人组织因子途径抑制物基因在兔颈动脉损伤血栓形成中的作用

目的 观察人组织因子途径抑制物(tissue factor pathway inhibitor,TFPI)基因在兔颁动脉损伤血栓形成的作用.方法 大耳白兔40只,体质量2.5～3.0 kg.将白兔按体质量随机分为4组:腺病毒介导的TFPI基因(Ad-TFPI)转染组、腺病毒介导的LacZ基因(Ad-LzcZ)转染组、磷酸盐缓冲液(PBS)组和对照组,每组10只.对照组不进行任何处理,另3组用PTCA球囊导管对白兔右侧颈动脉进行损伤,损伤处分别用DisDatch转基因导管按组别局部注射转染Ad-TFPI、Ad-LacZ液和PBS各0.2 ml.转染10 d后,采用相同方法对白兔颈动脉损伤处进行二次损伤,对照组进行第1次损伤,损伤后立即恢复血流.30 min后处死白兔,分别取出损伤侧和对侧颈总动脉,沿长轴剪开、展平,2%戊二醛固定,扫描电镜下观察4组白兔损伤处动脉管壁血小板聚集及纤维素(血栓)形成情况.结果 扫描电镜下,正常未经任何损伤的颈动脉管腔内可见血管内皮细胞结构完整,排列整齐;对照组在球囊损伤后血管内皮不完整,有部分血小板聚集,血管肇上无纤维素生成;Ad-TFPI组在二次损伤后血小板聚集增加,但血管壁上未见纤维素形成;Ad-LacZ组和PBS组可见皿管壁上有大量的纤维素形成和红细胞堆积.血管壁血栓形成阳性率组间比较差异有统计学意义(χ2=14.95,P<0.01);其中Ad-TFPI组(20%)低于Ad-LacZ组(80%,χ2=7.20,P<0.01)和PBS组(70%,χ2=5.05,P<0.05),高于对照组(10%,χ2=0.39,P>0.05);Ad-LacZ组(80%)高于对照组(10%,χ2=9.90,P<0.01),也高于PBS组(70%,χ2=0.27,P>0.05);PBS组(70%,)高于对照组(10%,χ2=7.50,P<0.01).结论 二次球囊损伤法可造成大量血栓形成;腺病毒表达载体TFPI可以抑制动脉损伤后纤维素沉积.     

组织因子途径抑制物基因转移对动脉血栓形成的抑制作用

目的:研究日本凝血病毒(HVJ)-人工病毒包膜(AVE)脂质体介导的组织因子途径抑制物(TFPI)基因转移对损伤血管局部血栓形成抑制作用的有效性及安全性。方法:在兔球囊损伤髂动脉局部分别转染含TFPI基因(TFPI组)或pcDNA3．1质粒(空质粒组)的HVJ-AVE脂质体复合物或HVJ-脂质体(空载体组)或生理盐水(盐水组),1,3和7天后用逆转录多聚酶链反应和免疫组化法检测血管局部TFPI表达;组织病理学和扫描电镜观察比较各组血栓形成情况;各组实验动物观察6个月评价这种疗法的安全性。结果:TFPI基因转移后1天即检测到TFPI mRNA表达,第3天为高峰;TFPI组血栓形成例数明显少于其它3组(P<0.05);体内凝血指标及生化指标无明显变化;重要脏器组织病理学检查未见明显变化;各器官未见到HVJ感染及复制迹象。结论:HVJ-AVE脂质体介导TFPI基因转移能够安全而有效地抑制损伤血管局部血栓形成。     

组织因子在血栓形成中的作用

组织因子（ＴＦ）是一种跨膜糖蛋白,为凝血因子Ⅶａ在细胞表面的受体和辅因子,晃凝血过程的启动因子。生理情况下,与血液直接接触的细胞并不表达ＴＦ。最近的研究表明,ＴＦ的活动与动脉继样硬化所致的血栓形成有密切关系。文章重点探讨ＴＦ在动脉粥样硬化、血栓形成和溶栓后血栓性再闭塞中的作用及其研究进展。     

