冠心病患者纤维激活系统功能改变的初步观察

对１８例不同类型的冠心病患者及６例正常人的血浆组织型纤溶酶原激活物及其抑制物的活性进行检测，结果表明，心肌梗塞后及不稳定性心绞痛患者ＰＡＩ－１活性显着增高，ｔＰＡ活性显著下降，ＰＡＩ－１／ｔＰＡ活性比值显着增高；稳定性心绞痛患者ＰＡＩ－１活性轻度增高，ｔＰＡ活性及ＰＡＩ１／ｔＰＡＩ活性比值变化不明显，全部受检者ＰＡＩ－１与ｔＰＡ活性具有显著线性负相关，提示冠心病纤溶激活功能减低，该损害可能具有冠心     

极化液对冠心病患者纤溶系统功能的影响

目的：观察极化液葡萄糖－胰岛素－氯化钾（ＧＩＫ）用于冠心病治疗时对纤溶系统功能的影响。方法：４６例确诊冠心病患者,其中陈旧性心肌硬塞１０例,不稳定性心绞痛１４例,稳定性心绞痛２２例。给予ＧＩＫ液,即１０％葡萄糖５００ｍｌ＋普通胰岛素１０μｌ＋氯化钾１ｇ静滴 ８～１２ ｄ。于用药前后测定组织型纤溶酶原激活物活性（ｔ－ＰＡ：Ａ）及其抑制物活性（ＰＡＬ：Ａ）,纤溶酶原活性（ＰＬｇ）,组织型纤溶酶原激活物（ｔ－ＰＡ）及其抑制物（ＰＡＩ－Ｉ）含量及Ｄ－二聚体（Ｄ－Ｄｉｍｅｒ）含量,空腹血清胰岛素水平。结果：用药前与正常比较,ｔ－ＰＡ活性降低,ＰＡＩ－１活性增高,胰岛素水平增高,高胰岛素者占７８．２％。用药后ｔ－ＰＡ活性及含量、纤溶酶原活性均较前降低（Ｐ＜０．０１）,ＰＡＩ－１含量较前增加（Ｐ＜０．００１）,而ＰＡＩ－１活性及Ｄ－Ｄｉｍｅｒ变化不明显。结论：冠心病特别伴有胰岛素水平增高患者,应用ＧＩＫ液可能通过其对纤溶系统功能的不良影响促使病变进展。     

参三七皂甙Rg1对实验性血栓形成的影响及其机制探讨

用大鼠动静脉血栓形成模型，研究参三七皂甙Ｒｇ１抗血栓作用。结果表明，参三七皂甙Ｒｇ１可明显降低实验性血栓形成，对大鼠血浆纤溶系统亦有明显作用，可升高血浆中组织纤溶酶原激活物（ｔＰＡ）活性和活性型ｔＰＡ百分比，降低组织纤溶酶原激活物抑制剂（ＰＡＩ）活性。同时利用培养大鼠血管内皮细胞实验，发现Ｒｇ１可以剂量依赖性提高血管内皮细胞一氧化氮（ＮＯ）释放。提示Ｒｇ１抗血栓作用与增强纤溶系统活性，促进血管内皮ＮＯ释放有关     

冠心病病人血浆纤溶成分与血脂水平评价

探讨冠心病（ＣＨＤ）患者血浆组织型纤溶酶原激活剂（ｔ－ＰＡ）、纤溶酶原激活剂抑制剂（ＰＡＩ）、交联纤维蛋白二聚体（Ｄ－ｄｉｍｅｒ）及脂蛋白（ａ）［ Ｌｐ（ａ）］等血浆水平变化、临床应用价值以及其相互间的关系。方法： ｔ－ＰＡ、ＰＡＩ、ＬＰ［ａ］、Ｄ－ｄｉｍｅｒ采用ＥＬＩＳＡ双抗体夹心法检测,胆固醇、甘油三酯采用酶法测定。结果：４７例ＣＨＤ ,患者血浆ｔ－ＰＡ－ＰＡＩ及Ｄ－ｄｉｍｅｒ测值为（９．７±５．７）、（８０．５±３０．０）、（２０９±１２５）ｎｇ ／ｍｌ,其中ｔ－ＰＡ,和Ｄ－ｄｉｍｅｒ值明显高于健康对照组（Ｐ＜０．００１）,ＰＡＩ含量两组水平差别不明显（Ｐ＞０．０５）．血浆ＬＰ（ａ）、ＴＧ及ＨＤＬ浓度两组间差别有统计学意义（Ｐ＜０．０５）。用准确性评价各项指标临床应用价值,以Ｄ－ｄｉｍｅｒ为最佳,Ｌｐ（ａ）次之。血浆ｔ－ＰＡ与Ｌｐ（ａ）及ＴＧ含量呈正相关。ＰＡＩ与血浆ＴＧ呈正相关,和ＨＤＬ呈负相关。结论：ｔ－ＰＡ、Ｄ－ｄｉｍｅｒ、ＬＰ（ａ）与粥样斑块及血栓形成有密切关系。     

