人正常胚胎和葡萄胎绒毛中纤蛋白溶酶原激活与抑制因子及其尿激酶？…

本研究应用原位杂交技术检测人正常胚胎及葡萄胎绒毛中纤蛋白溶酶原激活与抑制因子（ｔＰＡ，ｕＰＡ，ＰＡＩ－１）及其尿激酶受体（ｕＰＡ－Ｒ）的表达。结果表明，人正常胚胎绒毛的合体滋养层和细胞滋养层细胞中均含有ｔＰＡ、ｕＰＡ、ＰＡＩ－１及ｕＰＡ－Ｒ ｍＲＮＡ，葡萄胎绒毛中ｔＰＡ、ＰＡＩ－１及ｕＰＡ－Ｒ ｍＲＮＡ表达明显增强。提示：（１）ＰＡ－ＰＡＩ的协同作用在维持妊娠期间正常的纤蛋白溶解可能是重要的；（２     

纤溶酶原激活及抑制因子在人蜕膜腺体与间质细胞中的生成及调节

为了探讨人早孕蜕膜腺体与间质细胞离体培养下释放活性纤蛋白溶酶原激活及抑制因子（ＰＡ，ＰＡＩ）的平衡关系与激素调节。应用琼脂糖纤蛋白铺盖及反向铺盖技术，测得腺体及间质细胞均仅释放尿激酶型ＰＡ（ＵＰＡ），而不释放组织型ＰＡ（ｔＰＡ），并同时生成Ⅰ型ＰＡ抑制因子（ＰＡＩ－１）。雌二醇（Ｅ２）及ＲＵ４８６刺激ｕＰＡ、抑制ＰＡＩ－１活性，孕酮（Ｐ）的作用则相反，提示Ｐ在人早孕蜕膜中的作用之一是使ＰＡ－ＰＡ１的动态平衡趋于纤溶活性受抑的安静状态，而ＲＵ４８６激活蜕膜纤溶过程，从而增强细胞外基质蛋白水解，可能是其抗早孕作用机理之一。     

米非司酮对早孕蜕膜中纤蛋白溶酶原激活及抑制因子、及其尿激酶受体的分布与表达的影响

为了探讨米非司酮抗早孕作用是否影响了细胞外基质蛋白水解，作者应用免疫组织化学及原位杂交技术检测正常早孕及米非司酮流产蜕膜中ｔＰＡ、ｕＰＡ、ＰＡＩ－１及ｕＰＡ－Ｒ抗原及相应ｍＲＮＡ。结果表明ｕＰＡ－Ｒ及其ｍＲＮＡ在米非司酮流产蜕膜组织的两种主要组分，尤其是腺体细胞中明显增加。提示米非司酮刺激ｕＰＡ－Ｒ生成增加可激活细胞外基质蛋白水解，因而与抗早孕作用可能有关。     

纤溶酶原激活物抑制物1与肝细胞癌

目的研究纤溶酶原激活物抑制物１（ＰＡＩ１）在肝细胞癌（ＨＣＣ）蛋白和ｍＲＮＡ水平的表达及其与ＨＣＣ生物学特性的关系。方法取ＨＣＣ石蜡标本４８例,肝良性肿瘤石蜡标本１２例（对照组）做免疫组化染色;液氮冻存ＨＣＣ标本２０例,肝血管瘤５例（对照组）做免疫印迹杂交。结果肝癌细胞与癌周细胞及对照组肝细胞相比,ＰＡＩ１抗原蛋白和ｍＲＮＡ表达显著升高,差异有显著意义,Ｐ值分别＜００１和＜０．０５。术后２年内死亡病例与生存病例相比,ＰＡＩ１阳性率有显著意义的升高,Ｐ＜００５。ＰＡＩ１和纤溶酶原激活物（ｕＰＡ）及其受体（ｕＰＡＲ）同时阳性患者与同时阴性患者相比,前者侵袭性病例较后者升高有显著性意义（Ｐ＜００５）。结论ＨＣＣ中ＰＡＩ１蛋白和ｍＲＮＡ表达明显增高。ＰＡＩ１与ＨＣＣ浸润转移和预后密切相关。     