组织因子在血栓形成中的作用

组织因子（ Ｔ Ｆ） 是一种跨膜糖蛋白，为凝血因子Ⅶａ 在细胞表面的受体和辅因子，是凝血过程的启动因子。生理情况下，与血液直接接触的细胞并不表达 Ｔ Ｆ。最近的研究表明， Ｔ Ｆ 的活动与动脉粥样硬化所致的血栓形成有密切关系。文章重点探讨 Ｔ Ｆ 在动脉粥样硬化、血栓形成和溶栓后血栓性再闭塞中的作用及其研究进展。     

组织因子途径抑制物在动脉粥样硬化中的作用

组织因子与动脉粥样硬化的发生发展密切相关,而组织因子途径抑制物(TFPI)是调节组织因子介导的凝血途径的主要抑制物。大量的实验证据提示,组织因子途径抑制物在一些方面具有抑制作用,包括内皮细胞的活性、巨噬细胞的功能以及平滑肌细胞的增殖和迁移等。本文重点阐述了组织因子途径抑制物对动脉粥样硬化发生发展的抑制作用。     

组织因子途径抑制物与缺血性脑血管病

凝血系统在缺血性脑血管病的发生发展过程中发挥着重要作用。组织因子途径抑制物作为一 种生理性抗凝物质,即外源性凝血途径起始步骤的生理性抑制物,与缺血性脑血管病的关系密切。文 章重点介绍了组织因子途径抑制物在缺血性脑血管病中的变化、作用机制及其临床治疗前景。     

腺病毒介导组织因子途径抑制物基因转染兔损伤颈动脉后的表达及对内膜增生的抑制作用

目的 检测人组织因子途径抑制物(tissue factor pathway inhibitor,TFPI)基因腺病毒载体系统在体表达及对兔颈动脉球囊损伤后新生内膜增生的抑制作用.方法 共采用中国大耳白兔70只.分为4组,分别为腺病毒介导TFPI基因(Ad-TFPI)转染组、腺病毒介导LacZ基因(Ad-LacZ)转染组、PBS组及假手术组,前3组分别于颈动脉球囊损伤后局部转染Ad-TFPI、Ad-LacZ或PBS,假手术组只分离颈总动脉,不做球囊损伤.Ad-TFPI组和Ad-LacZ组每组25只动物,PBS组及假手术组每组10只.Ad-TFPI组和Ad-LacZ组分别于术后3、7、10、14和28 d各处死3只动物,用RT-PCR、ELISA方法 ,检测TFPI在颈动脉壁中的表达.术后4周时,4组剩余动物(每组10只)行颈动脉造影检测最小管腔直径,之后取血管标本,观察病理形态学变化,测算出血管新生内膜面积、内膜与中膜的比例(I/ M)、管腔狭窄程度.结果 基因转染后3 d,Ad-TFPI 组检测到TFPI mRNA及蛋白表达,10～14 d为表达高峰,28 d有所下降但仍可检测到,Ad-LacZ组未测到人TFPI的表达.基因转染后4周,颈动脉造影结果 示Ad-TFPI组最小管腔直径明显大于PBS组和Ad-LacZ组[(1.17±0.43)mm比(0.43±0.33)mm、(0.39±0.24)mm],管腔狭窄百分率明显低于PBS组和Ad-LacZ组[(32±8.2)%比(79±10.2)%、(81±13.1)%],P均<0.01.形态学检查分析结果 显示Ad-TFPI组新生内膜面积明显小于PBS组和LacZ组[(0.17±0.03)mm2比(0.73±0.03)mm2、(0.78±0.04)mm2],L/ M值明显小于PBS组和LacZ组(1.02±0.25比2.76±0.33、2.92±0.24),管腔狭窄程度明显低于PBS组和LacZ组[(24.5±2.1)%比(78.2±2.8)%、(81.3±3.2)%],P均<0.01.结论 人TFPI腺病毒表达载体可在体内有效表达,并且对血管成形术后再狭窄有抑制作用.     