脑血栓形成患者血液学变化的探讨

目的 探讨脑血栓形成患者的血液学变化（血粘度、纤溶活性、内皮细胞功能、血浆纤维蛋白原）及两者的相关性。方法 检测４８例脑血栓形成患者急性的血粘度、血浆ｔ－ＰＡ主ＰＡＩ活性、血浆Ｄ－二聚体、血浆ＶＷＦ、血浆Ｆｇ,并与４８例健康人对照分析。结果脑血栓形成患者的血粘度,血浆ＰＡＩ活性、Ｄ－二聚体、ＶＷＦ、Ｆｇ均明显高于对照组;血浆ｔ－ＰＡ活性低于对照组。结论由于血管壁受到损伤,内皮细胞的促栓功能加强,     

填精补血化瘀口服液对实验性动脉硬化鹌鹑纤溶活性的影响及机理研究

填精补血化瘀口服液具有升高食铒性动脉硬化（Ａｔｈｅｒｏｓｃｌｅｒｏｓｉｓ，ＡＳ）鹌鹑血浆组织型纤溶酶元激活物（Ｔｉｓｓｕｅｐｌａｓｍｉｎｏｇｅｎａｃｔｉｖｉｔｏｒ，ｔ－ＰＡ）活性，降低纤溶酶元激活物抑制物（Ｐｌａｓｍｉｎｏｇｅｎａｃｔｉｖｉｔｏｒｉｎｈｉｂｉｔｏｒ，ＰＡＩ）活性；调整二者活性比，恢复纤维系统功能，其作用机理可能与该方药减少动脉壁脂质沉积，改善血液流变性，减轻动脉内膜损伤，保护平滑肌     

普伐他汀的抗凝和纤溶功能对冠心病的影响

目的：观察普伐他汀治疗冠心病病人时的抗凝和纤溶功能。方法：２０例冠心病病人［其中５例总胆固醇（ＴＣ）升高，６例三酰甘油（ＴＧ）升高，２例ＴＣ，ＴＧ均升高］，用普伐他汀１０ｍｇ，ｐｏ，ｑｎ，连服４ｗｋ。结果：治疗后组织型纤维蛋白溶酶原激活物（ＴＰＡ）活性明显升高（升高０．２±０．３ＩＵ／ｍＬ），Ｐ＜０．０１，ＴＰＡ与纤维蛋白溶酶原激活剂抑制物（ＰＡＩ）的比值（ＴＰＡ／ＰＡＩ）升高０．３±０．６（Ｐ＜０．０５）。抗凝血酶ＩＩ（Ａｔ－ＩＩ）活性、Ａｔ－ＩＩ抗原、ＰＡＩ、凝血因子Ｉ及Ｄ－二聚体无显著性改变。而对其中ＴＣ和ＴＧ正常者治疗后降脂疗效不明显，Ｐ＞０．０５。结论：普伐他汀有促进纤溶活性作用。     

