三种蝮蛇抗栓酶注射剂对大鼠和家兔血小板聚集的影响

用比浊法研究三种蝮蛇抗栓酶注射剂对大鼠和家兔血小板聚集的影响,结果表明蝮蛇抗栓酶注射剂在试管内浓度0.045u/ml 时对大鼠和家兔血小板聚集无明显抑制作用。静脉注射蝮蛇抗栓酶注射剂1u/kg 对家兔血小板聚集有明显抑制作用。尤其是蛇岛蝮蛇抗栓酶注射剂抑制作用最为显著。     

精制蝮蛇抗栓酶治疗冠心病心绞痛46例疗效观察

精制蝮蛇抗栓酶是蛇毒抗栓酶的第三代制剂,具有抗凝、溶栓、去纤、抗血小板粘附聚集、阵脂、扩张血管、改善微循环等多种功能。我院1992～1993年应用精制蝮蛇抗栓酶治疗冠心病心绞痛46例,取得良好疗效,报道如下。 临床资料 一、一般资料 本文按治疗方法不同分为两组,1组应用精制蝮蛇抗栓酶,从1992年1月起,至1993年3月收住院冠心病心绞痛患者中随机     

应用精制蝮蛇抗栓酶治疗脑梗塞133例总结

精制蝮蛇抗栓酶是低毒复合酶制剂,具有抗凝溶栓、去纤、抗血小板粘附聚集、降脂、扩张血管和促进神经细胞恢复等多种功能。我院自1990年以来,应用大连司威特制药有限公司生产的精制蝮蛇抗栓酶治疗600余例,现总结如下。     

三种蝮蛇抗栓酶的毒性及性质的实验研究

三种蝮蛇抗栓酶的毒性强度用LD_(50)表示,分别是,蛇岛蝮蛇抗栓酶为1.68±0.35u/kg,东北陆生白眉蝮蛇抗栓酶为5.23±0.61u/kg,浙江蝮蛇抗栓酶为1.94±0.15u/kg。对家兔亚急性毒性实验表明,对血细胞、肝及肾功能、脏器等无明显影响。蛇岛蝮蛇抗栓酶可减少血小板的数量。东北陆生白眉蝮蛇抗栓酶和浙江蝮蛇抗栓酶具有体外血浆凝固性。蛇岛蝮蛇抗栓酶、东北陆生白眉蝮蛇抗栓酶表现了以血循环毒为主的毒性,浙江蝮蛇抗栓酶表现了以神经毒为主的毒性。     

合理使用蝮蛇抗栓酶

合理使用蝮蛇抗栓酶山东烟台毓璜顶医院（２６４０００）李秀芹蝮蛇抗栓酶是从蝮蛇毒中提取的一种酶制剂，经临床观察，该药具有抗凝、溶栓、去纤、抗血小板粘附聚集、降低血液粘度，改善微循环、扩张血管、促进神经细胞恢复等多种功效。临床上主要用于治疗脑血栓、冠心病...     

蝮蛇抗栓酶的临床毒副反应

蝮蛇抗栓酶的临床毒副反应陈伟，郭健琳（乌鲁木齐铁路中心医院内科乌鲁木齐８３００１１）蝮蛇抗栓酶（Ｓｖａｔｅ）是一种生物酶类制剂。它含有近５０种毒素蛋白、酶及少量其它蛋白质，属异种蛋白；其主要作用是可消耗纤维蛋白原，降低血粘度，抑制血小板聚集，同时具有...     

蝮蛇抗栓酶治疗心血管疾病40例

应用蝮蛇抗栓酶(由江浙蝮蛇毒中提取)治疗心血管疾病40例,剂量为0.01-0.02U/kg,稀释后静滴,qd,21d为一疗程。治疗后患者的全血和血浆粘度、血小板聚集率、纤维蛋白原含量和血脂浓度均有显著下降。另有类似的40例患者用曲克芦丁400mg/d,静滴,21d为一疗程作对照,结果发现蝮蛇抗栓酶组(包括临床疗效)均明显优于对照组,且不良反应轻微,未发现肝、肾功能障碍和明显出血倾向。     

精制蝮蛇抗栓酶致频发房性早搏1例

精制蝮蛇抗栓酶致频发房性早搏１例毕波（内蒙古自治区通辽市中医院，通辽０２８０００）关键词蛇毒类；期外收缩；脑梗死蝮蛇抗栓酶具有抗凝、溶栓、去纤、抗血小板粘附聚集、降脂、扩张血管、改善微循环等多种功效［１，２］，现已用于治疗闭塞性心、脑血管疾病。常见的...     

