环孢素对足细胞尿激酶型纤溶酶原激活剂受体表达的影响

目的观察环孢素对脂多糖诱导的足细胞尿激酶型纤溶酶原激活剂受体(uPAR)表达的影响。方法建立体内脂多糖诱导小鼠蛋白尿模型和体外脂多糖足细胞损伤模型,2种模型均分为正常对照组、脂多糖组和脂多糖+环孢素组。通过测量尿蛋白-尿肌酐比值观察各组小鼠尿蛋白的变化情况,通过免疫荧光激光共聚焦、RT-PCR及流式细胞术观察各组小鼠肾组织及足细胞uPAR基因和蛋白水平。结果脂多糖组小鼠尿蛋白-尿肌酐比值高于正常对照组(P<0.01),脂多糖+环孢素组低于脂多糖组(P<0.05)。脂多糖+环孢素组小鼠肾组织中uPAR的表达弱于脂多糖组。足细胞脂多糖组uPAR蛋白荧光信号明显强于正常对照组,给予环孢素后荧光信号减弱。足细胞脂多糖+环孢素组uPAR蛋白表达率和Plaur mRNA为(20.10±10.70)%和0.872±0.167,均低于脂多糖组[(65.17±13.04)%和1.341±0.287,P<0.05],与正常对照组无显著差异[(18.06±7.78)%和1.025±0.167,P>0.05]。结论环孢素可能通过抑制足细胞uPAR的表达,从而稳定和保护足细胞起到降蛋白尿作用。     

尿激酶型纤溶酶原激活剂及其受体在胰腺癌中表达的意义

目的探讨尿激酶型纤溶酶原激活剂(urokinaseplasminogenactivator,uPA)、尿激酶型纤溶酶原激活剂受体(urokinaseplasminogenactivatorreceptor,uPAR)在胰腺癌中表达的意义。方法用反转录聚合酶链反应(RT鄄PCR)和免疫组织化学方法分别检测uPA、uPAR在85例胰腺癌和10例胰腺炎组织中mRNA和蛋白抗原的表达。HPIAS图像分析各组标本相对密度,t检验各组相对密度差异,字2检验uPA、uPAR与胰腺癌肿瘤临床病理之间的相关性。结果uPA、uPARmRNA和蛋白抗原在胰腺炎组织中低表达,胰腺癌组织中uPA、uPARmRNA和蛋白抗原的高表达,胰腺癌中伴有转移者uPA、uPAR的mRNA和蛋白抗原的表达明显高于无转移者。uPA、uPAR的表达与胰腺癌的转移程度高度相关。结论uPA、uPAR是反映胰腺肿瘤进展和生物学特性的指标,是临床上判断预后的良好指标之一     

肾炎患者外周血单个核细胞尿激酶型纤溶酶原激活剂受体及相关细胞因子的表达

目的　探讨肾炎患者外周血单个核细胞尿激酶受体及相关细胞因子的表达。方法　采用流式细胞术测定肾炎患者外周血单个核细胞尿激酶型纤溶酶原激活剂受体、IL - 2R和整合蛋白 β2 水平 ,双抗体夹心ELISA法测定血清IL - 2 ,IL - 4和IL - 10水平。结果　肾炎患者外周血单个核细胞尿激酶型纤溶酶原激活剂受体、IL - 2R和整合蛋白 β2 表达水平分别为 (15 .94± 3.2 1) %、(10 .93± 4.2 4) %和 (12 .87± 4.79) % ,均显著高于正常组的 (4.2 1± 1.0 2 ) %、(1.49± 0 .47) %和 (2 .0 8± 1.0 1) % (P <0 .0 1) ;血清IL - 2和IL - 4水平分别为 (2 1.14± 8.2 4) μg/L和 (43.5 6± 10 .2 4) μg/L ,亦均显著高于正常组的 (8.73± 4.0 2 ) μg/L和 (2 3.0 4± 4.78) μg/L(P <0 .0 1) ;IL - 10水平 (8.34± 2 .0 1) μg/L则显著低于正常组的 (12 .0 1± 2 .34 ) μg/L(P <0 .0 1) ;89例肾炎患者外周血单个核细胞尿激酶型纤溶酶原激活剂受体表达水平随病情好转而逐渐恢复正常 ,13例患者激素治疗 2周病情无缓解 ,但外周血单个核细胞尿激酶型纤溶酶原激活剂受体表达水平也逐渐恢复。结论　肾炎患者外周血单个核细胞尿激酶型纤溶酶原激活剂受体表达水平与多种细胞因子有关 ,参与肾小球肾炎的发病 ,在肾小球基底膜?     

