慢性心力衰竭患者血浆组织型纤溶酶原激活物和纤溶酶原激活物抑制物-1含量变化及其临床意义

目的　研究慢性心力衰竭 (CHF)患者血浆组织型纤溶酶原激活物 (t PA)和纤溶酶原激活物抑制物 1(PAI 1)含量的变化及其临床意义。方法　用酶联免疫吸附法 (ELISA)检测 6 0例CHF患者 (CHF组 )和 2 0例健康体检者 (正常对照组 )血浆t PA及PAI 1抗原含量。结果　CHF组血浆t PA和PAI 1平均含量都明显高于对照组 (P<0 .0 1)。CHF患者血浆PAI 1含量增高随心功能恶化而愈加明显。结论　CHF患者纤溶功能明显下降 ,可用血浆t PA、PAI 1含量作为判断病情的参考指标之一。     

缬沙坦对肾小球硬化大鼠纤溶酶原激活物抑制物-1表达影响

目的:探讨缬沙坦对大鼠肾小球硬化细胞外基质降解系统的影响.方法:30只雄性SD大鼠随机分为A,B,C 3组.对B,C 2组大鼠行左肾切除,1周后经尾静脉注射阿霉素,建立肾小球硬化模型.A组行假手术及尾静脉注射等量生理盐水为正常对照.C组给予缬沙坦20 mg/kg,灌胃1次/d;A,B 2组给予2 mL清水灌胃,1次/d.12周后处死大鼠.观察各组大鼠24 h尿蛋白含量及血生化指标和肾脏病理改变,用免疫组织化学方法检测肾脏组织中Ⅳ型胶原、纤溶酶原激活物抑制物-1蛋白表达;用原位杂交方法检测肾脏组织纤溶酶原激活物抑制物-1 mRNA的表达.结果:第6周C组尿蛋白较B组明显降低(P<0.05),第12周血白蛋白较B组明显增高(P<0.05);B组多数肾小球呈节段性硬化,C组肾小球硬化指数明显低于B组(P<0.01);C组Ⅳ型胶原、纤溶酶原激活物抑制物-1在肾脏沉积较B组明显减少(P<0.05);纤溶酶原激活物抑制物-l mRNA表达量明显增加(P<0.05).纤溶酶原激活物抑制物-1及纤溶酶原激活物抑制物-1 mRNA吸光光度值与Ⅳ型胶原均呈正相关.结论:缬沙坦通过降低纤溶酶原激活物抑制物-1在肾脏表达,促进细胞外基质降解而延缓肾小球硬化的进展.     

纤溶酶原激活物抑制物的研究进展

纤溶酶原激活物的快速作用的抑制物(简称PAD 是近几年才证实的一种人类血浆纤溶系统新成分,是调节纤溶活性的重要因子。它目前暂分为三类:PAI-1,PAI-2和蛋白酶结合素(Profease Nexin)。对其生化性质已做了较深入的研究。在临床上,由于PAI 的发现,更新了纤溶活性的概念,实际上血浆纤溶活性的高低是由PAI 与纤溶酶原激活物(t-PA)二者的比值所决定的,许多病人的纤溶活性下降而导致血栓倾向或血栓形成,并非是t-PA 水平的下降,而是由于血浆PAI 的升高这一点对进一步阐明血栓性疾病的发病机制和指导临床纤溶活性测定与血栓疾病的治疗将具有重要意义。     

D-二聚体及与纤溶酶原激活物抑制剂-1比值评价合并糖耐量减低急性冠状动脉综合征患者纤溶受抑的研究

目的 :探讨合并糖耐量减低 (IGT)的急性冠状动脉综合征 (ACS)患者纤溶受抑状态及其对治疗和预后的影响。方法 :非糖尿病 ACS组 39例 ,IGT+ACS组 37例 ,非胰岛素依赖性糖尿病 (NIDDM) +ACS组 36例 ;同时设正常对照组 2 0例 ,检测各组组织型纤溶酶原激活剂 (t PA) ,纤溶酶原激活物抑制剂 1(PAI 1)及D 二聚体的血浆水平 ,并计算 PAI 1/ D 二聚体百分比。观察各组急性心肌梗死 (AMI)患者的临床及预后。结果 :IGT+ACS及 NIDDM+ACS组的 PAI 1显著高于正常对照组和 ACS组 (P均 <0 .0 5 ) ,而 D 二聚体上升幅值则显著低于 ACS组 (P<0 .0 5 ) ,两组 PAI 1/ D 二聚体比值也均显著高于正常对照组及 ACS组(P均 <0 .0 1)。两糖代谢异常组溶栓再通比例显著低于 ACS组 (P<0 .0 1) ,泵衰竭的发生率显著高于 ACS组(P<0 .0 5 )。结论 :IGT的 ACS患者存在纤溶受抑状态。 D二聚体血浆水平结合 PAI 1/ D二聚体比值能反映患者纤溶受抑的状态及预后。     