肝素与蝮蛇抗栓酶治疗高粘血症疗效对比分析

肝素与蝮蛇抗栓酶均有抗凝、抑制血小板聚集的作用,故临床上均可用于高粘血症的治疗,其中以后者应用较广泛。我们于1993年10月以来,开展用肝素治疗高粘血症,并与蝮蛇抗酸酶治疗该病的疗效加以对比,现分析报告如下。 资料与方法     

蝮蛇抗栓酶对脑梗塞患者血小板数量的影响与临床关系

蝮蛇抗栓酶对脑梗塞患者血小板数量的影响与临床关系山西医学院第一附属医院（０３０００１）张生林，刘玉玺，马晋萍山西省汽车制造厂贾殿华蝮蛇抗栓酶治疗急性脑梗塞颇有疗效［１，２］，但在用药过程中患者有可能会出现疲乏、肢体无力等症状。我们观察到这些症状和血小板数量减少有关，现将其结果报告如下。临床资料２２例入院的急性脑梗塞患者，全部作了ＣＴ扫描。其中男性１５例，女性７例；年龄为１６～７０岁。入院后行一般常规神经系统检查，用专门表格记录用药前后体征、症状与血小板数量的变化。药前行二次血小板计数，取其均值作为对照组。药后分别于３、６、９天及两周时取耳垂毛细血管测血小板数。若患者在用药过程中出现皮疹、乏力时则每天测血小板数。当血小板下降到６０～８０×１０９／Ｌ时或疲乏、肢体无力明显时即停药，待症状好转或消失测血小板数，行第二个疗程治疗。蝮蛇抗栓酶为沈阳第一制药厂生产。开始用量为０．５个酶活力单位，第一个疗程每隔３天增力０．２５个单位，最大量达１．７５个单位。结果一、应用蝮蛇抗栓酶后不同时期内血小板数量的变化见表１。二、血小板变化与临床症状与体征间关系。用蝮蛇抗栓酶后，部分患者会出现搔痒、皮肤出血点、疲乏、肢体乏力和不适等     

Antithrombotic effect of a protein-type I class snake venom metalloproteinase, kistomin, is mediated by affecting glycoprotein Ib-von Willebrand factor interaction

Binding of von Willebrand factor (vWF) to platelet glycoprotein (GP) Ib-IX-V mediates platelet activation in the early stage of thrombus formation. Kistomin, a snake venom metalloproteinase (SVMP) purified from venom of Calloselasma rhodostoma, has been shown to inhibit vWF-induced platelet aggregation. However, its action mechanism, structure-function relationship, and in vivo antithrombotic effects are still largely unknown. In the present study, cDNA encoding kistomin precursor was cloned and revealed that kistomin is a P-I class SVMP with only a proteinase domain. Further analysis indicated that kistomin specifically inhibited vWF-induced platelet aggregation through binding and cleavage of platelet GPIbalpha and vWF. Cleavage of platelet GPIbalpha by kistomin resulted in release of 45- and 130-kDa soluble fragments, indicating that kistomin cleaves GPIbalpha at two distinct sites. In parallel, cleavage of vWF by kistomin also resulted in the formation of low-molecular-mass multimers of vWF. In ex vivo and in vivo studies, kistomin cleaved platelet GPIbalpha in whole blood. Moreover, GPIbalpha agonist-induced platelet aggregation ex vivo was inhibited, and tail-bleeding time was prolonged in mice administered kistomin intravenously. Kistomin's in vivo antithrombotic effect was also evidenced by prolonging the occlusion time in mesenteric microvessels of mice. In conclusion, kistomin, a P-I class metalloproteinase, has a relative specificity for GPIbalpha and vWF and its proteolytic activity on GPIbalpha-vWF is responsible for its antithrombotic activity both in vitro and in vivo. Kistomin can be useful as a tool for studying metalloproteinase-substrate interactions and has a potential being developed as an antithrombotic agent.     

短尾蝮蛇毒磷脂结合抗凝蛋白对动物血栓的抗栓作用

目的从短尾蝮蛇毒中分离纯化磷脂结合抗凝蛋白(PBAP),研究其对动物血栓的抗栓作用。方法用Chandler法制备家兔体外血栓和半体内血栓模型;用Reyers法制备大鼠静脉血栓模型;用Nowork法制备家兔肺动脉栓塞模型。测定PBAP对家兔体外血栓、半体内血栓、肺动脉栓塞和大白鼠静脉血栓的抗栓效应。结果短尾蝮蛇毒PBAP对家兔体外血栓、半体内血栓、肺动脉栓塞和大白鼠静脉血栓干湿重均有明显减轻作用,与生理盐水对照组比较,差异均有统计学意义(P<0.05或P<0.01)。结论PBAP抗凝蛋白对动物血栓具有明显的抗栓作用,具有潜在的临床应用价值。     

Inhibition of human platelet aggregation by L-amino acid oxidase purified from Naja naja kaouthia venom.

