蛇床子素抑制血栓形成和血小板聚集的实验研究

目的　观察蛇床子素抑制血栓形成和血小板聚集的作用。方法　利用大鼠动-静脉旁路血栓形成模型和小鼠尾静脉注射胶原-肾上腺素合剂诱导血栓形成模型,分别测定给蛇床子素10、20、40mg·kg-1后血栓湿重和干重, 5min内小鼠死亡数和15min内偏瘫恢复数;分别采用ADP、凝血酶、花生四烯酸钠为诱导剂,测定经不同浓度蛇床子素处理后1、3、5min时血小板聚集率及最大聚集率的改变。结果　蛇床子素可以抑制大鼠动-静脉旁路血栓形成,减轻血栓湿重及干重;抑制胶原-肾上腺素合剂诱导的血栓形成,降低5min内的小鼠死亡率,提高15min内偏瘫小鼠的恢复率;蛇床子素在体外可抑制ADP、凝血酶、花生四烯酸钠诱导的血小板聚集,其IC50分别为0 .44、0 .186、0 .421g·L-1。结论　蛇床子素可明显抑制血栓形成和血小板的聚集。     

银杏总内酯抗血小板聚集与抗血栓作用

目的：探讨银杏总内酯抗血栓形成及抑制血小板聚集的作用.方法：采用大鼠动静脉旁路血栓模型和Chandler氏血栓形成法,利用血小板活化因子（platelet activating factor,PAF）、花生四烯酸（arachidonic acid,AA）和二磷酸腺苷（adenosine diphosphate,ADP）诱导家兔血小板聚集,测定血栓形成抑制率、血小板在不同时间点的聚集率以及最大聚集率.结果：银杏总内酯可不同程度地抑制大鼠动静脉旁路血栓和Chandler氏体外血栓形成,减轻血栓重量,血栓形成抑制率分别达到 41.58%和 59.31%;银杏总内酯可抑制PAF和ADP诱导的家兔血小板在不同时间点的聚集,降低其最大聚集率.结论：银杏总内酯可明显对抗血栓形成,并具有显著的抗PAF和ADP诱导血小板聚集作用.     

L—精氨酸L—门冬氨酸盐对血栓形成的影响及其机制

目的观察L-精氨酸L-门冬氨酸盐(DR)对动物血栓形成模型的影响并初步探讨其作用机制.方法用大鼠颈动脉血栓模型、动静脉旁路血栓模型和小鼠肺栓塞模型评价DR的抗血栓作用;测定血浆TXA2、PGI2和NO水平,测定血管内皮释放PGI2水平,以探讨DR的作用机制.结果DR7.5、15、30mg*kg-1单次灌胃给药,可减轻大鼠颈动脉血栓重量(P<0.01);7.5、15、30或60mg*kg-1均可显著抑制血小板在动静脉旁路中丝线上的沉积(P<0.05或P<0.01);但对花生四烯酸引起的小鼠肺栓塞死亡无明显作用.DR30mg*kg-1单次给药(ig),对大鼠血浆TXA2水平无明显影响;使PGI2有升高趋势.而相同剂量阿司匹林(ASA)可明显抑制二者水平.DR30mg*kg-1给药(ig)7次,每日2次,可明显促进血管内皮释放PGI2;并使血浆NO水平有明显升高.结论DR可明显抑制动脉血栓形成,其作用可能与血管内皮释放PGI2和NO有关,而与血小板花生四烯酸代谢途径无关.     

祛栓灵胶囊抑制动物血小板聚集功能的研究

目的观察祛栓灵胶囊对动物血小板聚集功能的影响。方法采用ADP、凝血酶和花生四烯酸三种诱导剂,测定正常大鼠和血瘀模型兔服用祛栓灵胶囊7d后的血小板聚集性变化。结果大鼠服用祛栓灵胶囊62．5～250mg·kg^-1 7d后,对ADP和凝血酶诱导的血小板最大聚集率分别可抑制13．8％～24．0％和11．6％～35．3％;血瘀模型兔服用祛栓灵胶囊37．5～150mg·kg^-1 7d后,对ADP和凝血酶诱导的血小板最大聚集率分别可抑制22．1％～33．9％和16．3％～23．6％。但正常大鼠和血瘀模型兔服用祛栓灵胶囊后,对花生四烯酸诱导的血小板聚集均未见有明显影响。结论祛栓灵胶囊可明显抑制大鼠和兔体内由ADP和凝血酶介导的血小板聚集。     