明胶酶B及其抑制物与大鼠肾脏衰老关系的研究

目的　探讨明胶酶B(MMP 9)及其抑制物 (TIMP 1)在大鼠肾脏衰老过程中的作用。方法　选用 3月、12月、2 4月龄大鼠 ,采用免疫组化技术分别检测MMP 9、TIMP 1、Ⅲ型胶原 (ColⅢ )、纤连蛋白 (FN)、增殖细胞核抗原 (PCNA)及α 平滑肌肌动蛋白 (α SMA)在不同鼠龄大鼠肾组织中的表达。结果　TIMP 1主要表达在肾小球、小管间质及血管 ,并随增龄表达增强 (P <0 0 1) ;MMP 9、主要表达在肾小管上皮细胞 ,随增龄表达无变化 ;ColⅢ主要表达在肾小球及小管间质 ,随增龄表达增强 (P <0 0 1) ;FN主要表达在肾小球及小管间质 ,随增龄表达增强 (P <0 0 1) ;α SMA除正常在血管平滑肌表达以外 ,在 2 4月鼠肾间质纤维化部位、肾小球系膜细胞及增厚的包曼氏囊有阳性表达 ;PCNA在 2 4月鼠肾小球、肾小管的细胞核及肾间质纤维化部位的成纤维细胞核有表达。TIMP 1与肾小球硬化有相关性 (P <0 0 5 )。TIMP 1与小管间质纤维化面积有相关性 (P <0 0 5 )。结论　MMP 9/TIMP 1表达失衡在大鼠肾脏衰老过程中可能起重要作用。     

淋巴瘤患者血浆尿激酶型纤溶酶原激活物及其受体检测的临床意义

尿激酶型纤溶酶原激活物（u—PA）与尿激酶型纤溶酶原激活物受体（u—PAR）是近年特别关注的2个纤溶因子，他们除有纤溶作用外，还参与胞外基质降解、细胞修复和组织重塑等过程。这2个因子与白血病之间的关系有一些报道，但尚未见淋巴瘤方面的临床研究报告。为此，我们检测了淋巴瘤患者血浆u—PA和u—PAR，并对其含量变化的临床意义进行了探讨。     

组织型纤溶酶原激活物、尿激酶型纤溶酶原激活物在单侧输尿管梗阻大鼠不同阶段肾组织中的表达及意义

目的:建立单侧输尿管梗阻大鼠模型,观察组织型纤溶酶原激活物(t-PA)、尿激酶型纤溶酶原激活物(u-PA)在不同时间点肾组织中的表达情况,探讨其不同的表达与肾间质纤维化的关系。方法:实验于2006-05/2007-04在泸州医学院附属医院病理学实验室及传染免疫学实验室完成。72只清洁级雄性SD大鼠,随机分为3组,正常组24只,假手术组24只,模型组24只。各组大鼠于单侧输尿管结扎术后1,4,7,14d取肾组织进行病理分析,观察各组大鼠于单侧输尿管梗阻术后1,4,7,14d的肾小管损伤与肾间质纤维化情况,并采用免疫组织化学技术动态观察t-PA、u-PA在各组的表达情况。实验过程中对动物处置符合动物伦理学标准。结果:参加实验动物72只,中途共死亡5只,均及时补充,最后进入结果分析大鼠为72只。①模型组术后4d,肾脏组织即表现早期纤维化的病理改变,术后第7天,肾小管表现出明显损害,术后14d,肾间质纤维化表现明显。②假手术组较正常组各时间点t-PA、u-PA表达明显减少(P<0.05),模型组t-PA、u-PA在梗阻初期均随梗阻时间延长表达增多,术后第4天表达最多,以后随梗阻时间延长表达逐渐减少。结论:在输尿管梗阻早期,肾组织可能自身代偿分泌合成t-PA、u-PA,在一定程度下有利于延缓肾间质纤维化的进展,随着纤维化的发展,肾小管上皮细胞变性坏死,t-PA、u-PA分泌逐渐减少。     