组织因子途径抑制物在心血管疾病中的研究现状

组织因子途径抑制物(tissue factor pathway inhibitor,TFPI)是一种在生理条件下血液中天然存在的抗凝物质,通过作用于依赖组织因子(tissue factor,TF)的外源性凝血途径对体内凝血系统起调节作用.近年来研究发现,TFPI可对多种疾病的诊断与治疗发挥作用,其中对于冠心病(coro...     

组织因子途径抑制物与冠心病

组织因子途径抑制物主要作用于依赖组织因子的外援性凝血途径,对体内的凝血系统有重要的调节作用.本文就其在冠心病方面的研究情况予以综述.     

组织因子途径抑制物在不稳定斑块发生的作用

动脉粥样硬化最严重的后果之一是血栓形成。已知组织因子是血栓形成的起始因子,组织因子途径抑制物-1是组织因子的生理抑制剂,组织因子途径抑制物-2能抑制细胞外基质降解。通过分子改造,研制出重组人源性长效组织因子途径抑制物(rhlTFPI),其不仅能对抗动脉粥样硬化并发的血栓形成,而且能抑制内膜的增生。探讨rhlTFPI与斑块的稳定性及组织因子途径抑制物-2与细胞外基质降解关系,具有重要的理论意义;有助于今后应用rhlTFPI和重组组织因子途径抑制物-2维持斑块稳定、防治动脉粥样硬化疾病。     

组织因子途径抑制物基因转移对大鼠血管平滑肌细胞凋亡的影响

目的 研究组织因子途径抑制物(TFPI)基因转移对大鼠血管平滑肌细胞(VSMC)凋亡的影响,探讨其抑制血管成形术后再狭窄的机制.方法 在体外培养的大鼠VSMC中分别转染含有人TFPI基因的重组腺病毒(Ad-TFPI)、含β-半乳糖苷酶基因的重组腺病毒(Ad-LacZ)或PBS.通过RT-PCR方法检测外源TFPI基因的表达.细胞计数和MTT法测定细胞生长情况.电镜技术、流式细胞仪和TUNEL法分别检测细胞凋亡情况.结果 基因转移后3 d在VSMC中检测到TFPI mRNA的表达.细胞计数结果显示第1、3、5天各组细胞数无明显差异,而第7天Ad-TFPI组细胞数明显少于Ad-LacZ组和PBS组(P＜0.05).MTT结果显示基因转移后第1、3、5天各组细胞的吸光度值比较差异无统计学意义,而第7天Ad-TFPI组的吸光度值明显低于Ad-LacZ组(P＜0.05)和PBS组(P＜0.01).流式细胞仪检测结果显示基因转移后3、5、7 d Ad-TFPI组细胞早期凋亡结果均高于Ad-LacZ组.基因转移后3 d和7 d,Ad-TFPI组TUNEL阳性率分别为(10.82±1.57)%和(16.95±2.01)%,明显高于Ad-LacZ组(3.46±0.93)%和(5.11±1.29)%(P＜0.05).透射电镜结果显示基因转移后3、5、7 d Ad-TFPI组细胞逐渐出现体积变小、线粒体轻度肿胀、核固缩以及凋亡小体形成,而Ad-LacZ组则无明显改变.结论 TFPI基因转移能够显著诱导体外培养的大鼠VSMC发生凋亡,可能是其抑制血管成形术后再狭窄的机制之一.     

