糖尿病性和非糖尿病性脑梗死患者血浆中纤溶酶原激活物及其抑制剂活性的动态观察

目的 :观察糖尿病性脑梗死 (diabeticischemicstroke ,DIS)和非糖尿病性脑梗死 (non diabeticischemicstroke ,NDIS)患者血浆中纤溶酶原激活物 (plasminogenactivator ,PA)及纤溶酶原激活物抑制剂 (plasminogenactivatorinhibitor ,PAI)的动态变化情况。方法 :应用底物发色法测定血浆中PA和PAI活性 ,以观察DIS和NDIS患者血浆PA和PAI活性的动态变化。结果 :NDIS患者血浆中PA活性在4～ 2 1d较非脑梗死患者升高 ;DIS患者的PA活性在 7h～ 2 1d较NDIS患者为低 ,但PAI活性在各组间无明显差异。结论 :DIS患者血浆中PA活性较NDIS患者降低 ,提示其存在纤溶系统激活的紊乱 ,并可能与DIS症状加重有关。     

急性脑出血患者血浆及脑脊液组织型纤溶酶原激活物及其抑制物含量的观察

目的　研究急性脑出血患者血浆及脑脊液组织型纤溶酶原激活物 (t PA)及其抑制物 (PAI 1)含量的变化及其临床意义。方法　采用双抗体夹心固相酶联免疫吸附法 (ELISA)检测 34例急性脑出血患者血浆和其中 2 1例患者脑脊液t PA及PAI 1抗原含量 ,并与正常对照组的血浆和脑脊液t PA、PAI 1含量进行比较。结果　急性脑出血组患者血浆及脑脊液t PA、PAI 1含量均显著高于对照组 (P <0 0 1,P <0 0 5 ) ;脑脊液中t PA、PAI 1的含量分别与血浆中t PA、PAI 1的含量呈正相关 (均P <0 0 5 )。结论　急性脑出血患者纤溶活性明显升高 ;t PA及PAI 1含量是反映体内纤溶活性的两个重要指标     

急性脑血栓形成患者脑脊液组织型纤溶酶原激活物及其抑制物含量测定的意义

目的 本文通过观察急性动脉硬化性脑血栓形成患者脑脊液组织型纤溶酶原激活物(t—PA)及其抑制物(PAI—1)的抗原含量，以探讨急性脑血栓形成患者的纤溶活性及其临床意义。方法 采用双抗体夹心固相酶联免疫吸附法(ELISA)检测31例患者脑脊液t—PA及PAI—1抗原含量，与20名对照组脑脊液进行比较。结果 急性脑血栓形成组脑脊液t—PA、PAI—1含量均显著高于对照组。结论 说明急性脑血栓形成患者纤溶活性明显下降，t—PA及PAI—1参与了脑血栓形成之病理过程；t—PA及PAI—1抗原含量是反映体内纤溶活性的2个重要指标。     

急性脑梗死患者凝血纤溶机制的探讨

目的 :初步探讨急性脑梗死患者血浆及脑脊液凝血纤溶变化机制 ,为临床提供诊治依据。方法 :急性脑梗死患者 35例 (采集血浆 ) ,其中 31例同时采集脑脊液 ,测定脑脊液及血浆部分凝血纤溶指标 ,并与正常对照组进行比较。结果 :急性脑梗死组脑脊液及血浆组织型纤溶酶原激活物 (t- PA)、 D-二聚体 (D- D)含量明显高于对照组 (P<0 .0 1) ;而纤溶酶原 (PL G)活性明显下降 (P<0 .0 1)。结论 :脑梗死急性期存在明显的高凝状态和继发性纤溶活性增高     