慢性肺心病急性发作期t—PA,PAI活性的变化

测定３２例肺心病急性发作期组织型纤溶酶原激活物（ｔ－ＰＡ）、纤溶酶原激活物抑制物（ＰＡＩ）、纤溶酶原（Ｐｇ）及纤维蛋白原降解产物（ＦＤＰ）。结果表明，肺心病患者ｔ－ＰＡ、ＰＡＩ、Ｐｇ明显下降，而ＦＤＰ明显升高，表明肺心病急性发作期处于高凝、低纤溶状态。ｔ－ＰＡ、Ｐｇ的变化与ＰａＯ２呈明显正相关，而ＦＤＰ与ＰａＯ２呈明显负相关。     

填精补血化瘀口服液对实验性动脉硬化鹌鹑纤溶活性的影响及机理研究

填精补血化瘀口服液具有升高食饵性动脉硬化（Ａｔｈｅｒｏｓｃｌｅｒｏｓｉｓ，ＡＳ）鹌鹑血浆组织型纤溶酶元激活物（ＴｉｓｓｕｅＰｌａｓｍｉｎｏｇｅｎａｃｔｉｖｉｔｏｒ，ｔ－ＰＡ）活性，降低纤溶酶元激活物抑制物（Ｐｌａｓｍｉｎｏｇｅｎａｃｔｉｖｉｔｏｒｉｎｈｉｂｉｔｏｒ，ＰＡＩ）活性；调整二者活性比，恢复纤溶系统功能。其作用机理可能与该方药减少动脉壁脂质沉积，改善血液流变性，减轻动脉内膜损伤，保护平滑肌细胞及内皮细胞功能活动有关．因而，该方药对治疗ＡＳ，尤其是防止ＡＳ的恶化，都具有积极意义。     

产组织型纤溶酶原激活剂CHO工程细胞无血清培基的研究

在对ＤＭＥＭ：Ｆ１２（１：１）进行优化的基础上,以细胞生长和组织型纤溶酶原激活剂（ｔ－ＰＡ）表达为评价指标,筛选无血清培养基添加成分,建立了由优化ＤＭＥＭ：Ｆ１２（１：１）,胰岛素,ＰｌｕｒｏｎｉｃＦ－６８维生素Ｃ,乙醇胺和微量元素混合液组成的适于产ｔ－ＰＡＣＨＯ工程细胞４Ｂ３生长和ｔ－ＰＡ表达效果均优于相同条件下用含血清培基培养的效果。     

Inhibition of PAI-1: a new anti-thrombotic approach

Proteolytic degradation of fibrin (fibrinolysis) is mediated by plasminogen and its activators, tissue-type plasminogen activator (tPA(1)) and urokinase (uPA). Fibrinolysis is critical for preventing thrombus growth and restoring blood flow following thrombotic vascular occlusion. Plasminogen activator inhibitor-1 (PAI-1), a member of the serine protease inhibitor (serpin) superfamily, is the principal inhibitor of tPA and uPA in the fibrinolytic system. High levels of circulating PAI-1 are associated with a number of thrombotic diseases. In animal studies, transgenic mice overexpressing human PAI-1 develop spontaneous thrombosis, whereas PAI-1-deficient mice are more resistant to venous or arterial thrombosis. Furthermore, inhibition of PAI-1 activity prevents thrombus formation in animal models. The antithrombotic effects of PAI-1 inhibition are achieved by enhancing endogenous fibrinolytic activity without directly affecting blood coagulation and platelet function. Phenotypic analysis of PAI-1 deficiency in both human and mouse suggests that inhibition of PAI-1 will not lead to severe bleeding or other major adverse effects. Thus, PAI-1 inhibitors represent a new class of antithrombotic drugs with a possible wider therapeutic index than conventional antiplatelet and anticoagulant agents. This review summarizes the role of PAI-1 in thrombotic diseases and recent progress in the development of small molecule PAI-1 inhibitors.     