先天性纤溶酶原激活物抑制剂-1缺乏症的诊断研究

目的　诊断 1例先天性纤溶酶原激活物抑制剂 1(PAI 1)缺乏症患者并研究其基因异常。方法　用发色底物法、酶联免疫吸附实验测定组织型纤溶酶原激活物 (tPA)、α2 纤溶酶抑制剂因子(α2 PI )与PAI 1的活性和 (或 )抗原 ,PCR法与DNA测序分析患者的基因异常 ,用限制性内切酶PshAⅠ对患者和 6 0名正常人PCR产物酶切以排除基因多态性。结果　患者优球蛋白溶解时间 (ELT) 70min ,血浆中加入生理浓度的PAI 1(5 0ng ml)后ELT延长至 12 0min ,PAI 1活性 0 .0 4AU ml,PAI 1抗原 5 .6ng ml,α2 PI活性、F活性及tPA抗原和活性均正常。PCR测序证实患者PAI 1基因外显子 2第 4 3位核苷酸G→A杂合性改变 ,导致信号肽第 15位的丙氨酸突变为苏氨酸。酶切鉴定排除了多态性。结论　报道国内首例PAI 1缺乏症患者 ,其基因改变可能为复合杂合子 ,其中之一为Ala15Thr,该突变为一种国际上尚未报道的新的基因突变。     

去卵巢大白鼠血浆组织型纤溶酶原激活物与其抑制物的变化

测定了去卵巢大白鼠血浆组织型纤溶酶原激活物(t-PA)与纤溶酶原激活物抑制物(PAI)水平。与对照组比较去卵巢组血浆t-PA无显著性差异(P>0.05)。对照组PAI为9.7±1.2AU/ml,去卵巢组PAI为7.8±1.9AU/ml,两组比较有显著差异(P<0.05)。去卵巢组血浆中PAI降低,可能与卵巢激素的减少有关。卵巢激素会促使PAI合成并释放增加,这可能是服用雌激素等避孕药者纤溶活性降低、血液呈高凝状态的重要机制之一。     

肝硬化和肝癌尿激酶型纤溶酶原激活物及受体检测的意义

一些研究表明体液中的尿激酶型纤溶酶原激活物（urokinase type plasminogen activator，u—PA）和尿激酶型纤溶酶原激活物受体（urokinase type plasminogen activator receptor，u—PAR）浓度与癌细胞自身分泌和释放及其浸润转移有关。原发性肝癌是临床上常见的又与乙型肝炎（简称乙肝）后肝硬化有关的癌症。     

消化道肿瘤患者血浆尿激酶型纤溶酶原激活物系统的变化及其意义

目的观察消化道恶性肿瘤患者血浆尿激酶型纤溶酶原激活物(u-PA)及其特异性受体(u-PAR)和纤溶酶原激活物抑制物-1(PAI-1)含量的变化及其与肿瘤转移和预后的关系.方法用酶联免疫吸附测定(ELISA)法测定43例消化道恶性肿瘤患者和21例正常人血浆中u-PA、u-PAR和PAI-1含量.结果食管癌、胃癌和结肠癌患者的血浆u-PA、u-PAR和PAI-1含量均显著升高(P<0.05～0.01);肿瘤组中,中、晚期组u-PA显著高于早期组(P＜0.05),已转移组u-PA、u-PAR和PAI-1较未转移组显著升高(P<0.05～0.01).结论消化道恶性肿瘤患者血浆中u-PA、u-PAR和PAI-1含量不同程度升高,并与肿瘤转移和预后相关.     

