fibrinolytic agents, are available for the treatment and prevention of thrombosis. The long-standing use of anticoagulant can cause a number of side effects, such as bleeding, thrombocytopenia, changes in lipid metabolism, and osteoporosis [2,3,4]. The antithrombotic agents, such as nattokinase, streptokinase, urokinase, and t-PA are exploited for treatment of thrombosis and they show significant therapeutic effects [5,6]. These antithrombotic agents are very expensive and still present some inescapable side effects [5,6,7]. Increasing studies indicate that many new thrombolytic agents are identified and characterized from natural resources, such as Eisenia foetida, snake venoms, Neanthes japonica, Urechis unicinctus, and so on [8,9,10]
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