肝素对肾炎患者单个核细胞及其尿激酶型纤溶酶原激活剂受体的影响

目的　探讨肝素对肾炎患者尿液单个核细胞分布密度及其尿激酶型纤溶酶原激活剂受体表达的影响。方法　采用流式细胞术方法测定 87例肾炎患者肝素治疗前后尿液单个核细胞分布密度及其该类受体表达水平。结果　肝素治疗前肾炎患者尿液中CD3+、CD14 +和CD16+细胞分布密度显著高于正常组 (P <0 .0 1) ;肝素治疗后尿液CD3+、CD14 +和CD16+细胞分布密度显著减小 ,其中CD14 +和CD16+细胞下降比例较大 ;肝素治疗后尿液单个核细胞该类受体表达水平显著低于治疗前 (P <0 .0 1) ,降至治疗前的 1/4～ 1/3 ;尿液单个核细胞分布密度及其该类受体表达水平与 2 4h尿蛋白定量显著性相关。结论　肝素可显著抑制单个核细胞的免疫活性及其该类受体的表达 ,从而减轻单个核细胞对肾脏的炎性浸润及该受体诱导的肾组织损伤     

尿激酶型纤溶酶原激活剂在小白菊内酯抑制胃癌细胞转移中的作用

目的：探讨小白菊内酯（Par）抑制人胃癌SGC7901细胞转移活性及对尿激酶型纤溶酶原激活剂（uPA）的调控作用。方法：MTT法检测SGC7901细胞增殖能力的变化;倒置显微镜下观察细胞形态;transwell小室法检测细胞迁移和侵袭能力;Western blot法检测细胞uPA蛋白表达;RT-PCR法检测uPA的RNA表达。结果：在100-200μmol/L浓度范围内,Par以浓度和时间依赖方式抑制胃癌细胞增殖,倒置显微镜下见到细胞生长明显受到抑制,细胞的迁移能力和侵袭力均显著下降,穿过人工基底膜的细胞数逐渐减少。随着Par作用时间的延长,SGC7901细胞表达uPA水平逐渐下降。结论：Par可以抑制SGC7901的增殖和转移,uPA在Par降低胃癌细胞转移活性中起重要作用。     

尿激酶型纤溶酶原激活剂在胶质母细胞瘤中表达及意义

目的研究尿激酶型纤溶酶原激活剂(urokinase-typeplasminogenactivator,uPA)在胶质母细胞瘤中的表达及临床意义。方法应用免疫组化LSAB法检测30例胶质母细胞瘤中uPA的表达,结合临床随访,使用Cox回归统计分析。结果uPA染色定位于肿瘤细胞和血管内皮细胞。Cox回归分析显示uPA是影响生存时间的一个独立的预后因子,它与预后存在一定的负相关关系。结论uPA可能是预测胶质母细胞瘤预后的一个独立的预后因子。     

注射用人组织尿激酶型纤溶酶原激活剂质量标准研究

目的:建立注射用人组织尿激酶型纤溶酶原激活剂(HTUPA)的质控方法和质量标准。方法:以纤维蛋白平板法测定HTUPA 的生物学活性,还原型 SDS-PAGE 确定相对分子质量,SDS-PAGE 和反相高效液相色谱测定纯度,毛细管电泳法测定等电点,用胰蛋白酶酶切后分析肽图,其余检测项目按2005年版中国药典三部规定进行。结果:用建立的方法对原液和成品进行了检定,各项指标均符合《人用重组 DNA 制品质量控制技术指导原则》和2005年版中国药典三部的要求。结论:建立的质控方法和质量标准具有保证产品安全、有效、质量可控的特点,可用于注射用人组织尿激酶型纤溶酶原激活剂产品的常规检定。     