周围神经挤压伤后纤溶成分表达的变化

目的 观察大鼠坐骨神经挤压伤后纤溶成分中组织型纤溶酶原激活剂（ｔ－ＰＡ）和纤溶酶原激活剂抑制因子１（ＰＡＩ－１）蛋白表达的变化,了解ｔ－ＰＡ、ＰＡＩ－１在周围神地操作后修复过程中的作用。方法 取２１只Ｗｉｓｔａｒ大鼠,分别造成坐骨神经挤压伤模型。在术后６ｈ、１ｄ、３ｄ、７ｄ、１４和２１ｄ分别取材进行组织学、免疫组化观察。结果 术后３ｄ时神经断端回缩球处的ｔ－ＰＡ抗原呈阳性反应。７ｄ时损伤进行组织学     

米非司酮对早孕蜕膜中纤蛋白溶酶原激活及抑制因子,及其尿激酶受 …

为了探讨米非司酮抗早孕作用是否影响了细胞外基质蛋白水解。作者应用免疫组织化学及原位杂交技术检测正常早孕及米非司酮流产蜕膜中ｔＰａ、ｕＰＡ、ＰＡＩ及ｕＰＡ－Ｒ抗原及相应ｍＲＮＡ，结果表明ｕＰＡ－Ｒ及其ｍＲＮＡ在米非司酮流产蜕膜组织的两种主要组分，尤其是腺体细胞中明显增加。提示米非司酮刺激ｕＰＡ－Ｒ生成增加可激活细胞外基质蛋白水解，因而与抗早孕作用可能有关     

黄腐酸钠对实验性糖尿病大鼠肾脏的保护作用

目的研究黄腐酸钠对糖尿病大鼠肾脏病变的保护作用及其机制。方法观察黄腐酸钠对肾小球形态，肾小球基底膜超微结构，尿白蛋白排出率的影响及血浆组织型纤溶酶原激活剂（ｔＰＡ）、纤溶酶原激活剂抑制物（ＰＡＩ１）活性改变。结果糖尿病大鼠血浆ｔＰＡ活性明显降低，而ＰＡＩ１活性升高。黄腐酸钠明显降低糖尿病大鼠尿白蛋白，抑制肾小球肥大，延缓肾小球基底膜增厚及足突融合，显著提高血浆ｔＰＡ活性而降低ＰＡＩ１活性。结论黄腐酸钠对糖尿病肾病具有保护作用。此作用可能与提高血浆纤溶活性有关。     

倍美力对绝经后妇女纤溶活性的影响

为了探讨激素替代治疗（ＨＲＴ）的心血管保护机制，观察了倍美力对绝经后妇女纤溶活性的影响。４８例绝经后妇女分为３组：安慰剂组１０例；单用倍美力组（Ｅ组）１７例；倍美力、孕激素合用组（Ｅ＋Ｐ组）２１例，并以２０例绝经前妇女作为对照。采用发色底物法测定了ＨＲＴ治疗前及治疗３个月后血浆组织纤溶酶原激活剂（ｔＰＡ）及其抑制因子（ＰＡＩ）活性。结果：绝经后妇女治疗前ＰＡＩ活性明显高于对照组（Ｐ＜０．０１）。安慰剂组治疗前后ｔＰＡ及ＰＡＩ均无显著性变化（Ｐ＞０．０５）。Ｅ及Ｅ＋Ｐ组治疗３个月后ＰＡＩ活性明显减低，ｔＰＡ活性明显升高（Ｐ＜０．０１），两组比较，治疗前后ｔＰＡ及ＰＡＩ活性均无显著性差异。认为ＨＲＴ可通过改善绝经后妇女纤溶活性保护心血管。     