2-萘-3-(3,4-二甲氧基)苯丙烯酸抗血小板聚集作用及其机理

研究发现,NMPA在体外可抑制花生四烯酸、胶原及凝血酶诱导的家兔血小板聚集,IC_(50)分别为189±9,105±11.0,49.8±16.6μM。小鼠ig NMPA(25mg/kg)后,以ADP诱导的血小板聚集明显受到抑制。大鼠ig NMPA后(52mg/kg一次或10mg/(kg·d)连续7d,出血时间延长、胶原诱导的血小板聚集率下降、丙二醛生成减少、但对动脉壁PGI_2样物质影响较小。说明MNPA的作用机理可能在于改变体内TXA_2/PGI_2比值,而达到抑制血小板聚集的效应。     

复方马其通方抗血栓的作用研究

目的:探究复方马其通方(MQT)的抗血栓作用及其作用机制。方法:采用FeCl₃诱导小鼠动脉血栓模型，评价MQT对血栓形成的影响。采用倒置荧光显微镜观察血小板在固化胶原和纤维蛋白原上的粘附或扩展；采用流式细胞仪检测MQT对二磷酸腺苷(ADP)诱导的血小板颗粒释放和细胞内Ca²⁺浓度的影响；使用血小板聚集仪评价MQT对ADP、凝血酶、胶原、花生四烯酸诱导的血小板聚集的影响；采用免疫印迹及酶联免疫法考察MQT对Akt及cAMP信号的影响；结果:MQT可显著延长血栓形成时间，减少管腔内血栓质量；同时，MQT可降低纤维蛋白原水平，延长凝血酶时间；此外，MQT对纤溶功能及血小板相关指标的表达具有显著影响。MQT可显著抑制血小板在固化胶原上的粘附和影响“由外向内”信号的传递；MQT显著抑制ADP及凝血酶诱导的血小板聚集，并抑制ADP诱导的血小板颗粒释放和Ca²⁺的表达；MQT可抑制Akt磷酸化。结论:MQT具有显著的抗血栓作用，可能与调节血小板Akt信号有关。     

祛栓灵胶囊抑制动物血小板聚集功能的研究

目的 观察祛栓灵胶囊对动物血小板聚集功能的影响。方法 采用ADP、凝血酶和花生四烯酸三种诱导剂,测定正常大鼠和血瘀模型兔服用祛栓灵胶囊7 d后的血小板聚集性变化。结果 大鼠服用祛栓灵胶囊62.5~250 mg·kg^-17 d后,对ADP和凝血酶诱导的血小板最大聚集率分别可抑制13.8%~24.0%和11.6%~35.3%;血瘀模型兔服用祛栓灵胶囊37.5~150 mg·kg^-1 7 d后,对ADP和凝血酶诱导的血小板最大聚集率分别可抑制22.1%~33.9%和16.3%~23.6%。但正常大鼠和血瘀模型兔服用祛栓灵胶囊后,对花生四烯酸诱导的血小板聚集均未见有明显影响。结论 祛栓灵胶囊可明显抑制大鼠和兔体内由ADP和凝血酶介导的血小板聚集。     

香草酸抗血小板聚集作用的体内外研究

目的香草酸是一种酚酸类化合物,具有抗氧化、抗炎等药理作用,其抗血栓作用尚未有报道。本实验室前期通过筛选发现香草酸具有良好的抗血小板聚集作用,因此本研究通过体内外实验对香草酸抗血小板聚集的作用进行系统评价。方法采用花生四烯酸(arachidonic acid,AA)、二磷酸腺苷(adenosine diphosphate,ADP)、凝血酶(thrombin,THR)诱导体内外血小板聚集模型,结合凝血四项检测评价香草酸抗血小板聚集及抗血栓的作用。结果体外实验中,香草酸对ADP和AA诱导的血小板聚集具有显著抑制作用,并剂量依赖性抑制ADP诱导的血小板聚集;体内试验中,香草酸(10、30和100 mg/kg)能够剂量依赖性抑制ADP和AA诱导的血小板聚集;同时,香草酸(100 mg/kg)能显著降低纤维蛋白原、增加凝血酶原时间,对活化部分凝血活酶时间和血浆凝血酶时间无明显影响。结论本研究首次发现香草酸对ADP和AA诱导的血小板聚集具有抑制作用,为研发具有自主知识产权的抗血小板聚集药物提供了实验依据。     