水蛭提取液对培养的大鼠脑皮质微血管内皮细胞分泌组织型纤溶酶原激活物和纤溶酶原激活剂抑制物1的影响

目的探讨水蛭提取液(HEL)对培养的大鼠脑皮质微血管内皮细胞分泌组织型纤溶酶原激活物(tPA)、纤溶酶原激活剂抑制物1(PAI-1)的影响。方法建立大鼠大脑皮质微血管内皮细胞培养实验模型。MTT法筛选HEL的有效浓度。检测培养上清液的tPA、PAI-1含量与活性变化,RT-PCR检测经HEL治疗组与生理盐水对照组处理后的微血管内皮细胞tPA与PAI-1的表达,免疫组化检测两组微血管内皮细胞tPA的表达。结果 HEL在一定浓度范围内(0.25～1mg/μl)可促进微血管内皮细胞的生长,有剂量依赖关系(P<0.05)。HEL治疗组较生理盐水对照组能促进培养的大鼠脑皮质微血管内皮细胞分泌tPA,同时提高其活性,促进tPA mRNA的表达及tPA免疫活性表达,且呈剂量依赖性表达增强(P<0.01)。结论 HEL在体外能激活内源性纤溶系统。     

急性脑梗死患者血浆纤溶酶原激活物及其抑制剂-1水平的动态变化及其与梗死面积的关系

目的 探讨急性脑梗死患者血浆组织型纤溶酶原激活物(t-PA)及其抑制剂-1(PAI-1)水平的动态变化及其与梗死面积的关系.方法 急性脑梗死患者100例,其中大面积脑梗死22例、小面积脑梗死36例、腔隙性脑梗死42例,采用发色底物显色法检测脑梗死患者病后24 h、2 d、14 d、21 d的血浆t-PA、PAI-1水平,与正常对照组比较;并比较不同面积脑梗死患者血桨t-PA、PAI-1水平.结果 与正常对照组比较,急性脑梗死患者病后24 h、2 d、14 d血浆t-PA水平显著降低,血浆PAI-1水平明显升高(均P＜0.01);病后21 d两者与正常对照组差异无统计学意义(均P＞0.05);大面积脑梗死患者t-PA水平明显低于小面积和腔隙性脑梗死患者,小面积脑梗死患者又明显低于腔隙性脑梗死患者(均P＜0.01);不同面积脑梗死组之间PAI-1水平未见明显差异(均P＞0.05).结论 脑梗死患者急性期血浆t-PA水平降低及PAI水平升高;脑梗死面积越大的患者血浆t-PA水平降低程度越明显,而血浆PAI-1水平与梗死面积无关.     

脑出血患者尿激酶型纤溶酶原激活物及其受体的变化

目的探讨脑出血患者血浆尿激酶型纤溶酶原激活物(uPA)及其受体(uPAR)的变化及临床意义。方法对64例脑出血患者和30例健康对照组应用酶联免疫吸附双抗体夹心法(ELISA法)定量测定血浆uPA和u-PAR的水平。结果脑出血组uPA含量(2866±788ng/L)和uPAR含量(3645±322ng/L)明显高于正常对照组的uPA含量(1075±244ng/L)和uPAR含量(877±216nP<0.01)。随脑出血患者病情严重程度的加重,血浆uPA和u-PAR的含量增高越明显。结论血浆uPA和uPAR的增高参与了脑出血时继发性损害,与脑出血患者的病情密切相关。     

自发性蛛网膜下腔出血后血液及脑脊液纤溶活性的研究

目的探讨自发性蛛网膜下腔出血(SAH)后血及脑脊液(CSF)中纤溶活性的变化规律及正常CSF中的纤溶状态。方法组织型纤溶酶原激活物的活性(tPA:A)及纤溶酶原抑制物的活性 (PAI:A)测定采用发色底物法,纤溶酶原抑制物-1(PAI-1)及D-二聚体(D-D)定量测定采用酶联免疫吸附试验(ELISA)双抗体夹心法,血及CSF标本在SAH后0-3 d(急性期),4-9 d(再出血高峰期),及 14-21 d(吸收期)留取三次;正常对照组留取一次。结果 SAH患者血液中,急性期tPA:A显著低于对照组,并随病程延长显著升高,至14-21 d达正常水平,与对照组差异无显著意义;急性期PAI:A 及D-D水平显著高于对照组,并随病程延长而显著降低,至14-21 d降至正常水平,与对照组差异无显著意义;各期PAI-1含量与对照组差异无显著意义。对照绀CSF中,测不到tPA:A及PAI:A及 PAI-1,D-D测得值很小,为(0．28±0．36)mg/L。 SAH患者CSF中,各期测不到tPA:A及PAI:A,但 PAI-1含量急性期显著升高并随病程延长而显著降低,D-D急性期显著升高并随病程延长而显著降低,至14-21 d 与对照组无显著性差异。结论 SAH后血中不存在纤深活性亢进。对照组CSF中不含有纤溶酶系。SAH患者CSF中纤溶活性急性期升高而后降低,于14-21 d恢复到正常水平。再出血高峰期血及CSF中反映纤溶活性的指标变化均显著低于急性期,提示再出血与血及CSF中纤溶活性无关。故SAH后不宜长期大剂量应用抗纤溶药物来预防再出血。     