The effect of levonorgestrel-releasing intrauterine system use on menstrual blood loss and the hemostatic, fibrinolytic/inhibitor systems in women with menorrhagia

BACKGROUND: Menorrhagia is known to be associated with uterine fibroids, adenomyosis, pelvic infections, endometrial polyps and clotting defects. A viable alternative therapy to hysterectomy should alleviate heavy menstrual blood flow and consequently improve the quality-of-life measures in women presenting with menorrhagia. The levonorgestrel-releasing intrauterine system (LNG-IUS) ranks higher than medical treatments in terms of efficacy, comparable improvements in quality of life and psychological well-being. OBJECTIVE: The purpose of the study was to determine the effects of 6 months of LNG-IUS use on menstrual blood loss and the hemostatic, fibrinolytic/inhibitor systems in blood and the endometrium in women with menorrhagia with known pathologic causes. PATIENTS AND METHODS: Samples from 41 women were analyzed. Hemoglobin, hematocrit, thrombelastography, tissue-type plasminogen activator (t-PA), urokinase plasminogen activator (u-PA), u-PA receptor (u-PAR), plasminogen activator inhibitor-1/2 (PAI-1/2), D-dimer and von Willebrand factor (VWF) were determined, and t-PA, u-PA and PAI-1/2 were also determined in endometrial tissue extracts. Results: Menorrhagia was reduced in 89% of women by 3 months; by 6 months all women had no menorrhagia, and 39% of women had become amenorrhoeic. Hemoglobin and hematocrit levels showed improvement, and reached normal reference levels by 6 months. There were no systemic changes in the fibrinolytic/inhibitor systems and VWF, except for a decreased u-PAR level. However, in the endometrium, significant elevations in PAI-1/2 together with u-PAR levels were seen at 6 months. CONCLUSIONS: The slow levonorgestrel-release intrauterine device use results in high expression of fibrinolytic inhibitors (PAI-1/2) and upregulated u-PAR expression in the endometrium. Systemic hemostasis was not significantly altered. The study demonstrated that LNG-IUS is highly effective in the treatment of menorrhagia with known pathologic causes.     

老龄大鼠脑缺血-再灌注微血管基底膜损伤变化及与纤溶酶原激活系的关系

目的研究老龄大鼠脑缺血一再灌注（I／R）不同时期微血管基底膜损伤与纤溶酶原激活系的关系。方法采用大脑中动脉阻塞（MCAO）线栓法制备大鼠局灶性脑I／R模型，将大鼠分为青年假手术组、老龄假手术组、青年模型组和老龄模型组，其中模型组又分为I3h，I／R6、12、24、72和144h各时间点组。采用免疫组化、酶谱与反向酶谱分析等方法测定各脑微血管结构、基底膜Ⅳ型胶原、层连蛋白（LN）和纤溶酶原激活系的变化。结果与青年假手术组比较，老龄假手术组Ⅳ型胶原、LN表达增强。随着I／R时间的延长，老龄与青年大鼠基底膜成分Ⅳ型胶原和LN表达递减；组织型纤溶酶原激活剂（t-PA）、尿激酶型纤溶酶原激活剂（u-PA）、纤溶酶原激活物抑制剂（PAI-1）表达水平呈先增强后降低的趋势。与青年模型组相同时间点比较，老龄模型组Ⅳ型胶原（I3h、I／R6h、I／R12h）、LN（I3h、I／R6h～24h）、t-PA（I／R6h～24h）、u-PA（I／R12h～144h）表达增强，PAI-1（I／R12h、I／R24h）表达降低，差异均有显著性。另外，PAI-1的反向酶谱分析比较，量的变化与免疫表达规律基本一致。结论随着增龄，大鼠脑微血管基底膜成分Ⅳ型胶原、LN增加。在脑I／R微血管基底膜损伤方面，老龄大鼠较青年严重，其损伤的特点与纤溶酶原激活系变化有关。     

Disruption of plasminogen activator inhibitor-1 gene enhances spontaneous enlargement of mouse airspace with increasing age

