Preliminary Study On The Approach Of Anticoagulant Efficacy By Alkaline Extracting Jujube Polysaccharides

We have a rich resource of jujube in our province. In jujube, there are lots of polysaccharides that is proved to has multiple efficacies. The studies on the efficacies of jujube polysaccharides, however, are more focused in the fields of tissues and organs protection, fatigue resistance and oxidation,etc. The research of other efficacies and mechanism is necessary if we want to widen the applied range.Jujube has noticeable effect on promoting blood circulation, and the study on the activities of jujube polysaccharides is short. A lot of high performance antithrombotics, at the same time, are proved to have adverse reaction. For that reason, the study on the coagulation pathway is important.The experiments mainly reached following results:1. Different extraction methords of jujube polysaccharides had influences on their anticoagulant activities. It was better to extract with alkali, and showed significant positive correlation between anticoagulant activities and polysaccharides concentrations.2. The alcohol concentrations for deposition affected anticoagulant significantly, and 60%-80% was the best to obtain products with higher anticoagulant activities. The higher the concentrations was, the better; the activities had positive correlation with polysaccharide content and had negative correlation with protein content.3. Among the method of deproteinization, Sevage was the best for preparation. The APTT of the Sevage group was 1.32 times longer than blank control. Among the method of decoloration, activated carbon group was better and the APTT was 0.67 times longer than blank control.4. The study of coagulation pathway showed that jujube polysaccharides could not affect the number and the behaviour of blood platelet; the quality of thrombus in polysaccharide solution was decrease in 24 hours but less than blank control; the activities of coagulation factors had individual declines of 14.7%, 20.0% and 19.6% for the reason of jujube polysaccharides, the inhibitions were obviously. These showed that the anticoagulant activities were accomplished through the inhibition on coagulation factors.5. DEAE-52 cellulose column chromatography purification of jujube polysaccharides resulted in four constituents; collected them and studied the anticoagulant activities. The APTT of four constituents were 0、15.100、45.100 and 181.500. Component I and component Ⅱ showed the anxo-action on anticoagulant activities, component Ⅲ showed weak inhibitory action and component Ⅳ showed strong inhibitory action on anticoagulant activities. Sephacryl S-300 gel chromatography and UV scan showed that component Ⅱ was glucoprotein that was uniform, the range of molecular dimension was small, had the characteristic absorption of polysaccharide and protein but no pigment; component Ⅳ was polysaccharide that was uniform, the range of molecular dimension was bigger and had no protein or pigment.6. The inhibition ratios of component Ⅱ on the activities of three coagulation factors were 6.4%、6.8% and 4.1%; the inhibition ratios of component Ⅳ on the activities of three coagulation factors were 39.8%、 36.9% and 44.3%.7. Analyse monosaccharide composition, partial acid hydrolysis, malaprade reaction and Smith degradation by gas chromatography, the main monosaccharide composition of Ⅱ was Ara:Xyl:Gal=10.4:1:1.7; proportion of 1→3,1→2 and 1→6 glucosidicbondwas about 18.85:5.15:1;the main monosaccharide composition of Ⅱ was Xyl:Glc:Gal=6.7:1.8:1:1.6; proportion of 1→3,1 →2 and 1 →6 glucosidicbondwas about 5.48:1.09:1.8. Study the conformation by β-elimination, Iodine-potassium iodide reaction and Congo red reaction. Results of them showed that both of two components had O-glucosidicbond; they were both polysaccharides that was not belong to starch; the component Ⅱ might have ordered three helix structure and component Ⅳ might not.