脑梗死患者的胰岛素抵抗与血浆组织型纤溶酶原激活物抑制物-1活性相关性的研究

目的　探讨动脉硬化性脑梗死 (ACI)患者急性期和恢复期的胰岛素抵抗 (IR)与血浆组织型纤溶酶原激活物 (t PA)抑制物 1(PAI 1)活性的关系。方法　分别测定 91例ACI患者急性期和恢复期血糖、胰岛素水平和血浆t PA和PAI 1的活性。血糖与胰岛素乘积之倒数的自然对数作为胰岛素敏感性指数 (ISI) ,并与4 0名健康同龄人对照。结果　ACI患者急性期和恢复期的血糖、胰岛素水平及PAI 1活性显著高于对照组(P <0 0 1～ 0 0 5 ) ,ISI和t PA活性显著低于对照组 (P <0 0 1～ 0 0 5 ) ;PAI 1活性与胰岛素水平显著正相关(r=0 .6 7,r=0 .6 6 ,均P <0 0 1) ,与ISI显著负相关 (r=- 0 .85 ,r=- 0 .6 6 ,均P <0 0 1) ;t PA活性与这些参数不相关 ;脑梗死体积与ISI负相关 (r=- 0 .19,P <0 .0 5 ) ,与PAI 1活性正相关 (r=0 .5 6 ,P <0 .0 5 ) ;体重指数与ISI负相关 (r=- 0 .4 9,P <0 .0 5 ) ,与PAI 1活性正相关 (r=0 .5 3,P <0 .0 5 )。结论　IR提高ACI患者的血浆PAI 1活性 ,IR的个体处于脑血栓形成的危险之中     

急性脑梗死患者尿激酶型纤溶酶原激活物及受体的变化

目的探讨急性脑梗死患者血浆尿激酶型纤溶酶原激活物(uPA)及受体(uPAR)的变化和临床意义。方法对66例急性脑梗死患者和30例健康者应用酶联免疫吸附双抗体夹心法(ELISA法)定量测定血浆uPA和uPAR的水平。结果与正常对照组相比,急性脑梗死组uPA含量(4756±632)ng/L,uPAR含量(3102±256)ng/L,明显高于正常对照组的uPA(1075±244)ng/L和uPAR(877±216)ng/L,差异有显著性(P<0.01)。随急性脑梗死患者病情严重程度的加重,血浆uPA和uPAR含量增高越明显。结论急性脑梗死患者血浆uPA和uPAR的水平明显升高,且与急性脑梗死患者的病情密切相关。     

Reductions in mRNA of the neuroprotective agent, neuroserpin, after cerebral ischemia/reperfusion in diabetic rats

The aim of this study is to investigate disturbances in fibrinolytic components in diabetic rats with middle cerebral artery occlusion (MCAO). Comparison of cerebral injury at 23 h after reperfusion indicated that infarction volumes were increased in diabetic rats as compared with normal rats. Cerebral ischemia/reperfusion in normal and diabetic rats was accompanied by increased expression of tissue plasminogen activator (t-PA), urokinase plasminogen activator (u-PA), plasminogen activator inhibitor 1 (PAI-1) and neuroserpin (NSP) mRNA after reperfusion. Most importantly, the expression of NSP mRNA, but not t-PA, u-PA and PAI-1 mRNA, was reduced to undetectable levels at 11 and 23 h after reperfusion in diabetic rats as compared with normal rats. Although activity of PA (t-PA and u-PA) and the ratio of PA/PAI were increased at 5 h after reperfusion in both ischemic brains of diabetic and normal rats, the levels in diabetic rats were lower than that in normal rats. We speculate that the exacerbation of ischemic injury in diabetic rats may be related to the reduction of fibrinolytic component and the neuroprotective role of NSP. Further study of the efficacy of NSP in the pathogenesis and treatment of cerebral ischemia may provide novel insights.     