肢体负压对后肢动脉闭塞犬纤溶酶原激活物及其抑制因子1？…

目的 探讨肢体负压对犬纤溶系统的影响。方法 通过建立后肢动脉闭塞犬模型，检测肢体负压（ＬＮＰ）前后外周血纤溶酶原激活物（ｔ－ＰＡ）及其抑制物（ＰＡＩ－１）活性、静脉血氧分压（ＰＯ２）和ＰＨ值的变化。结果 ＬＮＰ治疗后实验组ｔ－ＰＡ活性、ＰＯ２、ＰＨ值较治疗前明显升高，ＰＡＩ－１活性显著下降（Ｐ〈０．０１），与对照组比较也有显著性差异（Ｐ〈０．０１）。结论 ＬＮＰ可提高局部纤溶活性，促进血栓溶解和再     

纤溶酶原激活、抑制系统及其在女性生殖中的作用

纤溶酶原激活、抑制系统及其在女性生殖中的作用朱艳陈贵安刘以训纤维蛋白溶解与抑制系统主要包括无活性的纤溶酶原（Ｐｌａｓｍｉｎｏｇｅｎ）、纤溶酶（ｐｌａｓｍｉｎ）、纤溶酶原激活因子（ｐｌａｓｍｉｎｏｇｅｎａｃｔｉｖａｔｏｒ，ＰＡ）及纤溶酶原激活因子抑制因...     

纤维蛋白对肾小球内皮细胞表达纤溶酶原激活物及纤溶酶原激活物抑制物的作用

目的：观察体外培养人肾小球内皮细胞（GEC）表面原位形成的纤维蛋白对GEC表达纤溶酶原激活物及纤溶酶原激活物抑制物（PA／PAI）的影响。方法：应用逆转录聚合酶链反应（RT－PCR）,酶谱分析法与反向酶谱法分别在基因转录水平与蛋白质活性水平上检测纤维蛋白对GEC表达tPA,uPA gn PAI-1r 作用,纤维蛋白平板法检测纤维蛋白对GEC PA／PAI系统的综合效应,结果：纤维蛋白能够明显促进tPA,uPA与PAI－1的mRNA表达上调,无血清RPMI 1640培养下的GEC几乎检测不到PAI知性,但可检测到PAI－1的活性。纤维蛋白能够浓度依赖性刺激GEC tPA与uPA活性增加以及PAI01的活性增加,呈浓度依赖性与时间依赖性,相同剂量的纤维蛋白原与纤维蛋白的作用相似,放线菌酮与放线菌素D均可抑制纤维蛋白上调GEC表达tPA,uPA与PAI的作用,纤维蛋白平板法显示,纤维蛋白对GEC PA／PAI系统的综合效应是以升高PA活性为主,其活性能够被抑肽酶完全阻断。结论：肾脏局部毛细血[管内沉积的纤维蛋白可能通过对GEC PA／PAI系统的调节发挥其病理作用。     

完全性葡萄胎绒毛凋亡与基因表达的相关研究

目的 :探讨完全性葡萄胎绒毛凋亡、增殖活性与 Bcl-2、Bax、Fas、Fas-L及增殖细胞核抗原 ( PCNA)基因表达的相关性。方法 :收集正常早孕绒毛标本及近一年归档的完全性葡萄胎绒毛蜡块 ,原位末端标记法 ( TUNEL)进行细胞凋亡的组织学检测 ,免疫组织化学法测定 Bcl-2、Bax、Fas、Fas-L、PCNA的分布与含量 ,原位杂交法测定 Fas、Fas-Lm RNA的分布与含量。结果 :正常早孕绒毛有少量凋亡细胞 ,主要分布于合体滋养细胞 ,Bcl-2、PCNA蛋白仅在细胞滋养细胞表达且含量丰富 ,Bax、Fas、Fas-L蛋白在合体滋养细胞表达且含量低下。完全性葡萄胎绒毛合体滋养细胞凋亡较正常早孕明显增多 ,Fas、Fas-L 蛋白及其 m RNA不仅在合体滋养细胞而且在细胞滋养细胞表达均明显增强 ,Bcl-2、Bax蛋白在两种细胞中表达也增强 ;PCNA蛋白仅见于细胞滋养细胞 ,含量丰富。结论 :完全性葡萄胎绒毛凋亡细胞增多 ,主要经 Fas/ Fas-L 转录与翻译水平介导 ,表现为高凋亡与高增殖共存现象 ,可能与其无控生长的特性相关。     

Components of the plasminogen activator system and their complexes in renal cell and bladder cancer: comparison between normal and matched cancerous tissues

OBJECTIVE

To analyse and compare the concentration of plasminogen activator (PA), urokinase‐type PA (uPA), tissue‐type PA (tPA), PA inhibitor (PAI)‐1 and PAI‐2, and the complexes uPA‐PAI‐1 and tPA‐PAI‐1 and calculated uPA and tPA uncomplexed with PAI‐1 (‘free’) in urothelial cell carcinoma and matched benign urothelium, and in renal cell carcinoma (RCC) and matched benign renal tissue.

