中文摘要：

目的研究原矛头蝮蛇毒（PMV）及其组分作用于血液循环系统的部分生物学活性,进一步探讨其毒性机制。方法采用Sephadex G-75凝胶层析方法分离不同分子质量范围的蛋白组分FⅠ,FⅡ和FⅢ。贫血小板血浆调节富血小板血浆使血小板为3×1011L-1,血小板悬液分别与PMV,FⅠ,FⅡ和FⅢ0.03 g·L-1预孵育5 min,血小板聚集仪测定PMV及其组分诱导血小板聚集活性;PMV,FⅠ,FⅡ和FⅢ0.05 g·L-1分别与纤溶酶原0.1 U·L-1预孵育10 min,采用单一时间点测定和酶动力学测定PMV及其组分酶切发色底物S-2251的作用;将大鼠血浆与PMV,FⅠ,FⅡ和FⅢ1.0 g·L-1分别孵育5和30 min,测定凝血酶时间（TT）、活化部分凝血活酶时间（APTT）、凝血酶原时间（PT）和纤维蛋白原（FIB）水平;PMV,FⅠ,FⅡ和FⅢ10,50和250 mg·L-1处理微血管内皮细胞24 h,倒置相差显微镜观察细胞形态,MTT法检测细胞存活;PMV,FⅠ,FⅡ和FⅢ0.1 g·L-1与含卵磷脂和不含卵磷脂的豚鼠红细胞悬液分别孵育不同时间,计算溶血率。结果与对照组相比,PMV和高分子质量蛋白组分FⅠ（〉71 ku）诱导血小板聚集率显著升高〔（61.0±5.8）%和（56.9±5.9）%vs（12.4±4.1）%〕（P〈0.01）。酶切发色底物S-2251结果显示,PMV与中分子质量组分FⅡ（18~37 ku）具有酶切发色底物的作用（P〈0.01）。PMV和FⅠ可引起血浆凝固。与对照组相比,FⅡ组分和低分子质量蛋白组分FⅢ（〈10 ku）明显使TT,APTT和PT的时间延长（P〈0.01）。PMV,FⅠ及FⅡ导致内皮细胞解离悬浮,与对照组相比,细胞存活率下降,分别为（56.8±3.6）%,（71.6±3.8）%和（58.2±5.5）%。在无卵磷脂条件下,PMV和FⅡ可缓慢地引起红细胞轻度溶血;在卵磷脂参与下,PMV和FⅡ引起豚鼠红细胞急剧溶血,与正常对照组相比,在0.5 min内溶血率从（17.7±1.0）%分别升高至（81.0±4.0）%和（81.0±1.0）%（P〈0.01）。结论 PMV在体外表现出多方面的血?

英文摘要：

OBJECTIVE To investigate the effect of Protobothrops mucrosquamatus venom （PMV） and its fractions on functions of the circulatory system in vitro in order to better understand its toxicity mechanism. METHODS PMV was isolated to three fractions FⅠ, FⅡ and FⅢ with a different molecular mass range by Sephadex G-75 gel filtration chromatography. Platelet rich plasma was adjusted to 3＆#215;1011 L-1 by platelet poor plasma. Platelet suspension was incubated with PMV and its fractions 0.03 g.L-1 for 5 min, respectively, and platelet aggregation was determined on an LBY-NJ4 aggregometer. PMV and its fractions 0.05 g.L-1 were preincubated with plasminogen 0.1 U.L-1 for 10 min before chromogenic substrate cleavage activity was measured by endpoint and enzyme kinetics determination. PMV and its fractions 1.0 g.L-1 were incubated with rat plasma for 5 or 30 min, and thrombin time （TT）, activated partial thromboplastin time （APTT）, prothrombin time （PT） and fibrinogen （FlB） content were assayed. The microvascular endothelial cells were exposed to PMV and its fractions 10, 50 and 250 mg.L-1 , respectively, for 24 h, while the morphological change was observed using an inverted phase contrast microscope, and the cell viability was determined by MTT method. PMV and its fractions were incubated with guinea pig red blood cell suspension in the presence or absence of lecithin for different time, and hemolysis was measured. RESULTS Compared with normal control, platelet aggregation rate was significantly increased by PMV and FⅠ （＆gt;71 ku）〔（12.4±4.1）%,（61.0±5.8）% and （56.9±5.9）%〕（P〈0.01）. PMV and FⅡ （18-37 ku） significantly hydrolyzed chromogenic substrate S-2251（P〈0.01）. PMV and FⅠ caused plasma coagulation. Compared with normal control, FⅡ and FⅢ （〈10 ku） remarkably prolonged TT, APTT and PT（ P〈0.01）. Morphological observation revealed that PMV, FⅠ and FⅡdetached the adherent cells. Compared with normal control group, PMV, F Ⅰ and F ?