Rt-PA causes a significant increase in endogenous u-PA during experimental focal cerebral ischemia

The aim of this study was to investigate the effects of different doses of exogenous recombinant human tissue plasminogen activator (rt-PA) on the endogenous cerebral plasminogen-plasmin system in focal ischemia in rats. Ischemia was induced using the suture model. Each group of rats (n = 6) received either treatment (0.9, 9 or 18 mg rt-PA/kg body weight) or saline (control group) at the end of ischemia; a sham-operated group was added. The activity of the plasminogen activators was measured by casein-dependent plasminogen zymography. In the cortex urokinase (u-PA) rose from sham (no ischemia), 91 +/- 7% to ischemia, 176 +/- 10% (P < 0.005). Increasing rt-PA doses led to further significant (P < 0.001) cortical u-PA activation which was maximal at 18 mg: 249 +/- 13%. An extreme increase in the u-PA activity was observed in the basal ganglia to 1019 +/- 22% (P < 0.001). This increase was further aggravated by higher rt-PA doses (18 mg, 1236 +/- 15%; P < 0.001). The t-PA level did not change I3R24 during (3 h ischemia followed by reperfusion for 24 h); however, during low and moderate doses of rt-PA, endogenous t-PA was reduced. In conclusion, while ischemia leads to a significant increase in u-PA, mainly in the basal ganglia, t-PA is not altered. Increasing doses of rt-PA lead to a further elevation of u-PA. Thus, u-PA seems to play a major role in the endogenous plasminogen activator system following focal cerebral ischemia.     

Plasminogen activation in experimental permanent focal cerebral ischemia

BACKGROUND: Previous experimental work using in situ zymography has shown very early increased plasminogen activation in ischemic regions after 3 h of ischemia with and without reperfusion. The objective of the present study was to evaluate the time course and extent of plasminogen activation in long-term permanent focal cerebral ischemia. MATERIAL AND METHODS: The middle cerebral artery in male Fisher rats was irreversibly occluded by electrocoagulation. Duration of ischemia was 48, 72, and 168 h. Occlusion was controlled in vivo by MRI at day 2. Plasminogen activation was detected by in situ zymography of 10 microm cryosections with an overlay containing plasminogen and the plasmin substrate caseine. Areas of plasminogen activation were compared to structural lesions (immunohistochemical loss of microtubule-associated protein 2; MAP 2). RESULTS: Compared to controls, increased plasminogen activation was observed in the basal ganglia and the cortex of the ischemic hemisphere after 48, 72, and 168 h (affected area of basal ganglia: 44.5+/-21.9, 70.1+/-2.3 and 66.6+/-2.8%, respectively; affected area of cortex: 63.4+/-9.8, 67.7+/-0.7 and 64.0+/-3.7%, respectively). The duration of ischemia had no significant influence on the extent of plasminogen activation. Areas of increased plasminogen activation significantly overlapped with and exceeded areas of MAP 2 loss (P<0.005). DISCUSSION: Permanent focal cerebral ischemia leads to increased plasminogen activation in ischemic regions. This plasminogen activation remains elevated at persistent levels over days. It may contribute to extracellular matrix (ECM) disruption, secondary hemorrhage, and brain edema in subacute stages of ischemic stroke.     