维生素E对组织因子及其抑制物在冠心病中的干预作用

目的 :探讨冠心病 (CHD)的不同类型中血浆组织因子 (TF)及其抑制物 (TFPI)含量的差异变化及维生素E对急性心肌梗死 (AMI)干预作用。方法 :用ELISA方法检测CHD患者 [包括稳定型心绞痛 (SAP)、不稳定型心绞痛 (UAP)、AMI]入院时及治疗 2周时血浆TF、TFPI含量。结果 :①SAP组血浆TF、TFPI水平及TF/TF PI比值均高于正常对照组 ,但差异无统计学意义 (P >0 .0 5 ) ,且治疗前后无明显变化 (P >0 .0 5 )。②UAP组、AMI组血浆TF、TFPI水平及TF/TFPI比值明显高于正常对照组 ,差异有统计学意义 (P <0 .0 1) ,但两组常规治疗前后差异无统计学意义 (P >0 .0 5 )。③AMI加维生素E组干预治疗后TF值显著下降 (P <0 .0 1) ,与正常对照组比较差异无统计学意义 (P >0 .0 5 ) ,TFPI干预治疗后无明显变化 (P >0 .0 5 ) ,而TF/TFPI比值明显降低 (P <0 .0 1)。④各组TF与TFPI呈明显正相关 (P<0 .0 5 ,r=0 .4 32 )。结论 :TF及TFPI在CHD尤其是急性冠状动脉综合征的发生中起重要作用 ,维生素E的干预显著降低患者血浆TF水平 ,降低TF/TFPI比值而对TFPI无影响。     

Clinical significance of expression of tissue factor and tissue factor pathway inhibitor in ulcerative colitis

AIM: To investigate the clinical significance of expression of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in ulcerative colitis (UC).METHODS: Thirty UC specimens taken by colonoscopy from patients with active UC treated at the Department of Pathology, Central Hospital Affiliated to Shenyang Medical College from February 2010 to January 2012 were included in an experimental group, and 30 normal colon tissue samples taken by colonoscopy from non-UC patients were included in a control group. Expression of TF and TFPI in UC and normal colon tissue samples was detected by immunohistochemistry.RESULTS: The positive rate of TF in UC was significantly higher than that in normal colon tissue (63% vs 33%, χ² = 5.41, P < 0.05). The positive rate of TFPI in UC was also significantly higher than that in normal colon tissue (43% vs 17%, χ² = 5.08, P < 0.05).CONCLUSION: Positive rates of TF and TFPI expression in UC are significantly higher than those in normal colon tissue. TF and TFPI may play an important role in the pathogenesis of UC.     

冠心病患者血浆组织因子和组织因子途径抑制物的变化

目的 :通过检测不同类型冠心病 (CHD)患者血浆组织因子 (TF)和组织因子途径抑制物 (TFPI)水平变化 ,探讨其在CHD发病过程中的作用。方法 :以酶联免疫吸附测定法测定CHD患者血浆中TF和TFPI抗原水平。结果 :不稳定型心绞痛 (UAP)和急性心肌梗死 (AMI)患者的血浆TF和TFPI水平与正常对照者和稳定型心绞痛 (SAP)患者相比均有显著性增高 (P <0 .0 5 ) ,以AMI患者尤为明显 (P <0 .0 1) ;UAP和AMI患者的TF PI/TF比值显著降低 (P <0 .0 5 ) ,而SAP患者的上述指标与正常对照者相比 ,其差异均无显著性意义 (P >0 .0 5 )。结论 :UAP和AMI患者TFPI/TF系统失衡 ,标志高凝状态的存在 ;TF和TFPI在这两种类型CHD的发病机制中可能起着重要的作用     

人组织因子途径抑制物腺病毒表达系统的构建及体内表达

目的构建携载人组织因子途径抑制物(tissuefactorpathwayinhibitor,TFPI)基因的重组腺病毒载体,为基因治疗提供实验基础。方法利用基因重组技术,将人TFPI基因连接到穿梭质粒pDC316中,然后将腺病毒骨架质粒pBHGlox△E1,3Cre以及重组穿梭质粒pDC316-TFPI共转染293细胞,并在其中发生Cre重组酶介导的位点,特异性重组及腺病毒包装,扩增后进行滴度测定。将包装成功的携带人TFPI基因的重组腺病毒(Ad-TFPI)转染兔颈动脉,并用携带LacZ报告基因的重组腺病毒(Ad-LacZ)作为对照,3d后RT-PCR、ELISA法检测人TFPImRNA、蛋白的表达。结果得到了携带人TFPI基因的重组腺病毒,包装的病毒蚀斑形成单位(plaqueformationunit,PFU)滴度为7.6×1012/L。在Ad-TFPI转染兔颈动脉后3d,RT-PCR法和ELISA法均检测出TFPI表达,Ad-LacZ转染后未测到人TFPI的表达。结论成功构建了人TFPI腺病毒表达载体,为下一步的基因治疗提供了基础。     