过敏性紫癜肾儿童血尿纤溶酶原激活物及其抑制物的变化及干预治疗

目的　探讨过敏性紫癜肾患儿外周血和尿液中组织型纤溶酶原激活物 (tPA)及 1型纤溶酶原激活物抑制物 (PAI 1)的活性和尿纤溶酶原激活物总活性 (PAs)的变化特点及予血管紧张素转化酶抑制剂 (ACEI)、低分子肝素治疗的影响。方法　 2 3例紫癜肾儿童为研究对象 ,11名正常对照。分别于ACEI和低分子肝素治疗前、治疗后 1周、治疗后 2周取血、尿标本 ,用纤维平板法测定尿PAs活性 ,用酶联免疫吸附测定 (ELISA)法测定血tPA和PAI 1活性 ,以 x±s报道数据。 结果　紫癜肾患儿于治疗前血tPA水平和尿PAs总活性均明显低于正常对照组 ,而PAI 1水平则显著高于对照组 ,ACEI和肝素联合治疗后 ,随着治疗的持续可明显改善凝血纤溶系统平衡紊乱的趋势 (P <0 0 1)。结论　紫癜肾儿童血、尿凝血纤溶系统平衡紊乱 ,表现为凝血亢进和纤溶障碍 ,干预性治疗具有一定疗效     

Modulation of PAI-1 and tPA activity and thrombolytic effects of corilagin

In this study, Charlton's and Tomihisa's methods were modified to investigate the thrombolytic effect of corilagin from the Chinese herbal plant Phyllanthus urinaria L., as well as its effect on carotid artery patency status. The activity of type 1 plasminogen activator inhibitor (PAI-1) in rat plasma or platelet-released substances and tissue-type plasminogen activator (tPA) in rat plasma was assayed by use of a chromogenic substrate. The results showed that corilagin had a dose-dependent thrombolytic effect in rats. 5 mg/kg of corilagin produced a nearly similar reperfusion rate to that of 20000 U/kg of urokinase, whereas it produced a lower reocclusion rate than urokinase. Corilagin significantly inhibited PAI-1 activity in rat plasma or platelet-released substances while it elevated plasma tPA activity, in a concentration-dependent manner. Corilagin, however, had no influence on rabbit platelet aggregation. It is indicated that corilagin inhibited PAI-1 activity and increased tPA activity, and this property of corilagin is assumed to be responsible for the thrombolytic effect. Abbreviations. PO:persistent occlusion CR:cyclic reflow PP:persistent patency PAI-1:type 1 plasminogen activator inhibitor tPA:tissue-type plasminogen activator PBS:phosphate buffer solution IC (50):50 % of inhibitory concentration PRP:platelet-rich plasma ADP:adenosine diphosphate AA:arachidonic acid PAF:platelet-activating factor     

扎冲十三味丸的溶栓作用机制及对凝血系统的影响

目的:探讨扎冲十三味丸的溶栓作用及其初步机制,同时评价其对凝血系统的影响及急性毒理试验。方法:采用体内和体外溶栓的方法评价扎冲十三味丸对电刺激大鼠颈动脉血栓的溶栓作用;应用ELISA方法测定扎冲十三味丸对家兔血浆组织型纤溶酶原激活物(tPA)和纤溶酶原激活物抑制物(PAI-1)活性的影响;同时评价扎冲十三味丸凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、凝血酶时间(TT)、纤维蛋白原(Fib)含量和血液流变学的影响。扎冲十三味丸对实验小鼠的急性毒理试验。结果:0.432 g· kg^-1(中)和0.864 g· kg^-1(高)扎冲十三味丸组于末次给药后2~6 h均明显抑制血浆PAI-1活性, 显著提高血浆tPA活性。0.864,1.728 g·kg^-1组和复方丹参滴丸0.12 g·kg^-1组明显降低Fib水平,明显延长TT,3种剂量的扎冲十三味丸对APTT和PT无明显影响,对红细胞电泳、高切、中切和低切均无明显影响;全部小鼠健存,无中毒反应。结论:扎冲十三味丸抑制PAI-1活性,同时提高tPA的活性可能是其具有较好溶栓作用的分子机制。扎冲十三味丸可影响内凝和外凝系统,减少血管壁纤维蛋白的沉积,也提示在临床用药过程中,有可能发生出血倾向,需密切监测患者的凝血系统;无明显急性毒理反应。     

Losartan inhibits the angiotensin II-induced modifications on fibrinolysis and matrix deposition by primary human vascular smooth muscle cells

