Abstract
The structural complexity of the agaran type-sulfated polysaccharides (SPs) found in Acanthophora muscoides limits its investigation as anticoagulant alternative to heparin which induces clot complications. This study was extended to evaluate the properties of a SPs fraction and its alkali/desulfated derivatives on an intrinsic pathway-induced thrombin generation (TG) continuous model using 60-fold diluted normal or serpins-depleted human plasma. 0.75 M NaCl-eluted SPs fraction by DEAE-cellulose chromatography containing sulfate (35.20%), total sugars (55.97%) and no proteins showed charge homogeneity and heterogeneous molecular weight by agarose/polyacrylamide gel electrophoresis, respectively, using sequential staining with toluidine blue and Stains-All. Fourier Transform Infrared spectroscopy confirmed agaran-structure. Intact fraction poorly acted on the activated partial thromboplastin time (3.10 IU) than heparin (193 IU), but there was a preponderance of the serpin-independent effect than serpin-dependent one in TG assay comparing both systems was continually recorded. Heparin abolished plasma TG, but was inactive in depleted human plasma. While desulfated derivative of the respective fraction anticipated and induced thrombin formation vs. untreated plasma. The results suggested that sulfated sugars residues in the sacharide units of the polymer appear to be important to attenuate TG in vitro.
Highlights
The blood coagulation system is a complex sequence of enzymatic reactions orchestrated by coagulation factors
The current study investigated whether a sulfated polysaccharides (SPs) fraction isolated from A. muscoides and its chemically-modified derivatives inactive thrombin generation (TG) in 60-fold diluted depleted and/or normal human plasma by a continuous in vitro method
In this current study, employing anion-exchange (DEAE-cellulose) column for charged variously-fractions separation and electrophoresis in agarose or polyacrylamide gel for respective physical-chemical characterization confirmed that the abundance and the compositional profile of the SPs produced by A. muscoides would change according to the collection period (Quinderé et al, 2013; Rodrigues et al, 2016a)
Summary
The blood coagulation system is a complex sequence of enzymatic reactions orchestrated by coagulation factors. There are two pathways to its formation, the tissue factor-dependent extrinsic pathway and the contact factor-dependent intrinsic pathway, which are regulated by blood endogenous components, such as antithrombin, the most important physiological inhibitor of the coagulation (Rau, Beaulieu, Huntington, & Church, 2007). These pathways are explored by the activated partial thromboplastin time (APTT) and the prothrombin time tests, respectively, but they are of limited value and reflect only the first traces of the amount of thrombin formed. These thrombin generation (TG)-based coagulation assays aforementioned have been used as global tools for the study of plasma coagulability (Castoldi & Rosing, 2011; Duarte, Ferreira, Rios, Reis, & Carvalho, 2017) and analysis of anticoagulant agents
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