PATIENTS AND METHODS

Tissue samples were obtained during cystectomy (33 patients) and nephrectomy (55), and specific enzyme‐linked immunosorbent assays were used to assess the PA components in extracts of these tissues.

RESULTS

Tissue levels of uPA‐PAI‐1 and tPA‐PAI‐1, but also PAI‐1 itself, were greater in tumorous bladder and kidney tissue than in matched normal tissue (by 1.5–7.8 times). Free tPA was clearly lower in tumour tissue (by 0–0.12‐fold). In bladder cancer, but not in RCC, levels of uPA (15.8‐fold) and free uPA (16.4‐fold) were greater in tumour tissue. Free uPA levels were less in RCC (0.41‐fold). For both normal bladder and kidney tissue, there was no clear correlation between uPA‐PAI‐1 complex and either component. However, the formation of tPA‐PAI‐1 complexes in normal bladder and kidney tissue was primarily determined by PAI‐1. Interestingly, in tumour tissues there was a strong, significant correlation between complex levels and both components.

CONCLUSION

RCC and bladder cancer show distinct profiles of components of the PA system. This study provides a basis for further studies into both the (patho)physiological role of the PA system in these tumours, and into a possible relation with tumour progression and prognosis, and as target for therapy.     

尿激酶型纤溶酶原激活物及其受体高表达增强血管收缩性重塑和内膜增生的研究

目的　探讨血管壁中尿激酶型纤溶酶原激活物 (uPA)和其受体 (uPAR)表达与血管收缩性重塑和内膜增生的相关性。方法　建立兔髂动脉粥样硬化再狭窄模型 ,比较内膜和中膜层的uPA、uPAR表达及其与内膜面积和血管重塑指数的相关性。结果　内膜层uPA、uPAR的抗原和mRNA表达明显强于中膜层 (P <0 .0 1) ,血管壁内膜uPA、uPAR的抗原和mRNA表达水平与血管重塑指数呈负相关 (r =-0 .870 0 7,P =0 .0 10 9;r =-0 .860 3 8,P =0 .0 13 0和r =-0 .845 5 5 ,P =0 .0 165 ,r =-0 .862 40 ,P =0 .0 12 5 ) ,与内膜面积呈正相关 (r=0 .92 0 0 0 ,P =0 .0 3 3 0 ;r=0 .90 772 ,P =0 .0 0 47和r=0 .92 14 9,P =0 .0 0 3 2 ;r =0 .90 614 ,P =0 .0 0 49) ,血管重塑指数和血管内膜面积与中膜uPA、uPAR表达无明显相关性。结论　内膜层uPA、uPAR高表达增强了血管重建后血管壁收缩性重塑和内膜增生。     

Expression of urokinase plasminogen activator and its receptor during acute renal allograft rejection