缺血后处理对糖尿病大鼠局灶性脑缺血再灌注损伤线粒体的影响

目的观察缺血后处理(I-Post)对糖尿病大鼠局灶性脑缺血再灌注损伤线粒体超微结构和功能的影响,探讨I-Post诱导的脑保护的可能机制。方法采用链脲佐菌(STZ)腹腔注射建立糖尿病大鼠模型,在此基础上通过线栓法建立大鼠大脑中动脉阻塞/再灌注模型。SD糖尿病大鼠随机分为4组(n=10),空白对照组、假手术组、缺血再灌注组(I/R组)、缺血后处理组(I-Post组)。于缺血90min再灌注6h后电镜下观察线粒体超微结构、测定缺血侧脑组织线粒体中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)、Na+/K+-ATPase和Ca2+-ATPase活性。结果缺血后处理能明显减轻I/R引起的线粒体超微结构的损伤,提高线粒体SOD、GSH-Px、Na+/K+-ATPase和Ca2+-ATPase的活性(P<0.05或0.0),降低MDA的含量(P<0.05)。结论线粒体可能在I-Post诱导的脑保护中起关键性作用,I-Post诱导的脑保护机制可能与SOD、Na+/K+-ATP酶、Ca2+-ATP酶和GSH-Px活性增加有关。     

肢体缺血后处理对糖尿病大鼠脑缺血再灌注损伤线粒体结构和功能的影响

目的探讨肢体缺血后处理(I-PostC)诱导的远隔器官I-PostC(RPostC)对糖尿病大鼠局灶性脑缺血再灌注(I/R)损伤线粒体结构和功能的影响。方法链脲佐菌素空腹腹腔注射制作糖尿病大鼠模型,线栓法闭塞大脑中动脉(MCAO)制作局灶性I/R大鼠模型。造模成功的40只雄性SD大鼠分为4组:空白对照组、假手术组、I/R组,RPostC组,每组10只。I/R6h后取脑组织行HE染色观察,测定线粒体丙二醛(MDA)含量、超氧化物歧化酶(SOD)、Na+/K+-ATP酶、Ca2+-ATP酶和谷胱甘肽过氧化物酶(GSH-Px)活性,观察线粒体超微结构的改变。结果I/R组线粒体MDA含量[(4.99±1.25)nmol/mgprot]较空白对照组和假手术组明显升高(均P<0.01),SOD[(72.52±13.07)U/mg]、Na+/K+-ATP酶[(3.17±0.34)μmolPi/(mg.h)]、Ca2+-ATP酶[(1.56±0.23)μmolPi/(mg.h)]和GSH-Px活性[(22.66±5.29)U/mg)]明显降低(均P<0.01);与I/R组比较,RPostC组MDA含量[(3.58±0.91)...     

槲皮素对糖尿病脑梗死大鼠脑纤溶激活系统基因表达的研究

目的 观察槲皮素对糖尿病脑梗死大鼠脑组织中纤溶酶原激活物(t-PA)及其抑制剂 mRNA表达的影响.方法 Wistar大鼠50只,随机分为正常假手术组、糖尿病假手术组、糖尿病脑梗死组、低剂量与高剂量槲皮素预处理组5组.后4组大鼠制作糖尿病模型.造模后低剂量与高剂量槲皮素预处理组分别按体质量予槲皮素50 mg/(kg·d)、100 mg/(kg·d)灌胃15 d,并与糖尿病脑梗死组行自体血栓脑梗死处理.以半定量反转录-聚合酶链式反应法(RT-PCR)检测各组动物脑组织中组织型纤溶酶原激活物(t-PA) mRNA、尿激酶型纤溶酶原激活物(u-PA) mRNA、1型纤溶酶原激活物抑制剂(PAI-1) mRNA、神经丝氨酸蛋白酶抑制剂(NSP) mRNA的表达并进行分析比较.结果 与糖尿病脑梗死组t-PA mRNA、PAI-1 mRNA相对表达量(分别为0.27±0.03、0.68±0.06)比较,低剂量、高剂量槲皮素预处理组t-PA mRNA表达(分别为0.32±0.05、0.44±0.10)升高,PAI-1 mRNA表达(0.53±0.15、0.32±0.07)降低(均P<0.01),高剂量、低剂量槲皮素预处理组、糖尿病脑梗死组之间u-PA mRNA、NSP mRNA表达无统计学差异.结论 槲皮素可能通过促进t-PA mRNA表达、抑制PAI-1 mRNA表达改善糖尿病脑梗死大鼠纤溶功能.槲皮素预处理对糖尿病脑梗死大鼠u-PA mRNA、NSP mRNA表达无明显影响.     

Plasminogen activation in focal cerebral ischemia and reperfusion.