Plasminogen activator inhibitor-1 (PAI-1) is the most effective protease inhibitor in the fibrinolysis system, and plays an important role in the remodeling of the extracellular matrix. We therefore explored whether PAI-1 is involved in the change of lung structure with increasing age. PAI-1 gene knockout mice and wild-type mice were sacrificed at age 3 weeks, 3 months, 6 months and 15 months for histopathology analysis, and assessed the relationship between PAI-1 and the change in lung structure with age. Six-month-old mice were chosen for further studies. Elastin in the lung was detected using Weigert staining. We measured the expression of matrix metalloproteinase-12 (MMP-12) that is a major protease in elastin degradation by real time PCR and immunostaining. Transforming growth factor-β1 (TGF-β1) expression was measured by western blot analysis. PAI-1 gene knockout mice showed significant increases in alveolar size with increasing age and damaged alveolar structure at the age of 15 months, compared with wild-type mice. At the age of 6 months, elastin protein was decreased in the lungs of PAI-1 gene knockout mice. PAI-1 null mice had higher MMP-12 mRNA expression, and lower expression level of active TGF-β1 in the lung. Taken together, these results indicate that the emphysema-like change attributed to PAI-1 deficiency might be facilitated with increased MMP-12 expression that accelerates elastin degradation in mice lungs, and TGF-β1 might be involved in the modulation of this process.     

脑血管病患者血小板、纤溶活性、内皮细胞合成物测定的意义

目的：探讨血浆中血栓素 Ｂ_２（ＴＸＢ_２）,血小板α颗粒胰蛋白－１４０（ＧＭＰ－１４０）, ６－酮前列腺素１α（６－Ｋ－ＰＧＦ１α）,内皮素（ＥＴ）、组织型纤溶酶原激活剂（ｔＰＡ）、纤溶酶原激活剂抑制物（ＰＡＩ）活性,在脑动脉硬化、脑血栓发病过程的作用及临床意义。方法：ＴＸＢ_２、Ｇｍｐ－１４０、６－Ｋ－ＰＧＦ１α、ＥＴ采用放射免疫法（ＲＩＡ）、ｔＰＡ、ＰＡＩ采用发色底物法分别测定脑动脉硬化,脑血栓和正常对照组的含量,并进行分析。结果：①脑动脉硬化组,脑血栓组血小板指标ＴＸＢ_２,ＧＭＰ－１４０水平明显高于正常组（Ｐ均＜０．０１）;②内皮细胞合成物６－Ｋ－ＰＧＦ１α、ＥＴ含量脑血栓组明显低于和高于正常组（Ｐ均＜０．０１）,脑动脉硬化组无明显差异（Ｐ均＞０．０５）。 ③纤溶指标 ｔＰＡ, ｔＰＡ／ＰＡＩ两组脑血管病血浆水平明显低于正常组。 ＰＡＩ水平虽有增高,但无明显差异（Ｐ＞０．０５）。结论：脑血管疾病存在血小板活化,内皮细胞受损、纤溶活力低下,这些改变可能参与此二类疾病发病过程。故须针对以上改变对患者采用相应治疗措施。     

Renal synthesis of urokinase type-plasminogen activator, its receptor, and plasminogen activator inhibitor-1 in diabetic nephropathy in rats: modulation by angiotensin-converting-enzyme inhibitor

Plasmin is an important factor in the degradation of extracellular matrix. In the study reported here we examined the expression of plasminogen-activator inhibitor-1 (PAI-1), urokinase-type plasminogen activator (uPA), and uPA receptor (uPAR), as well as the relevance of such expression to the production of type IV collagen, a major component of extracellular matrix, in the renal tissue of rats with streptozotocin-induced diabetes. Because angiotensin II is involved in the synthesis of PAI-1 and uPA, we also examined the effect of benazepril, an angiotensin-converting-enzyme inhibitor, on the expression of PAI-1, uPA, and uPAR messenger RNAs (mRNAs) and type IV collagen protein. Rats with streptozocin-induced diabetes-some untreated and some treated with 30 mg/L benazepril-and nondiabetic control rats were sacrificed at 4, 12, or 24 weeks after induction of diabetes. We examined the expression of PAI-1, uPA, and uPAR mRNAs through the use of in situ hybridization and that of type IV collagen by means of immunohistochemical methods. In control rats, we detected weak signals for PAI-1, uPA, and uPAR mRNAs in glomeruli. Diabetic rats exhibited high levels of expression of PAI-1, uPA, and uPAR mRNAs and type IV collagen protein, mainly in mesangial cells. These mRNAs were synthesized in various renal cells (epithelial, mesangial, and endothelial cells and Bowman's capsule). Benazepril inhibited increases in all 3 mRNAs, especially in the mesangium; reduced type IV collagen expression; and attenuated mesangial expansion. Our results indicated that altered expression of PAI-1, uPA, and uPAR in diabetic nephropathy was associated with mesangial expansion and that the beneficial effects of ACE-I may be at least associated with such expression.     