Disorders in the fibrinolytic and renin-angiotensin-aldosterone systems and excessive extracellular matrix (ECM) deposition are determinant factors in several pathologic manifestations of vascular and cardiac tissue. We used primary human vascular smooth muscle cells (VSMC) and studied the effects of losartan on angiotensin II (Ang II)-mediated (a) DNA synthesis, (b) secretion of tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1), (c) secretion of matrix metalloprotease-2 (MMP-2) activity and tissue inhibitors of MMPs (TIMPs), and (d) synthesis of glycosaminoglycans. VSMC cultures, established from human pulmonary arteries, were treated with Ang II (0.1 nM -1 microM ) and/or losartan (0.1-10 microM ), for 24 or 48 h. DNA synthesis was assessed by incorporation of 3 H-thymidine into VSMC, secreted tPA, PAI-1, and TIMPs by enzyme-linked immunosorbent assay, MMP-2 activity by gelatin zymography, and glycosaminoglycan synthesis by 3 H-glucosamine incorporation. Ang II (1 microM ) enhanced DNA synthesis and secretion of PAI-1 and glycosaminoglycans and decreased secretion of MMP-2 activity and tPA, whereas it had no effect on secretion of TIMPs and glycosaminoglycans associated with cell layers. The Ang II-mediated effects were reversed by losartan, in a concentration-dependent manner. Losartan alone increased secretion of tPA, MMP-2 activity, and TIMPs and decreased secretion of PAI-1. These results indicate that AT 1 receptors are implicated in Ang II-mediated disorders of fibrinolysis and excessive ECM deposition by VSMC.     

Neutralization of plasminogen activator inhibitor I (PAI-1) by the synthetic antagonist PAI-749 via a dual mechanism of action

PAI-749 is a potent and selective synthetic antagonist of plasminogen activator inhibitor 1 (PAI-1) that preserved tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA) activities in the presence of PAI-1 (IC(50) values, 157 and 87 nM, respectively). The fluorescence (Fl) of fluorophore-tagged PAI-1 (PAI-NBD119) was quenched by PAI-749; the apparent K(d) (254 nM) was similar to the IC(50) (140 nM) for PAI-NBD119 inactivation. PAI-749 analogs displayed the same potency rank order for neutralizing PAI-1 activity and perturbing PAI-NBD119 Fl; hence, binding of PAI-749 to PAI-1 and inactivation of PAI-1 activity are tightly linked. Exposure of PAI-1 to PAI-749 for 5 min (sufficient for full inactivation) followed by PAI-749 sequestration with Tween 80 micelles yielded active PAI-1; thus, PAI-749 did not irreversibly inactivate PAI-1, a known metastable protein. Treatment of PAI-1 with a PAI-749 homolog (producing less assay interference) blocked the ability of PAI-1 to displace p-aminobenzamidine from the uPA active site. Consistent with this observation, PAI-749 abolished formation of the SDS-stable tPA/PAI-1 complex. PAI-749-mediated neutralization of PAI-1 was associated with induction of PAI-1 polymerization as assessed by native gel electrophoresis. PAI-749 did not turn PAI-1 into a substrate for tPA; however, PAI-749 promoted plasmin-mediated degradation of PAI-1. In conclusion, PAI-1 inactivation by PAI-749 using purified components can result from a dual mechanism of action. First, PAI-749 binds directly to PAI-1, blocks PAI-1 from accessing the active site of tPA, and abrogates formation of the SDS-stable tPA/PAI-1 complex. Second, binding of PAI-749 to PAI-1 renders PAI-1 vulnerable to plasmin-mediated proteolytic degradation.     

福辛普利对慢性肾功能衰竭幼鼠凝血纤溶变化的干预影响

目的探讨血管紧张素转换酶抑制剂(ACEI)——福辛普利干预治疗对幼年大鼠慢性进展性肾脏病变过程中肾组织组织型纤溶酶原激活物(tPA)、组织型纤溶酶原激活物特异性抑制剂(PAI)-1蛋白表达的影响。方法以5/6肾切除SD大鼠为实验动物模型,予ACEI治疗12周后分别检测大鼠体重、血压、血尿生化及残肾组织常规病理和用免疫组织化学染色定性定量评价残肾tPA、PAI-1蛋白表达。结果于5/6肾切除后,大鼠肾功能进行性丧失,表现为尿蛋白、血压、血尿素氮(BUN)、肌酐(Scr)及血脂增高,残肾组织出现肾小球硬化和间质纤维化。免疫组织化学染色提示肾组织PAI-1表达增高,而tPA表达下降。经ACEI治疗后,大鼠肾功能改善、血压下降、尿蛋白下降、残肾组织tPA蛋白表达增加、PAI-1表达下降。结论ACEI具有改善5/6肾切除大鼠凝血纤溶系统紊乱的作用。     