噻氯匹啶对血小板聚集功能的抑制作用研究

观察了噻氯匹啶（TP）对血小板聚集的影响，结果表明在体外可显著抑制ADP、花生四烯酸和凝血酶诱导的大鼠或人血小板聚集，其IC50分别为1．035、1.047和0．092mmol／L，在体内连到给药（TP：50－200mg／kg·d－1）7d可抑制ADP和凝血酶诱导的大鼠血小板聚单，提示TP是一个较强的血小板聚集抑制剂。     

莲心碱对血小板聚集、凝血功能和血栓形成的影响

目的探讨莲心碱对大鼠体内血小板聚集和凝血功能的影响,同时评价其抗血栓作用。方法以二磷酸腺苷(ADP)诱导血小板聚集,采用比浊法观察莲心碱对大鼠体内血小板1、5min聚集率和最大聚集率的影响;通过毛细管法和减尾法分别研究莲心碱对小鼠凝血时间和尾出血时间的影响,同时评价莲心碱对大鼠凝血酶原时间(PT)、活化部分凝血激酶时间(APTT)及凝血酶时间(TT)的影响;采用Chandler法及动静脉旁路模型研究莲心碱对大鼠体外血栓和动静脉旁路血栓形成的影响。结果莲心碱5mg·L-1和10mg·L-1均能明显抑制ADP诱导的大鼠体内血小板1、5min聚集率和最大聚集率;明显延长大鼠PT、APTT和TT;明显延长小鼠凝血时间及尾出血时间;不同程度抑制大鼠体外血栓和动静脉旁路血栓形成,减轻血栓湿重和干重。结论莲心碱可明显对抗血栓形成,并具有对抗血小板聚集和凝血作用。     

Anti-thrombotic effects of higenamine

The anti-platelet and anti-thrombotic effects of higenamine, a benzyltetrahydroisoquinoline alkaloid of the roots of Aconitum japonicum (Ranunculaceae), were investigated. The degree of platelet aggregation was measured with platelet rich plasma (PRP). An acute thrombotic condition was induced in mice by the injection of the mixture of collagen and epinephrine. The thrombus formation was induced inside the arterio-venous shunt tube installed between an abdominal aorta and the renal vein of rats. Higenamine showed inhibitory activities to both human and rat platelet aggregation induced by ADP, collagen and epinephrine. It was more inhibitory to epinephrine induced aggregation (IC(50); 19 and 7.2 microM to human and rat platelets respectively) than ADP- or collagen-induced aggregation. The anti-thrombotic effects of higenamine were also observed in both mouse acute thrombosis model and rat arterio-venous shunt (AV-shunt) models. The oral administration of higenamine (50 or 100 mg/kg) increased the recovery rates from the acute thrombotic challenge in mice and lowered the weight of thrombus formed inside the AV-shunt tube in rats.     

新合成磺酰脲类化合物I_4抗血小板聚集及抗血栓作用

目的研究新合成磺酰脲类化合物I4抗血小板聚集及抗血栓作用。方法测定I4对ADP诱导兔血小板聚集、小鼠凝血时间、大鼠颈动脉旁路血栓、大鼠血栓阻塞时间的作用。结果 I4显著抑制二磷酸腺苷(ADP)诱导的体外兔血小板聚集,延长小鼠凝血时间,降低大鼠颈动脉旁路血栓重量,明显延长电刺激诱导的大鼠颈动脉血栓形成时间。结论 I4具有显著的抗血小板聚集及抗凝血作用。     

Pharmacological properties of YM-57029, a novel platelet glycoprotein IIb/IIIa antagonist