Glucosamine sulfate suppresses the expressions of urokinase plasminogen activator and inhibitor and gelatinases during the early stage of osteoarthritis

BACKGROUND: Glucosamine sulfate may have an ex vivo inhibitory effect on the plasminogen activator (PA)/plasmin system and gelatinases expression during the early development of osteoarthritis (OA). METHODS: We compared the levels of urokinase-type PA (u-PA), PA inhibitor-1 (PAI-1) and gelatinases (matrix metalloproteinase-2 and -9 [MMP-2 and -9]) in a series of chondral, meniscal, and synovial cultures of early OA after treatment with or without glucosamine sulfate. RESULTS: Gelatin zymography revealed that glucosamine sulfate could suppress MMP-2 secretion in chondral, meniscal and synovial cultures and also decrease MMP-9 production in synovial and meniscal cultures. ELISA data also showed the suppressive effects of glucosamine sulfate on u-PA and PAI-1 production in synovial cultures at 48 h. CONCLUSIONS: Our data suggest that one of the therapeutic effects of glucosamine sulfate is to down-regulate the expressions of u-PA, PAI-1, MMP-2 and MMP-9 that underlie the destruction of articular cartilage in the early stage of OA, and therefore to delay the joint failure.     

游离脂肪酸对肾小管上皮细胞TIMP-1、PAI-1表达及细胞外基质分泌的影响

目的探讨游离脂肪酸（Free fatty acids，FFAs）对人肾小管上皮细胞（renal tubular epithelial cells，RTECs）细胞外基质降解酶抑制剂[tissue inhibitor of metalloproteinase 1（TIMP-1）、plasminogen activator inhibitor 1（PAI-1）]表达及细胞外基质[纤维连接蛋（fibronectin，FN）、胶原]分泌的影响。方法用去脂牛血清白蛋白（defatty bovine serum albumin，d-BsA）为油酸（Oil acids，OA）载体，用不同浓度OA刺激培养RTECs，以不加d-BSA及只加普通培养液的细胞作对照组，应用反转录多聚酶链反应（RT-PCR）检测RTECs TIMP-1mRNA、PAI-1mRNA的表达水平，酶联免疫吸附法（FLISA法）、羟脯氨酸消化法分别检测培养上清FN、胶原的蛋白水平。结果d-BSA组与空白对照组RTECs的细胞外基质降解酶抑制剂（TIMP-1mRNA、PAI-1mRNA）表达及细胞外基质蛋白（FN、胶原）分泌比较差异无显著性（P＞0．05），而不同浓度OA组与空白对照组比较差异有显著性（P＜0．05）；且随着OA浓度增加，对RTECs TIMP-1mRNA、PAI-1mRNA表达以及对FN、胶原蛋白分泌的促进作用有增强趋势。结论FFAs通过促进细胞外基质（FN、胶原）的生成及抑制其降解，使其沉积在肾间质，进而在肾小管间质纤维化发生发展中起着重要的作用。     

PAI-1蛋白在子宫内膜异位症组织的表达及意义

目的：探讨PAI-1蛋白在异位内膜、在位内膜厦对照组内膜的表达及意义。方法：应用免疫组化方法检测PAI-1蛋白在40例内异症组异位内膜及在位内膜，30例对照组子宫内膜的表达。结果：①PAI-1蛋白表达于异位内膜、在位内膜、对照组内膜的腺上皮细胞及间质细胞胞浆；②PAI-1蛋白在异位内膜厦对照组内膜间质细胞表达均高于腺上皮细胞（P〈0．05），而于在位内膜腺上皮细胞与间质细胞表达比较无统计学意义（P〉0．05）；③PAI-1蛋白在间质细胞表达异位内膜高于在位内膜及对照组内膜（P〈0．05），而在腺上皮细胞表达异住内膜、在位内膜、对照组内膜三者之间比较无统计学意义（P〉0．05）；④PAI-1蛋白在血管内皮细胞呈阳性表达。结论：PAI-1蛋白在异位内膜间质细胞高表达可能促使其转移、黏附、侵袭、生长，导致子宫内膜异住症的发生发展。