Preliminary study on the anticoagulation mechanism of glycosaminoglycan from Urechis unicinctus
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摘要: 采用人乏AT-Ⅲ血浆、乏HCII血浆及两种肝素辅因子均缺乏的血浆,探讨单环刺螠糖胺聚糖的抗凝血途径;通过纠正乏因子血浆所致的凝固时间延长的能力测定人血浆凝血因子Ⅱ、Ⅴ、Ⅶ、Ⅷ、Ⅸ、Ⅹ、Ⅺ、Ⅻ的活性,凝固法测定纤维蛋白原浓度;体外复钙凝血实验,研究糖胺聚糖对复钙凝血时间的影响;甲基百里酚蓝比色法测定大鼠和小鼠血清中钙离子浓度。结果表明:单环刺螠糖胺聚糖的抗凝血酶作用主要呈AT-Ⅲ依赖性,并能够钝化肝素辅因子Ⅱ抑制凝血酶的活性;单环刺螠糖胺聚糖显著降低血浆凝血因子Ⅴ、Ⅶ、Ⅹ、Ⅷ、Ⅸ、Ⅺ、Ⅻ的活性（p<0.05）;糖胺聚糖可通过屏蔽血液中Ca2+,延长大鼠血复钙凝血时间,极显著降低大鼠和小鼠血液中的钙离子浓度（p<0.01）,且其降低血液钙离子浓度的作用极显著优于肝素钠（p<0.01）。Abstract: In this paper, the anticoagulation approaches of glycosaminoglycan was studied with AT-III-deficient plasma, HC-II-deficient plasma and both factor-deficient plasma in human. The activities of coagulation factor II, V, VII, VIII, IX, X, XI, XII and the fibrinogen content in human plasma were measured using the solidification method. The influence of glycosaminogly could on blood recalcification time in rats was studied by using recalcification experiment. The Ca2 +concentrations in rats and mice serum were measured through methyl thymol blue colorimetric method. The results showed that the antithrombin role of glycosaminoglycan from Urechis unicinctus mainly presented AT- Ⅲ dependency. Glycosaminoglycan could passivate the thrombin inhibitory activity of heparin cofactor II. Glycosaminoglycan significantly reduced the activities of coagulation factor V, VII, X, VIII, IX, XI, XII in the plasma ( p < 0. 05) . Besides, glycosaminoglycan could extend prolong recalcification time in rats by shielding Ca2+in plasma, and significantly reduced Ca2+concentration in rats and mice serum (p<0.01) . Glycosaminoglycan reduced the Ca2+concentration in a more intensive way than that of heparin sodium (p<0.01) .
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Keywords:
- Urechis unicinctus /
- glycosaminoglycan /
- anticoagulation mechanism
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