The pharmacological properties of YM-57029 [4-[4-(4-carbamimidoylphenyl)-3-oxopiperazin-1-yl]piperidino]acetic acid monohydrochloride trihydrate), a novel glycoprotein IIb/IIIa antagonist were examined in this study. YM-57029 inhibited fibrinogen binding to purified glycoprotein IIb/IIIa, 1000-fold more potently than the tetrapeptide arginine-glycine-aspartic acid-serine (RGDS). YM-57029 concentration-dependently inhibited ADP-, collagen- and high shear stress-induced platelet aggregation, strongly inhibited ATP release from platelets activated by ADP, and enhanced deaggregation of ADP-induced platelet aggregates. In a pro-aggregatory activity study, RGDS or SC-54701A ((S)-3-[3-[(4-amidinophenyl)carbamoyl]propionamido]-4-pentynoic acid monohydrochloride) caused prominent small aggregate formation. At a higher concentration, RGDS induced medium and large size aggregates, and SC-54701A induced medium aggregates. In contrast, YM-57029 produced only a few small and no larger size aggregates. Ex vivo ADP-induced platelet aggregation and platelet retention to collagen-coated plastic beads were dose-dependently inhibited by YM-57029 after intravenous bolus injection in guinea pigs. YM-57029 produced dose-dependent antithrombotic effects in carotid artery thrombosis and arterio-venous shunt thrombosis models in guinea pigs at 10 and 30 microg/kg, respectively. At these doses, YM-57029 prolonged template bleeding time. These results suggest that YM-57029 is a potent glycoprotein IIb/IIIa antagonist which has less pro-aggregatory effect. It may be a promising antiplatelet agent for thromboembolic diseases, and a good prototype for developing an orally active compound.     

抗人血小板膜糖蛋白Ⅲa单克隆抗体抑制兔血栓形成的实验研究

目的　评价抗人血小板膜糖蛋白Ⅲa单克隆抗体SZ21抑制兔血栓形成的能力。方法不同浓度的SZ21（0、10及20μg／ml）分别加入兔富血小板血浆（PRP）中,进行二磷酸腺苷（ADP）诱导的兔血小板聚集试验;体内注射SZ21（1.5mg／kg）,注射前及注射后5、30及60分钟时制备兔PRP,分别进行聚集试验;用颈动静脉旁路血栓模型研究SZ21对兔血栓形成的作用,20只新西兰兔随机分成4组,体内注射SZ21,剂量为A组0.1mg／kg,B组0.4mg／kg,C组0.75mg／kg,D为对照组（SZ391mg／kg）,然后测定血栓重量。结果　体外20μg／ml的SZ21对兔血小板抑制率为80％;SZ21体内注射60分钟时完全抑制血小板聚集功能;血栓模型分组试验,各组平均栓重为A组31mg,B组21mg,C组20.2mg,D组31mg,B、C组与对照组间有明显统计学差异（P＜0.01）。结论　单克隆抗体SZ21能有效抑制兔动静脉旁路血栓形成。     

Anti-platelet and anti-thrombotic effects of triacetylshikimic acid in rats.

Because shikimic acid is the key intermediate in the shikimate pathway in plants and microorganisms, shikimic acid and its derivatives have been described as herbicides and anti-microbial agents. Triacetylshikimic acid (TSA) is an acetylate derivative of shikimic acid. The possible anti-platelet activity and anti-thrombotic efficacy of TSA were evaluated and its effect on arachidonic acid (AA) metabolism and second messengers including cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) was evaluated. After oral pretreatment with TSA, adenosine diphosphate (ADP)-, collagen-, and AA-induced rat platelet aggregation was inhibited ex vivo in a dose-dependent manner. In an arteriovenous-shunt thrombosis model, oral administration of TSA resulted in a dose-dependent inhibition of thrombus growth. TSA markedly increased the cAMP level and showed no effect on the cGMP level in rat platelets. Also, no significant changes in ADP-induced thromboxane B2 formation in rat platelets or 6-keto-prostaglandin F 1alpha production from the abdominal aorta were observed after oral administration of low and medium doses of TSA (12.5 and 50 mg/kg). Additionally, prothrombin time, activated partial thromboplastin time, and thrombin time were unchanged at effective anti-platelet doses of TSA. These results demonstrate that TSA exerts oral anti-platelet and anti-thrombotic efficacy without perturbation of systemic hemostasis in rats, which was partially concerned with the elevation of cAMP in platelets.     