重组组织型纤溶酶原激活剂与尿激酶治疗急性ST段抬高型心肌梗死的疗效对比

目的对重组人组织型纤溶酶原激活剂突变体-瑞替普酶(rt-PA)和尿激酶(UK)在急性ST段抬高型心肌梗死患者溶栓疗效的比较。方法选取2011年11月至2013年5月于岳阳市二人民医院心内科就诊的急性ST段抬高型心肌梗死患者63例,随机分为2组,分别给予rt-PA和UK进行溶栓治疗,即分为rt-PA组和UK组,比较2组溶栓再通率、并发症发生、心肌酶学指标、心电图、心功能预后等情况。结果 rt-PA组12 h内溶栓成功率为78.8%,UK组12 h内溶栓成功率为56.7%,前组显著高于后组(P<0.05),左心衰竭、心源性休克并发症的发生率rt-PA组比UK组更小,2组比较差异有统计学意义(P<0.05)。治疗3 h后,ST段回降情况rt-PA组优于UK组,且cTn-T峰值水平前者低于后者(P<0.05),血管再通时间前者短于后者(P<0.05)。治疗30 d后,rt-PA组的心功能恢复情况显著优于UK组(P<0.05)。结论 rt-PA组在溶栓成功率及安全性方面均优于UK组。     

单链尿激酶型纤溶酶原激活剂作用机制与临床应用的研究现状

单链尿激酶型纤溶酶原激活剂(scu-PA)是一种内源性丝氨酸蛋白酶,具有溶栓作用.重组工程化表达的scu-PA已成为一种在临床中广泛应用的溶栓药物,具有溶栓开通率高、全身/颅内出血率低、仅对闭塞性血栓有效、溶栓后再栓率低等特点.除在心肌梗死治疗中得到广泛应用外,其在急性缺血性脑卒中、急性肺栓塞、下肢深静脉血栓形成等血栓...     

喉咽癌组织中胰岛素生长因子1受体与尿激酶型纤溶酶原激活剂的表达及意义

目的探讨胰岛素生长因子1受体（IGF-1R）与尿激酶型纤溶酶原激活剂（u—PA）在喉咽癌中的表达及其相互关系。方法采用免疫组织化学链霉菌抗生物素蛋白-过氧化物酶连结法（SP法）,分别检测55例喉咽癌（咽喉癌组）、42例癌旁喉咽黏膜（癌旁组）及40例会厌囊肿（对照组）中IGF-1R蛋白和u-PA蛋白的表达情况。结果①IGF-1R与尿激酶型纤溶酶原激活剂u—PA在喉咽癌组、癌旁组及对照组中的阳性表达率及强阳性率依次降低（81．82％,52．38％,17．50％）,3组间两两比较,差异均有统计学意义（P〈0．01）;②IGF-1R与U-PA在喉咽癌有淋巴结转移组的强阳性表达率分别为82．61％和78．26％,与无淋巴结转移组中22．22％和44．44％相比,差异有统计学意义（P〈0．01或P〈0．05）。③IGF-1R与u-PA在喉咽癌组织中的表达有明显的相关性（P〈0．01）。结论①IGF-1R和u—PA在喉咽癌组织中高表达可能与喉咽癌的发生发展有关。②检测喉咽癌组织中IGF-1R和u—PA的表达,可能有助于估计喉咽癌淋巴结转移。③IGF-1R与u—PA可能在喉咽癌发生、发展以及淋巴结转移中起了协同作用。     

靶向尿激酶型纤溶酶原激活物受体的抗肿瘤药物的研究进展

尿激酶型纤溶酶原激活物受体（u PAR）是一种糖基磷脂酰肌醇锚定的膜蛋白,已被发现在多种癌细胞中过表达,这一特性使得uPAR成为治疗癌症的一个理想靶标。目前,多种以u PAR为靶点的抗癌药物已被开发,主要包括多肽、单克隆抗体、配体靶向毒素。此外,u PAR作为靶点也已在纳米药物递送系统和光热疗法（PTT）/光动力疗法（PDT）中得到应用。因此就靶向uPAR的抗肿瘤药物的研究进展做一综述。     

Chemotactic effect of urokinase-type plasminogen activator on mouse spermatozoa in vitro