Increased thrombin-activatable fibrinolysis inhibitor and decreased tissue factor pathway inhibitor in patients with hypothyroidism

Various abnormalities of coagulation–fibrinolytic system have been reported in patients with thyroid dysfunction. Several studies indicate that coagulation and fibrinolytic system is disturbed in the patients with hypothyroidism. Also, the influence of hypothyroidism on hemostasis is controversial; both hypocoagulable and hypercoagulable states have been reported. The levels of plasma thrombin-activatable fibrinolysis inhibitor (TAFI) antigen and tissue factor pathway inhibitor (TFPI) have been investigated only once in patients with hypothyroidism. Therefore, the main purpose of this study was to evaluate the profile of coagulation and fibrinolytic parameters including TAFI and TFPI in patients with hypothyroidism. Fifteen patients with untreated hypothyroidism and 15 age-matched healthy controls were included in the study. Factors V(FV), VII (FVII), VIII (FVIII) activities, von Willebrand factor (vWF), protein C, protein S, thrombomodulin (TM), TFPI, and TAFI were measured. The relationships between serum thyroid hormones and these hemostatic parameters were examined. Compared with the control subjects, FVII activity, and TM Ag and TAFI Ag levels were significantly increased in patients with hypothyroidism, whereas FV, FVIII, vWF, protein C and protein S activities, and TFPI Ag levels were significantly decreased. We did not find any significant correlation between serum thyroid hormones and the hemostatic parameters that we measured. In conclusion, we found some important differences in the hemostatic parameters between the patients with hypothyroidism and healthy controls. Increased FVII, TM, and TAFI and decreased FV, FVIII, vWF, protein C, protein S, and TFPI in these patients represent a potential hypercoagulable and hypofibrinolytic state, possible endothelial dysfunction, which might augment the risk for atherosclerotic and atherothrombotic complications. Thus, disturbances of the hemostatic system may contribute to the excess mortality due to cardiovascular disease seen in patients with hypothyroidism.     

Adenovirus-mediated local expression of human tissue factor pathway inhibitor eliminates shear stress-induced recurrent thrombosis in the injured carotid artery of the rabbit.

The main cause of acute coronary syndrome may be recurrent thrombosis, which is initiated by the activation of the extrinsic coagulation pathway. Tissue factor (TF) pathway inhibitor (TFPI) efficiently inhibits an early step in this pathway by the formation of a complex with factor VIIa, TF, and factor Xa. We determined whether local TFPI gene transfer can inhibit thrombosis in an injured artery without inducing systemic side effects. Balloon-injured rabbit carotid arteries were infected with an adenoviral vector that expressed either human TFPI (AdCATFPI) or bacterial beta-galactosidase (AdCALacZ). Two to 6 days after gene transfer, thrombosis was induced by the production of constant stenosis of the artery, and blood flow was measured continuously with an electromagnetic flow probe. A cyclic flow variation, which is thought to reflect the recurrent formation and dislodgment of mural thrombi, was observed in all AdCALacZ-infected arteries as well as in saline-infused arteries. In contrast, no cyclic flow variation was detectable in AdCATFPI-transfected arteries, even in the presence of epinephrine (1 microg. kg-1. min-1 infusion). Prothrombin time, activated partial thromboplastin time, and the ex vivo platelet aggregation induced by either adenosine diphosphate or collagen were unaltered in AdCATFPI-infected rabbits. We found that in vivo TFPI gene transfer into an injured artery completely inhibits the recurrent thrombosis induced by shear stress even in the presence of catecholamine, without affecting systemic coagulation status. Adenovirus-mediated local expression of TFPI may have the potential for the treatment of human thrombosis.