L-Amino acid oxidase (LAO) widely exists in snake venoms. Purification of LAO from the Naja naja kaouthia (monocellate cobra) venom has been reported (Tan and Swaminathan, 1992), but its structural characterization and physiological function remained to be determined. The function of snake venom LAOs in hemostasis, especially their effect on platelet aggregation, has been controversial. We determined the N-terminal amino acid sequence of the N. n. kaouthia LAO named K-LAO to be DDRRSPLEECFQQNDYEEFLEIAKNGLKKTxNPKHVXxV (38 residues). The protein data base search revealed that the enzyme had high similarities with other snake venom LAOs. Further, platelet aggregation studies revealed that K-LAO functionally did not induce platelet aggregation in a platelet-rich plasma system, but that it inhibited platelet aggregation induced by agonists such as ADP, collagen and ristocetin in a dose-dependent manner. K-LAO diminished platelet aggregation more intensely under low than high shear stress. This inhibitory activity of K-LAO on either ristocetin-induced or shear-induced platelet aggregation was quenched by addition of catalase. These results indicate that K-LAO functions as an inhibitor to platelet aggregation through the formation of hydrogen peroxide. The enzyme may contribute to the development of a severe hematological disorder due to cobra envenomation.     

长白山白眉蝮蛇蛇毒中类凝血酶的分离纯化方法研究

目的:研究长白山白眉蝮蛇蛇毒中类凝血酶的简单分离纯化方法。方法:采用DEAE-Sephadex A-25及Sephadex G-25层析的方法,比较二者对长白山白眉蝮蛇蛇毒中类凝血酶简单分离纯化的效果。结果:从长白山白眉蝮蛇蛇毒中分离出类凝血酶,SDS-PAGE电泳显示为一条带,分子量大约为35.5kDa,达到电泳纯。理化性质研究表明,此类凝血酶具有体外凝血活性,体外凝血酶比活力为12.57IU.mg-1,用N-苯甲酰-L-精氨酸乙酯盐酸盐测得该酶的精氨酸酯酶比活力为137.65IU.mg-1。用蛋白酶抑制剂和乙二胺四乙酸对该酶进行抑制实验,结果表明该酶属于丝氨酸蛋白酶,而不是金属蛋白酶。结论:本方法可用于长白山白眉蝮蛇蛇毒中类凝血酶的分离纯化。     

Characterization of the structure and function of three phospholipases A2 from the venom of Agkistrodon halys pallas

Three monomeric phospholipases A2 with isoelectric points 4.5, 6.9 and 9.3 were purified from the venom of Agkistrodon halys pallas. The complete amino acid sequence of the acidic enzyme and partial amino acid sequences of the neutral and basic phospholipases were determined in order to relate differences in enzymatic reactivities, pharmacologic activities and cytotoxicities to aspects of structure. Studies reported here and elsewhere demonstrate that the three phospholipases A2 exhibit pronounced differences relative to function. The acidic enzyme maintains the highest reactivity toward hydrolysis of monolayers at the air-water interface and may share a feature in common with the acidic enzyme from A. h. blomhoffii, namely the inhibition of platelet aggregation. The neutral phospholipase A2 designated agkistrotoxin, is characterized by potent activity as a pre-synaptic neurotoxin. Agkistrotoxin is the first single polypeptide chain, neurotoxic phospholipase A2 to be documented with a Group II disulfide pattern and, in several respects, may be considered functionally and structurally analogous to notexin from the Australian tiger snake venom. Finally, the basic membranes in the presence of a bactericidal-permeability-increasing protein from neutrophil sources.     

Inhibition of human platelet aggregation by -amino acid oxidase purified from Naja naja kaouthia venom

-Amino acid oxidase (LAO) widely exists in snake venoms. Purification of LAO from the Naja naja kaouthia (monocellate cobra) venom has been reported (Tan and Swaminathan, 1992), but its structural characterization and physiological function remained to be determined. The function of snake venom LAOs in hemostasis, especially their effect on platelet aggregation, has been controversial. We determined the N-terminal amino acid sequence of the N. n. kaouthia LAO named K–LAO to be DDRRSPLEECFQQNDYEEFLEIAKNGLKKTxNPKHVXxV (38 residues). The protein data base search revealed that the enzyme had high similarities with other snake venom LAOs. Further, platelet aggregation studies revealed that K–LAO functionally did not induce platelet aggregation in a platelet-rich plasma system, but that it inhibited platelet aggregation induced by agonists such as ADP, collagen and ristocetin in a dose-dependent manner. K–LAO diminished platelet aggregation more intensely under low than high shear stress. This inhibitory activity of K–LAO on either ristocetin-induced or shear-induced platelet aggregation was quenched by addition of catalase. These results indicate that K–LAO functions as an inhibitor to platelet aggregation through the formation of hydrogen peroxide. The enzyme may contribute to the development of a severe hematological disorder due to cobra envenomation.     