The aim of this study is to investigate the chemotactic effect of urokinase-type plasminogen activator (uPA) on mouse spermatozoa. Capillary assays were applied to study the chemotactic activity of ascending and descending gradients of uPA. Firstly, the chemotactic effect of an ascending gradient of uPA on mouse spermatozoa was observed by counting the number of spermatozoa that migrated into the capillary after incubation with uPA for 5, 10, 20, and 30 min, respectively, compared with that after incubation with F10. Twenty minutes was a suitable incubation time to obtain a plateau of sperm accumulation. Meanwhile, to confirm the specific effect of uPA on mouse sperm chemotaxis, uPA inhibitor (PAI-1) and anti-uPAR rabbit IgG were added to the test solution containing 20 U/mL uPA, respectively. To exclude the possibility that PAI-1 and anti-uPAR rabbit IgG may affect sperm accumulation nonspecifically, PAI-1 and anti-uPAR rabbit IgG were added to F10, respectively. It was found that the chemotactic effect of uPA was neutralized completely by PAI-1 and anti-uPAR rabbit IgG. PAI-1 and anti-uPAR rabbit IgG had no neutralizing effect on the sperm chemotactic effect. Lastly, the sperm chemotaxis response to a descending gradient of uPA was also observed. Taken together, the results suggest that uPA can induce sperm chemotaxis in vitro by binding to its receptor on the sperm membrane and may act as a chemoattractant in precontacting sperm-egg communication thereby increasing the chance encounter of spermatozoa and eggs.     

Small molecule inhibitors of urokinase-type plasminogen activator

The urokinase-type plasminogen activator (uPA) protein is a multifunctional protein involved in a myriad of biological activities including extracellular matrix degradation and cell invasion. Active uPA is a 411 amino acid protein consisting of 3 domains, each of which confers a particular biological function to the overall protein. The amino terminal domain or growth factor domain (GFD), comprised of amino acid residues 1 – 48, is involved in uPA interaction with its cell surface receptor, urokinase-type plasminogen activator receptor (UPAR). The interaction of uPA with UPAR promotes, in part, cell adhesion, migration and invasion. A second domain is the kringle domain, comprising amino acid residues 49 – 135. Initially thought to bind heparin, the kringle domain has more recently been shown to possess antiangiogenic activity. A third domain comprising amino acid residues 159 – 411, the serine protease domain, is involved in the proteolytic activation of plasminogen to plasmin. The production of plasmin by uPA begins a cascade of events manifested by extracellular matrix degradation. The recent patent literature describes small molecule compounds, which inhibit the interaction of uPA with UPAR, inhibit the proteolytic activity of the uPA serine protease domain and inhibit the interaction of uPA with its natural inhibitor, plasminogen activator inhibitor-1 (PAI-1). Small peptides encompassing residues 19 – 31 of the GFD have been developed which exhibit potent inhibition of the uPA–UPAR interaction and show efficacy in tumour-bearing animal models. Small molecules have been disclosed by Corvas, which are reported to be inhibitors of PAI-1. Finally, two approaches toward the development of inhibitors of the uPA serine protease domain have been described in the recent patent literature. The first approach describes non-covalent peptidederived inhibitors discovered by phage display techniques, which bind in the substrate-binding groove of the uPA active site. An alternative approach describes non-covalent small molecule inhibitors, which bind in the enzyme active site in a slightly different binding mode than the peptide-derived inhibitors. These small molecule non-peptide analogues inhibit the uPA proteolytic activity quite effectively and are reported to possess excellent enzyme selectivity and highly improved oral activity. The clinical utility of small molecule uPA enzyme inhibitor analogues awaits the results of a preliminary clinical evaluation of compounds described by Wilex.     

Cancer treatment with inhibitors of urokinase-type plasminogen activator and plasmin

The urokinase-type plasminogen activator-plasmin system plays an important role in many normal physiological processes including clot lysis, wound healing, embryogenesis and tissue remodelling. It is also involved in the pathogenesis of human malignancy through its ability to mediate tumour cell growth, invasion and metastatic dissemination. Interfering with this system is an appealing approach for experimental therapy of malignancy for several reasons. This concept is supported by a wealth of preclinical data. Evidence exists suggesting a role for this system in several major human tumour types. Preliminary evidence suggests that agents which block this pathway are effective in therapeutic doses that are already defined and relatively non-toxic. This form of treatment is not likely to carry cross-resistance with other types of cancer therapy and should be applicable to both localised and advanced tumours. Since heterogeneity in responsiveness among various tumour types is expected, clinical effects in given tumours would provide a basis for interpreting mechanisms of tumour progression in vivo and for future development of drugs with improved efficacy. Inhibition of the urokinase-type plasminogen activator-plasmin system remains a promising, but largely untested, area of experimental cancer therapeutics.     

