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目的:从地鳖虫中分离纯化得到纤溶活性蛋白(EFP),并研究其溶栓、溶血及对小鼠S180腹水瘤细胞的抑制作用。方法:通过硫酸铵分段盐析、DEAE-纤维素柱层析等方法从地鳖虫中分离纯化得到一种地鳖虫纤溶活性蛋白,对其进行了溶栓、溶血实验,采用MTT法和集落形成法研究了EFP对小鼠S180腹水瘤细胞的抑制作用。结果:分离得到的EFP分子量为87.2 kDa,具有一定的溶栓和溶血作用,并且其溶栓和溶血作用都具有浓度依赖性,低浓度(≤0.12mg/mL)的EFP不具有溶血作用。且EFP对S180腹水瘤细胞生长及集落形成都有抑制作用。结论:地鳖纤溶活性蛋白具有较强溶栓活性和微弱的溶血性,且对小鼠S180腹水瘤细胞有一定的抑制作用。
OBJECTIVE: To isolate and purify fibrinolytic active protein (EFP) from Eupolyphaga sinensis and investigate its inhibitory effect on thrombolysis, hemolysis and on mouse S180 ascites tumor cells. Methods: A fibrinolytic active protein from ground beetle was isolated and purified by segmented salting-out and DEAE-cellulose column chromatography. Thrombolytic and hemolysis experiments were carried out. MTT assay and colony Formation method to study the inhibitory effect of EFP on mouse S180 ascites tumor cells. Results: The molecular weight of the isolated EFP was 87.2 kDa. It had certain thrombolytic and hemolysis effects. The thrombolytic and hemolysis effects of EFP were concentration-dependent. EFPs with low concentration (≤0.12 mg / mL) had no hemolysis. And EFP on S180 ascites tumor cell growth and colony formation are inhibited. CONCLUSION: The fibrinolytic active protein from turtle has strong thrombolytic activity and weak hemolytic activity, and it has certain inhibitory effect on mouse S180 ascites tumor cells.