中华硬蜱血小板集聚抑制剂的分离纯化与活性研究

Isolation and Purification of an Inhibitor on Platelet Aggregation from Ixodes sinensis

目的研究蜱类抗血小板集聚的生物活性成分,了解其与其宿主相互作用的分子机制。方法用葡聚糖Sephadex G-50凝胶过滤及高效液相从半饱吸血的中华硬蜱(Ixodes sinensis)唾液腺中分离纯化具有血小板集聚抑制活性的蛋白质,用飞行质谱测定该抑制剂的分子量,并用兔富血小板血浆研究其血小板集聚抑制活性。结果通过液相分离,从中华硬蜱唾液腺中得到了一种血小板集聚抑制剂,用飞行质谱测定该抑制剂的相对分子质量(Mr)为8 065。该抑制剂对二磷酸腺苷(ADP)诱导的血小板集聚表现强烈的抑制活性,在浓度为10μg/mL时,可以抑制90%以上的血小板集聚。结论首次分离获得中华硬蜱唾液腺血小板集聚抑制剂,该抑制剂对硬蜱顺利获取宿主血餐至关重要。

Objective To study the bioactive components in ticks which inhibit platelet aggregation, and to understand the molecular mechanism of tick-host interaction. Methods Sephadex G-50 gel filtration and high performance liquid chromatography （HPLC） were used to purify the platelet aggregation inhibitor from lxodes sinensis. Its molecular weight and purity were checked by matrix-assisted laser desorption ionization time-of-flight （MALDI-TOF） mass spectrometry. Platelet-rich plasma （PRP） of rabbit was used to examine the function of platelet aggregation inhibitor. Results A purified platelet aggregation inhibitor was identified from Lsinensis with a molecular weight of 8 065. It inhibited platelet aggregation induced by ADP with strong potency. The inhihition of platelet aggregation reached over 90% under a concentration of 10μg/ml. Conclusion An inhibitor of platelet aggregation from l.sinensis was identified, which may play an important role for ticks to successfully get blood meal from their hosts.