Abstract
The biotechnological value of macroalgae for screening assays of thrombin generation-TG using sulfated polysaccharides-SPs as substitutes to heparin has been poorly explored. Five Brazilian species of macroalgae ( Gracilaria birdiae , Acanthophora muscoides , Halymenia sp., Caulerpa cupressoides and C. racemosa ) were analyzed and compared for their abundance, physical-chemical characteristics and in vitro anticoagulant assays of activated partial thromboplastin time-APTT, prothrombin time-PT and TG. Papain extraction yielded (p 100 kDa. These procedures, combined with the use of Stains-All, also indicated nonSPs. APTTs ranged from 2.81 ( A. muscoides ) to 21.30 IU ( Halymenia sp.) vs. heparin (193 IU), and were dependent on sulfation of the crude SPs. PT was not altered. With respect to TG assay, crude SPs modified concentration-dependent and independently from molecular mass TG by both intrinsic/extrinsic pathways in 60-fold diluted human plasma, with total intrinsic inactivation using crude SPs from A. muscoides in parallel to heparin (p < 0.05). Thrombosis in vitro is differentially modulated by distinct crude SPs from Brazilian seaweeds.
Highlights
Agardh were collected in September 2011 on seashore from the Flecheiras beach, Trairí, Ceará State, and they were taken to the Carbohydrates and Lectins Laboratory (CarboLec), Universidade Federal do Ceará, in plastic bags
The experimental analyses of the macroalgae crude sulfated polysaccharides (SPs) were performed at Connective Tissue Laboratory, Universidade Federal do Rio de Janeiro (UFRJ), Brazil
Samples of five different Brazilian seaweed species [G. birdiae, A. muscoides and Halymenia sp. (Rhodophyta); C. cupressoides var. lycopodium and C. racemosa (Chlorophyta)] collected in Flecheiras beach were digested with papain to obtain crude SPs and compare their respective physical-chemical characteristics by electrophoresis
Summary
In Brazil, a wide diversity of marine life (e.g., cucumbers, fishes and seaweeds) is distributed along the more than 8000 Km of coastline, revealing many biologically active metabolites (e.g., polysaccharides, proteins and carotenoids) which have already been isolated and studied as anticancer (Costa-Lotufo et al, 2006), anticoagulant, antithrombotic (Mourão et al, 2001; Fonseca et al, 2008; Rodrigues et al, 2012a) and anti-inflammatory (Pomin, 2012; Fernandes, Oliveira, & Valetin, 2014) agents, and as ingredients for food preparations (e.g., polysaccharides-based edible films) (Fenoradosoa et al, 2009; Paula et al, 2015). There is an increasing demand for seaweeds extracellular matrix complex sulfated polysaccharides (SPs) and the main biomaterials of economic value are agar and carrageenans, representing a billionaire market of over US$ 6 billion dollars year (Pereira & Costa-Lotufo, 2012) Regarding their chemical classes, Rhodophyceae and Phaephyceae are the most common sources of sulfated galactans and fucoidan or fucan, respectively (Pomin, 2012; Mourão, 2015), while sulfated heteropolysaccharides frequently occur in Chlorophyceae (Pomin & Mourão, 2008; Rodrigues et al, 2013; Arata et al, 2015). To obtain seaweeds SPs, enzyme-assisted extraction methods allow high yield and enhanced quality on the physical-chemical and biological properties of these molecules (Athukorala et al, 2006), considering the effort to obtain a commercial product on a large scale (Pereira & Costa-Lotufo, 2012)
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