BACKGROUND: In inflammation, urokinase plasminogen activator (uPA) and its receptor (uPAR) play an important role in fibrinolysis and in activation and chemotaxis of neutrophils and lymphocytes. Moreover, the uPA/uPAR system is involved in processes that affect turnover of the extracellular matrix (ECM). The aim of this study was to determine the local and systemic release of uPAR, and the expression of uPA and uPAR in renal tissues during acute renal allograft rejection. METHODS: Blood, urine, and tissue samples were collected from 33 patients diagnosed with acute allograft rejection and from 14 transplant patients without rejection. From 10 healthy volunteers, blood and urine were collected as a control. In urine and blood samples, the levels of uPAR were determined by enzyme-linked immunosorbent assay (ELISA). Immunostaining and in situ hybridization for uPA and uPAR were performed on renal biopsies. RESULTS: uPAR was detectable at low levels in serum and urine of healthy volunteers and was increased in nonrejecting allograft recipients. Serum and urine levels of uPAR were higher in transplant recipients with rejection compared to nonrejectors. The urine and serum levels of uPAR correlated with the renal function. Immunostaining and in situ hybridization showed an up-regulation of both uPA and uPAR in rejection biopsies. Nonrejected grafts displayed no expression of uPA and uPAR by immunostaining, or of uPAR by in situ hybridization. uPA was detected in a limited number of tubular epithelial cells by in situ hybridization. During rejection, lymphocytes as well as tubular epithelial cells showed uPA and uPAR expression. In the vascular types of rejection, strong expression of uPA was also seen in the entire vessel wall, while uPAR was expressed by the endothelium. CONCLUSION: This study shows that (1) uPA and uPAR are up-regulated during acute renal allograft rejection; (2) uPAR levels in urine and serum correlate with serum creatinine levels, and (3) uPA and uPAR are produced by inflammatory cells, tubular epithelium, and vascular endothelium during acute renal allograft rejection.     

IL-8-stimulated expression of urokinase-type plasminogen activator in human skin and human epidermal cells

Extracellular matrix (ECM) remodeling is essential for normal development and tissue repair. Although many roles for extracellular proteinases in the breakdown of ECM have been established, the regulations of these proteinases in human tissue are not fully understood. Inflammatory cytokines have been implicated in the regulation of several matrix metalloproteinases. To determine whether these mediators have a similar effect on fibrinolysis and the remodeling of the fibrin provisional matrix, we examined the role of cytokines on the regulation of urokinase-type plasminogen activator (uPA) and tissue-type plasminogen activator (tPA) in human skin. In this report, we show that interleukin-8 (IL-8), but not other cytokines tested, is a potent inducer of the 38-kDa uPA in organ-cultured human skin. In addition, the uPA inhibitor, PAI-1, was not affected by IL-8. When primary epidermal human keratinocytes were treated with IL-8, 55-kDa pro-uPA was significantly induced in the conditioned medium. The mRNA expression of uPA in the keratinocytes was found to be constitutively elevated and was not affected by IL-8. To support such a notion, activation of the 5'-flanking promoter of the human uPA gene was measured using the CAT reporter assay. Consistent with the results of mRNA measurement, the promoter is constitutively active in keratinocytes and is not affected by IL-8. In contrast, the promoter construct is neither active in the dermal fibroblasts nor stimulated by the cytokine. This differential transactivation of uPA gene in these cells indicates that keratinocyte-specific factors may govern the basal expression of the gene. These results indicate a complex regulation of uPA expression in epidermal cells.     

人精子tPA和PAI-1的测定及精子上tPA的定位研究

目的 :人精子上组织型纤溶酶原原激活因子 (tPA)的分布及精子中tPA、纤溶酶激活物抑制因子 (PAI 1)活性研究。　方法 :应用底物显色法测定 6 1例人精子中tPA及其PAI 1的活性 ,其中正常对照组 2 0例 (已婚生育男子 ,经 2次精液分析正常者 ) ,少弱畸精症组 41例 ;用免疫组化法进行精子表面膜上tPA定位分析。　结果 :正常对照组精子中tPA和PAI 1的活性分别为 (0 2 4± 0 0 4)IU/ 10 6精子和 (0 2 3± 0 0 3)AU/ 10 6精子 ,少弱畸精症组的tPA和PAI 1活性分别为 (0 12± 0 0 3)IU/ 10 6精子和 (0 35± 0 0 9)AU/ 10 6精子 ,经分析二者之间有极显著差异 (P <0 0 1) ;tPA主要分布在精子的顶体膜、赤道板、颈部和尾部。　结论 :正常健康人精子中的tPA和PAI 1活性有别于弱精症者 ,提示精子上的tPA和PAI 1活性可能与精子活力、精子形态以及精子数有关 ;精子的顶体膜、赤道板、颈部及尾部存在着tPA ,推测其与精子运行、获能等有关。