Antithrombotic effect of a novel tissue plasminogen activator from the venom of Gloydius brevicaudus viper

OBJECTIVE To investigate the thrombolytic and antiplatelet effects of a novel plasminogen activator from the venom of Gloydius brevicaudus viper(GBV-PA)in vitro and in vivo.METHODS Thrombolytic experiments were performed in rabbit models of ear vein thrombosis and carotid artery thrombosis,and in dog model of acute cerebral infarction.Inhibition of thrombus formation was evaluated in rat inferior vena cava thrombosis model and ferric chloride-induced arterial thrombosis.In vitro,we assayed the antithrombotic effect of GBV-PA on rabbit blood clots,euglobulin lysis time(ELT)of rabbit plasma,and on ADP-induced platelet aggregation.RESULTS GBV-PA intravenous administ ration significantly reduced vascular recanalization times of rabbit ear veins thrombosis and thrombus weight of rabbit carotid artery thrombosis.The arterial recanalization rates were dose-and time-dependently improved after administration of GBV-PA in canine acute cerebral infarction model.Thrombus length and weight was significantly reduced by GBV-PA both in rat inferior vena cava and ferric chloride-induced arterial thrombosis models.Thrombus formation of blood of rabbits they were administrated with GBV-PA was also inhibited.GBV-PA radically reduced plasma ELT of rabbit′s blood clots.ADP-induced platelet aggregation was inhibited by GBV-PA in a dose-dependent manner with a half-maximal inhibitory concentration of 19.9μg·mL-1.CONCLUSION This study demonstrates that GBV-PA is a thrombolytic and antiplatelet agent.It has significant antithrombotic effects on various in vitro and in vivo experimental models of thrombosis.The mechanisms that underline its antithrombotic effects were related to GBV-PA′s capabilities of increasing fibrinolytic activities and inhibition of platelet aggregation.     

国产注射用比伐卢定对大鼠血栓模型血小板活化功能的影响

目的探讨国产注射用比伐卢定（domesticbivalimdin）对大鼠颈总动脉血栓模型血小板活化功能的影响。方法建立大鼠左颈总动脉血栓模型,分为对照组（n=6）、模型组（n=10）、国产注射用比伐卢定联合尿激酶（urokinase,UK）组（B＆UK组,n=10）、肝素（heparin）联合尿激酶组（H＆UK组,n=10）,检测各组治疗前后GMP-140含量及血栓重量。结果与模型组比较,B＆UK组及H＆UK组血栓重量均明显减轻,而B＆UK组与H＆UK组之间血栓重量比较差异无统计学意义。模型组、B＆UK组及H＆UK组GMP-140含量均显著高于对照组;而治疗后GMP-140含量在B＆UK组显著低于模型组,相反治疗后GMP-140含量在H＆UK组则显著高于模型组。结论国产注射用比伐卢定能在模型中有效抑制血栓形成,但相比肝素,国产注射用比伐卢定能显著抑制血小板活化功能,抑制血小板聚集。在抑制血栓形成过程中国产注射用比伐卢定对血小板活化的影响具有更多优势。     

阿斯匹林与维拉帕米并用对血小板聚集和血栓形成的影响

ASA与Ver对ADP诱导的兔体外血小板聚集和大鼠半体内血小板聚集均有明显的抑制作用,且呈量效关系。两者合用作用增强。ASA与Ver均能明显延长电刺激大鼠颈动脉血栓的形成时间,减少胶原和肾上腺素复合诱导剂静脉注射所引起的小鼠肺血栓形成的死亡。剂量与效应相关。两者合用作用增强。     

Antithrombotic and thrombolytic activities of Agkisacutacin, a snake venom proteinase, in experimental models

The antithrombotic and thrombolytic activities of Agkisacutacin (Agk), a component isolated from Agkistrodon acutus, were determined in vitro and in vivo. The models employed included Chandler's model, arterio-venous shunt model and pulmonary embolus model. The effects of Agkisacutacin on coagulation, plasma fibrinogen and platelet aggregation induced by collagen, adenosine diphosphate (ADP) and thrombin were also investigated. The results showed that Agkisacutacin can significantly inhibit thrombus formation in Chandler's and arterio-venous shunt models, and accelerate thrombolysis of pulmonary emboli in rats. The data suggested that Agkisacutacin possessed antithrombotic and thrombolytic activities. Agkisacutacin was also partial characterized.