In focal cerebral ischemia the plasminogen-plasmin system plays a role in the fibrinolysis of vessel-occluding clots and also in the proteolysis of extracellular matrix components, which potentially contributes to brain edema and bleeding complications. The authors investigated the plasminogen activation after middle cerebral artery occlusion with and without reperfusion (reperfusion intervals 9 and 24 hours) in rats by histologic zymography and compared areas of increased plasminogen activation to areas of structural injury, which were detected immunohistochemically. After 3 hours of ischemia, increased plasminogen activation was observed in the ischemic hemisphere. The affected area measured 5.2%+/-8.5% and 19.4%+/-30.1% of the total basal ganglia and cortex area, respectively. Reperfusion for 9 hours after 3 hours of ischemia led to a significant expansion of plasminogen activation in the basal ganglia (68.8%+/-42.2%, P < 0.05) but not in the cortex (43.0%+/-34.6%, P = 0.394). In the basal ganglia, areas of increased plasminogen activation were related to areas of structural injury (r = 0.873, P < 0.001). No such correlation was found in the cortex (r = 0.299, P = 0.228). In this study, increased plasminogen activation was demonstrated early in focal cerebral ischemia. This activation may promote early secondary edema formation and also secondary hemorrhage after ischemic stroke.     

流式细胞术检测局灶性脑缺血再灌注bcl-xl、bax蛋白表达

目的 探讨应用流式细胞术检测bcl-xl和bax在局灶性脑缺血再灌注中的表达及作用。方法 45只健康雄性Wistar大鼠随机分为正常对照组、假手术组。缺血3 h组和缺血3 h再灌注组;缺血再灌注组又分为缺血再灌注3h、6h、12h、24h、48h和72h等6个亚组。采用线栓法复制大脑中动脉脑缺血再灌注模型,应用流式细胞术检测bcl-xl和bax蛋白表达。结果 缺血3 h再灌注各亚组,随再灌注时间的延长,半影区bax表达逐渐增强,于再灌注24 h和48 h达高峰(均P<0.01);bcl-xl表达在早期增强后随即呈下降趋势,bcl-xl/bax比值呈动态变化。再灌注中心区,在缺血再灌注早期bcl-xl和bax表达均增强(P<0.05或P<0.01),随着再灌注时间的延长,二者的表达及bcl-xl/bax比值逐渐减弱。结论 bcl-xl表达增强有利于促进神经细胞存活,而bcl-xl与bax比值的转变则促进神经细胞凋亡。     

肢体缺血预处理对脑缺血再灌注损伤大鼠自噬的影响

目的探讨肢体缺血预处理对脑缺血再灌注损伤大鼠自噬的影响。方法将60只Wistar大鼠随机分为假手术组(Sham组)、缺血再灌注组(I/R组)、肢体缺血预处理组(LIPC组)、3-甲基嘌呤组(3-MA组),每组15只。制作脑缺血再灌注、肢体缺血预处理及3-MA干预大鼠模型,在脑缺血2 h再灌注24 h后进行神经功能缺陷评分和脑梗死体积测定,HE染色观察细胞形态学改变,Western Bloting法检测自噬相关蛋白Beclin-1、Cathepsin B的表达。结果与I/R组比较,LIPC组神经功能缺陷评分降低(P<0.05),脑梗体积明显减小(P<0.05),细胞损伤、坏死减轻(P<0.05),Beclin-1、Cathepsin B的蛋白表达明显减弱(P<0.05)。结论 LIPC对缺血再灌注损伤大脑具有保护作用,其机制可能与减弱自噬水平有关。     

老年脑血栓患者血浆中t－PA、PAI的变化

本文采用发色底物显色法对３９例脑血栓患者血浆中组织型纤溶酶原激活物（ｔ－ＰＡ）和纤溶酶原激活物抑制物（ＰＡＩ）进行检测。结果ｔ－ＰＡ活性下降，ＰＡＩ活性升高，与健康对照组相比有显著差异性（Ｐ＜０．０１），表明脑血栓患者纤溶活性降低，是产生血栓的因素之一。本栓测结果提示，测定ｔ－ＰＡ、ＰＡＩ有助于脑血栓的诊断。