Antischizophrenic effect of dopamine D3 receptor antagonist Y-QA31

OBJECTIVCE Schizophrenia is a chronic,debilitating psychotic mental disorder.Many studies suggest that dopamine D3 receptor(DAD3R) plays an important role in etioloy of schizophrenia,and selective DAD3R antagonists may have effects to improve the symptoms of schizophrenia.More important,the DAD3R antagonists result in less or no extrapyramidal side effect(EPS).Therefore,the present study is aimed to evaluate the pharmacological effects of Y-QA31 which was found to be a highly selective DAD3R antagonist,hoping to develop new potential APDs.METHODS We use the animals models,including methamphetamine(MA) or dizocilpine(MK-801) induced hyperactivity,apomorphine-induced climbing,the disruption of prepulse Inhibition elicited by MA,conditioned avoidance responding and apomorphine-induced hypothermia to evaluate the effects of Y-QA31 on the treatment of schizophrenia.Meanwhile,the induction of catalepsy of Y-QA31 was also observed in mice.RESULTS Y-QA31(20-40 mg·kg-1,po) effectively inhibited conditioned avoidance responding in mice,the hyperactivity induced by MA or MK-801,and apomorphine-induced hypothermia in mice by dose-dependence.Y-QA31(5 mg·kg-1,po) could also inhibit the reduction of prepulse Inhibition elicited by MA in mice.Morerover,Y-QA31(80 mg·kg-1,po) could also inhibit apomorphine-induced climbing in mice.Y-QA31 did not induce catalepsy when its dose increased to 200 mg·kg-1 po,suggesting the EPS of Y-QA31 was very low.CONCLUSIONThe above results indicated Y-QA31 exhibited the therapeutic effects on positive symptoms of schizophrenia with low EPS,and further suggested that DAD3R is the potential target in the treatment of schizophrenia.     

Increased vulnerability to cocaine addiction in mice lacking dopamine D3 receptor

Neuroimaging studies using positron emission tomography suggest that reduced dopamine(DA) D2 receptor(D2R) availability in the striatum is associated with increased vulnerability to drug addiction in humans and experimental animals.However,the role of D3R in the neurobiology of addiction remains unclear.Here we report that D3R-knockout(D3-/-) mice display enhanced cocaine(and sucrose) taking observed during the acquisition and maintenance of cocaine self-administration and enhanced motivation for cocaine(and sucrose) seeking observed during progressive-ratio cocaine self-administration and extinction from cocaine self-administration.This increased vulnerability to cocaine was accompanied by decreased DA response to cocaine secondary to increased basal levels of extracellular DA in the nucleus accumbens,suggesting that enhanced cocaine-taking and cocaine-seeking behavior could be a compensatory response to decreased cocaine reward in D3-/-mice.In addition,D3-/-mice also displayed up-regulation of DA transporter in the striatum,suggesting that a neuroadaptative change occurred D3-/-mice to restore elevated basal levels of extracellular DA.These findings,for the first time,suggest that deletion of D3R increases vulnerability to cocaine-taking and cocaine-seeking behavior.Thus,reduced D3R availability in the brain constitutes an important risk factor for the development of cocaine addiction.     

Antipsychotic regulation of dopamine D1, D2 and D3 receptor mRNA

A range of antipsychotic drugs, both “typical” and “atypical”, was administered to rats over a time course and at several different dosages. The mRNA levels of dopamine D₁ , D₂ and D₃ receptor were measured in either whole brain or dissected brain regions. D₃ receptor mRNA was up-regulated in whole brain by clozapine (10 and 30 but not 3 mg/kg/day), sulphide (50 and 100 but not 20 mg/kg/day), haloperidol (3 but not 1 or 0.3 mg/kg/day), flupenthixol (3 but not 1 or 0.3 mg/kg/day), pimozide (4.5 but not 1.5 or 0.5 mg/kg/day) and loxapine (1.2 and 4 mg/kg/day but not 0.4 mg/kg/day). Sulphide (100 mg/kg/day), clozapine (30 mg/kg/ day) and haloperidol (3 mg/kg/day) all up-regulated the D₃ receptor mRNA in nucleus accumbens and olfactory tubercles but not striatum. D₁ and D₂ receptor mRNA was up-regulated in whole brain by haloperidol and loxapine only, and in the case of haloperidol this was localized to striatum and prefrontal cortex. Haloperidol, clozapine and sulphide all down-regulated D₁ mRNA in hippocampus and additionally haloperidol and sulpiride down-regulated it in the cerebellum. This work shows that all the drugs tested upregulated D₃ receptor, but effects on D₁ and D₂ receptors were less general.