蝮蛇毒抗凝蛋白组分对人血管内皮细胞EC-304作用的实验研究

Research on Function of the Anti-clotting Protein Component from Venom of Agkistrodonon to Human Vascular Endothelial EC-304 cell

目的从皖南尖吻蝮蛇毒中分离纯化出抗凝蛋白组分,观察其对人血管内皮细胞的抑制作用。方法体外培养人血管内皮细胞EC-304,CCK-8比色法测定蝮蛇毒抗凝蛋白组分对体外培养的EC-304细胞的细胞毒作用,相差显微镜观察EC-304细胞经蝮蛇毒抗凝蛋白组分处理后,细胞形态学、生物化学等方面的变化。结果蝮蛇毒抗凝蛋白组分对EC-304细胞具有明显的生长抑制作用（P〈0.05）,浓度为1μg/mL的蝮蛇毒抗凝蛋白组分对EC-304细胞即有很强的抑制作用,随着剂量的加大,其抑制率不断上升;经蝮蛇毒抗凝蛋白组分处理后的EC-304细胞发生形态学变化,显微镜下观察到细胞圆缩,悬浮细胞增多,细胞分裂相减少,培养液内细胞碎片明显增多。结论皖南尖吻蝮蛇毒抗凝蛋白组分对EC-304细胞具有很强的抑制作用,诱导EC-304细胞发生凋亡可能是其机制之一。

Objective To investigate the effects of the anti-clotting protein component from venom of Agkistrodonon on human Vascular endothelial EC-304 cell.Methods CCK-8.Method was used to detect the antitumor effect of EC-304 in vitro;Phase-contrast-microscop was used d to observe the alteration of cell form,biochemistry variety of the Processed cells.Results The generation depressant effects of EC-304 cells culturedin vitro were detected by CCK-8 method（P0.05）,The 1μg/mL dosage the anti-clotting protein component from venom of Agkistrodonon had a very strong inhibition to EC-304 cells,and there were dosage-time dependent relationships.The suspending cells and the cellular debris got more,the process of cell division got less.Conclusions The anti-clotting protein component from venom of Agkistrodonon has very strong inhibition to the EC-304 cells.Besides the direct damaging effect,inducing apoptosis is one of its functions.