Effects of polysulfated glycosaminoglycan and triamcinolone acetonid on the production of proteinases and their inhibitors by IL-1alpha treated articular chondrocytes

In this study we determined the in vitro effects of polysulfated glycosaminoglycan (PSGAG) and the glucocorticoid triamcinolone acetonid (TA) on the IL-1 altered expression and activity of matrix metalloproteinases (MMP-1, MMP-3), tissue inhibitor of metalloproteinases-1, the plasminogen activators tPA and uPA and plasminogen activator inhibitor 1 by articular chondrocytes. Bovine chondrocytes were cultured in alginate gel beads. Cells were treated with interleukin-1alpha (IL-1alpha) in the presence of vehicle or drugs at various concentrations. After 48hr mRNA expression of MMP-1, MMP-3, TIMP-1, uPA, tPA and PAI-1 was analyzed by RT-PCR-ELISA. The protein synthesis of TIMP-1 and MMP-3 was determined by immunoprecipitation, PAI-1 protein was quantitated by ELISA. The activity of enzymes and inhibitors was measured by functional assays. Treating chondrocytes with IL-1 induced the expression of MMPs and downregulated TIMP-1 but stimulated both the expression of PAs and PAI-1. Both drugs significantly reduced collagenase and proteoglycanase activities which was accompanied by inhibition of the expression of MMP-1 and MMP-3. The IL-1 decreased expression of TIMP-1 was further reduced by TA, which resulted in a significant loss of TIMP activity. No effects on TIMP activity or TIMP-1 biosynthesis were observed after treatment of chondrocytes with PSGAG. Both drugs inhibited the IL-1-induced mRNA expression of tPA, whereas expression of uPA was only mildly reduced by PSGAG, which also induced PAI-1 above IL-1 stimulated levels. As inhibition of collagenase activities and tPA expression by PSGAG occurred at physiological concentrations it might be of clinical relevance, indicating that PSGAG could help reducing cartilage degradation and has a strong anti-fibrinolytic potential. Due to their co-regulation of MMPs and TIMP(s) glucocorticoids should be carefully studied for their overall effect on extracellular matrix proteolysis.     

Novel situations of endothelial injury in stroke--mechanisms of stroke and strategy of drug development: novel mechanism of the expression and amplification of cell surface-associated fibrinolytic activity demonstrated by real-time imaging analysis

Vascular endothelial cells (VECs) secrete tissue plasminogen activator (tPA) in an active form and thus its facilitated secretion directly enhances fibrinolytic activity. We have recently demonstrated its unique secretory dynamics in GFP-tagged tPA expressing VECs using total internal reflection fluorescence microscopy. tPA-GFP appeared to remain on the cell surface after secretion. Studies using a domain-deleted mutant of tPA-GFP suggested that its binding to the cell surface was heavy-chain dependent. PA inhibitor-1 (PAI-1) facilitated dissociation of tPA-GFP by forming a high molecular weight complex. Lack of dissociation from the cell surface of catalytically inactive mutant tPA-GFP, which does not complex with PAI-1, supported PAI-1 dependence of the disappearance of tPA from the VEC surface. To confirm the possibility that retained active tPA modified cell surface fibrinolytic activity, we analyzed binding of Alexa Fluor 568-labeled plasminogen (plg-568) in tPA-GFP expressing cells. Plg-568 appeared to accumulate at tPA-GFP-retained spots as well as in pericellular/matrix adhesive areas. Either modification of the active site or deletion of the tPA-GFP heavy chain resulted in decreased accumulation of plg-568. Prolonged retention appeared essential for tPA to effectively express and amplify fibrinolytic activity on VECs, which may also be responsible for development of deleterious effects in the case of stroke.