Antiplatelet Aggregation Activity Of The Sorghum Vinegar Extract

The aberrant activation of platelets plays essential role in the pathogenesis of physiological hemostatic and pathological thrombotic processes,which are commonly associated with cardiovascular diseases.Sorghum vinegar is an excellent source of physiological substances with multiple health benefits,these substances include organic acids,saccharides and amino acids.We hypothesized that the ethanol(60%,v/v)extract of sorghum vinegar may inhibit platelet aggregation.In this study,the sorghum vinegar extract(SVE)was first evaluated for antiplatelet activity.Also the mechanism of monosaccharides and alditols antiplatelet activity was demonstrated by enzyme-linked immunoassay method and computational docking study.The main results are as follows:The crude sorghum vinegar extract was collected by ultrasonic assisted solvent extraction.AB-8 macroporous resin column(50 cm×2 cm)was used for the separation and purification of crude extract.Desorption solution was used to remove any ethanol by rotary evaporation,followed by freeze-drying.The dried extract was named SVE and stored at-18? for further study.Compounds in SVE were identified based on the retention times and mass spectra of gas chromatography,and quantified based on the peak areas of the identified compounds.A total of 25 peaks were identified,which amounted to 52.63%of the total SVE.Monosaccharides accounted for 44.24%of SVE,alditols accounted for 6.77%,compounds 3,4-(methylenedioxy)mandelic acid(0.27%)and ethylglucoside(1.35%)were also present.Monosaccharides and alditols accounted for 51.01%of the SVE,this suggested that monosaccharides and alditols are the main compounds of SVE.Antiplatelet activity of SVE was examined using the optics turbidimetric method.SVE showed significant anti platelet activity induced by arachidonic acid(AA),collagen,adenosine diphosphate(ADP)and thrombin in a concentration-dependent manner(p<0.05).SVE(0.50 mg/mL)reduced the AA-induced aggregation rate to 10.35%±0.46%,which was comparable to acetylsalicylic acid(aspirin,ASA)(0.50 mg/mL,6.35%±0.58%),a medical standard.The result suggested that SVE could be a multiple-step inhibitor of platelet aggregation,especially inhibit AA-induced platelet aggregation.To determine the inhibitory activity of SVE on cyclooxygenase-1(COX1),thromboxane-A2 synthase(TXS)and thromboxane-A2(TXA2)production experiments were conducted using the COX1,TXS and TXB2 EIA kit.The results showed that SVE(0.10,0.25,0.50 mg/mL)significantly inhibited the activity of COX1 and TXS(p<0.05).TXB2 production induced by AA(100 ?M)was significantly inhibited by SVE(p<0.05).Computational docking study showed that alditols(L-erythritol,L-arabitol,xylitol and D-sorbitol),monosaccharides(D-glucopyranose,D-fructofuranonse,D-xylopyranose,D-galactopyranose and D-ribose),ethyl glucoside and 3,4-(methylenedioxy)mandelic acid could dock directly into the active site of COX1.In conclusion,this study was the first to show that SVE,which is composed mainly of alditols and monosaccharides,has considerable antiplatelet activity.SVE could be a multiple-step inhibitor of platelet aggregation.The inhibitory activity of SVE is associated with the AA metabolic pathway,and alditols and monosaccharides could be major bioactive compounds based on the computational docking results.Alditols and monosaccharides in sorghum vinegar have a therapeutic advantage with respect to preventing cardiovascular disease because of their potential to inhibit multiple steps in the platelet aggregation pathway.