三种蛇毒类凝血酶对纤维蛋白原的作用方式

目的以纤维蛋白原为底物,考察巴西矛头蝮蛇(Bothrops atrox)、尖吻蝮蛇(Agkistrodon acutus)、长白白眉蝮蛇(Agkistrodon halys pullas)3种蛇毒类凝血酶对纤维蛋白原的作用方式。方法采用SDS-PAGE及RP-HPLC法对作用结果进行检测。结果 3种蛇毒类凝血酶对纤维蛋白原的作用方式不完全相同,巴西矛头蝮蛇、尖吻蝮蛇2种蛇毒类凝血酶只作用于纤维蛋白原的α链,对β、γ链则无作用,长白白眉蝮蛇毒类凝血酶起初作用于纤维蛋白原的β链,对α链作用较弱,随着时间的延长对α链作用增强,对γ链无作用。结论巴西矛头蝮蛇、尖吻蝮蛇2种蛇毒类凝血酶属于SVTLE-A型,而长白白眉蝮蛇毒类凝血酶则属于SVTLE-AB型。     

蝮蛇毒纤溶酶的分离纯化及性质研究

目的 :寻找一种分离纯化蝮蛇毒纤溶酶的工艺并研究其理化性质。方法 :采用DEAE SepharoseCL 6B和HeparinCL 6B层析方法 ,从蝮蛇毒中分离纯化纤溶酶。结果 :蝮蛇毒纤溶酶经HPLC为单一峰 ,等电聚焦电泳为一条带 ,其等电点为 4.5 5 ,经SDS 聚丙烯酰胺凝胶电泳测得分子量为 2 9.4kD。该酶对热不稳定 ,在 pH6～ 9时稳定 ,氨基酸组成分析表明含酸性氨基酸较多。结论 :用此工艺可制得高纯度的蝮蛇毒纤溶酶。     

中国黑眼镜蛇毒蛋白酶natrahagin抑制血小板聚集和动脉血栓形成的作用

研究从中国黑眼镜蛇毒中纯化的中国黑眼镜蛇毒蛋白酶natrahagin对血小板聚集 ,血浆纤维蛋白原水平和动脉血栓形成的影响 .采用比浊法测定兔血小板聚集率 ,双缩脲法测定血浆纤维蛋白原浓度 .应用胰蛋白酶损伤血管内皮的方法制作家兔颈动脉血栓模型评价natrahagin抑制血栓形成的作用 .结果发现 ,新西兰家兔natrahagin ( 0 .0 2 5～ 0 .1mg·kg- 1,iv)剂量依赖性地抑制二磷酸腺苷 ( 10 μmol·L- 1)和胶原 ( 10 0mg·L- 1)诱导的血小板聚集 ,显著降低血浆纤维蛋白原水平 .剂量为 0 .1mg·kg- 1时 ,血浆纤维蛋白原水平降低 33.3% .并能有效地抑制兔颈动脉血栓的形成 ,效应呈剂量依赖性 .0 .1mg·kg- 1剂量组对血栓形成的抑制率达到 4 5.4 % .研究提示 ,natrahagin抑制动脉血栓形成的作用与其抑制纤维蛋白原介导的血小板聚集和降低血浆纤维蛋白原水平的作用有关     

Mechanism of action of the platelet aggregation inhibitor purified from Agkistrodon halys (mamushi) snake venom

The platelet aggregation inhibitor purified from Agkistrodon halys snake venom inhibited rabbit platelet aggregations induced by thrombin, sodium arachidonate, collagen or ionophore A-23187. The ic₅₀ was about 11 μg/ml in platelet aggregation regardless of which aggregation inducer was used. β-Mercaptoethanol abolished both the phospholipase A enzymatic and platelet aggregation inhibitory activities of this venom inhibitor. p-Bromophenacyl bromide-treated venom inhibitor lost almost completely its phosphilipase A enzymatic activity, but retained its platelet aggregation inhibitory effect. In the presence of EGTA, the venom inhibitor still showed the same inhibitory activity on thrombin-, sodium arachidonate-, collagen- or ionophore A23187-induced platelet aggregations triggered by successive addition of Ca²⁺. The activation of platelet phospholipase A and the serotonin release reaction triggered by Ca²⁺ influx were unaffected by this venom inhibitor. It also inhibited the clot retraction of platelet-rich plasma. It is concluded that the inhibitory effect of the venom inhibitor on platelet aggregation is independent of its phospholipase A enzymatic activity. Its mode of action is different from those of other known platelet inhibitory drugs. This venom inhibitor possibly acts on a common step subsequent to platelet shape change, leading to inhibition of platelet aggregation.