白树花多糖硫酸酯的抗凝血活性研究

Study on Anti-coagulation Activity of Polyporus Albicans Teng Sulfate

目的研究白树花多糖硫酸酯的抗凝血效果及抗凝血机理,为制备合成高效、安全的多糖硫酸酯抗凝剂提供实验依据。方法将可食用的白树花真菌进行发酵,提取水溶性多糖,采用氯磺酸-吡啶法进行硫酸酯化修饰,获得水溶性的白树花多糖硫酸酯。通过测定APTT、TT、凝血因子活性对白树花硫酸酯的抗凝血效果及抗凝血机理进行评估和探讨。结果对正常人血浆进行的抗凝血实验表明,白树花多糖硫酸酯具有明显的抗凝血活性,在5 mg/L的浓度下即可发挥效果,在10 mg/L浓度时,相当于150 U肝素的抗凝血效果;抗凝血机理研究结果表明,白树花多糖硫酸酯作用于内源凝血系统,主要通过抗凝血酶Ⅲ抑制凝血因子Ⅱa和Xa的活性发挥抗凝血作用,抗凝血机理与肝素完全相同。结论通过上述研究结果,我们认为白树花多糖硫酸酯具备肝素的抗凝血特性。同时由于是可食用真菌多糖的制备产物,在防生物污染方面具备一定优势。因此,白树花多糖硫酸酯可作为理想的肝素替代品进行开发及临床应用。

Objective To synthesize effective safe polysaccharide sulfate as anticoagulant,the anticoagulant activity and the anticoagulant mechanism of Polyporus Albicans Teng Sulfate（PATS） were investigated.Methods We extracted water-soluble polysaccharide from fermented mycelium of edible polyporus albicans（Imaz.） Teng,and got the water-soluble polyporus albicans teng sulfate（PATS） by modifing the water-solubility polyose with method of chlorosulfonic acid-pyridine.To evaluate the anticoagulant effect and study the anticoagulant mechanism of PATS,APTT,TT and the activity of coagulant factors were detected.Results The anticoagulant test in vitro with normal human plasma indicated that PATS has conspicuous anticoagulant activity.The dose could be as low as 5 mg/L to start anticoagulation activity.And at the dose of 10 mg/L,the effect was equivalence to that of heparin about 150U.Our study on anticoagulation mechanism suggested that PATS taking pathway of internal anticoagulant system.The anticoagulation activity of PATS was due to the inhibition of the coagulation factors Ⅱa Xa activities mediated by antithrombin AT-Ⅲ.This anticoagulation mechanism is absolutely identical to that of heparin.Conclusion We suggested that PATS have the similar anticoagulation characteristic to heparin.Meanwhile,being the production of edible fungus-polysaccharide,PATS has advantage on bio-safety.Therefore,PATS can be developed and used clinically as an ideal substitute for heparin.