血小板活化与动脉粥样硬化病变相关性的研究进展

随着当今社会心脑血管病发病率的不断攀升，心脑血管疾病的共同病理基础——动脉粥样硬化（atherosclerosis，AS）引起广大医务工作者和医学科研工作者的高度关注。AS是一个与内皮细胞损伤、脂质紊乱、血流动力学异常有关的、极其复杂的慢性炎症过程。     

老年2型糖尿病脂联素与血小板活化和动脉粥样硬化相关性研究

50例2型糖尿病（T2DM）颈总动脉内膜中层厚度（IMT）、血小板活化和胰岛素抵抗指（HOMA—IR）高于对照组，脂联素与血小板活化、颈总动脉IMT和HOMA—IR呈负相关。低脂联素血症与IR和动脉粥样硬化相关，血小板活化（CD62P、CD41＋CD11b）是影响T2DM脂联素水平危险因素之一。     

下肢深静脉血栓形成患者血小板活化状态的变化及意义

为探讨下肢深静脉血栓形成(DVT)思考血小板活化状态的变化及其临床意义，用流式细跑术(PCM)，以单克隆抗体为探针，对35例下肢DVT患者(DVT组)及3l例健康人(对照组)血小板活化标记物溶酶体颗粒糖蛋白(CD63)、血小板表面选择素(CD62p)及凝血酶敏感蛋白(TSD)进行了检测。结果显示，DVT组3种血小板活化标志物阳性表达率均高于对照组(P均＜0．001)，溶栓治疗后3种血小板活化标志物的阳性表达率均呈降低趋势。提示DVT患者体内血小板活化亢进；FCM可作为活化血小板的良好检测手段。     

阻塞性睡眠呼吸暂停低通气综合征患者血小板活化和纤溶活性的研究

目的　探讨血小板活化、凝血激活和继发纤溶亢进在阻塞性睡眠呼吸暂停低通气综合征 (OSAHS)发生发展中的作用及经鼻持续气道正压通气 (nCPAP)对其影响。方法　选择经多导睡眠图 (PSG)确诊的OSAHS患者 5 8例为实验组 ,根据睡眠呼吸暂停低通气指数 (AHI)、最低血氧饱和度(SaO2 min)将OSAHS患者分轻、中、重组 ,并设正常对照组 2 0例 ,11例重度OSAHS患者接受nCPAP治疗为治疗组 ,用酶联免疫双抗体夹心法检测各组血浆α 颗粒膜蛋白 (GMP 14 0 )、血小板膜糖蛋白Ⅱb/Ⅲa(GPⅡb/Ⅲa)和D 二聚体 ,比较各实验组与对照组 ,治疗组治疗前后的各项指标的差异。　结果(1)中、重度OSAHS患者组血浆GMP 14 0、GPⅡb/Ⅲa和D 二聚体均显著高于对照组 (P <0 .0 5 ) ,nCPAP治疗后比治疗前明显下降 (P <0 .0 0 1) ;(2 )GMP 14 0、GPⅡb/Ⅲa和D 二聚体与AHI呈正相关 ,与最低SaO2 呈负相关 (P <0 .0 0 1)。结论　中、重度OSAHS患者存在血小板活化、凝血激活和继发纤溶亢进 ,其在OSAHS患者心脑血管栓塞性并发症高发病率中起重要作用 ,并与夜间低氧血症密切相关 ;nCPAP治疗可有效逆转上述改变。     

The significance of platelet activation in ankylosing spondylitis

The objective of this study was to explore the significance of platelet activation in patients with ankylosing spondylitis (AS). Thirty-five AS patients and 15 normal controls were selected from November 2005 to October 2006. The number of CD62P- and CD63-positive cells were detected by flow cytometry. At the same time, the erythrocyte sedimentation rate (ESR), platelet count (PLT) and C-reactive protein (CRP) were determined in both groups. The percentage of CD62P-positive cell in AS patients (13.60 ± 7.64%) was significantly higher than that in control group (2.78 ± 1.04%; P < 0.01). The percentage of CD63-positive cell in AS patients (6.92 ± 4.16%) was significantly higher than that in control group (4.13 ± 1.85%; P < 0.05). The levels of CRP (20.18 ± 23.17 mg/l), PLT (259.54 ± 102.59 × 10⁹/l) and ESR (36.86 ± 31.23 mm/h) in AS patients were higher than those in normal controls, respectively (3.21 ± 2.18 mg/l, P < 0.01; 197.00 ± 55.70 × 10⁹/l, P < 0.01; 12.25 ± 5.05 mm/h, P < 0.05). Platelet activation may be a sign of AS exacerbation.     

Effects of FEIBA on platelet and leucocyte activation in severe haemophilia patients with inhibitors

Factor eight inhibitory bypassing agent (FEIBA) is used as a therapeutic option in haemophilia patients who have developed inhibitors. The measurement of thrombin generation has been applied to monitor the efficacy of FEIBA. However, a major concern about the clinical use of FEIBA is whether or not an increase in thrombin activity causes subsequent platelet activation and risk of thrombosis. Our aim is to evaluate whether FEIBA causes platelet and leucocyte activation in haemophilia patients with inhibitors. We evaluated the effects of FEIBA on platelet and leucocyte activity in correlation with thrombin generation. Initially, an in vitro study was conducted to evaluate the effects of FEIBA on platelet and leucocyte activity (using flow cytometry) using peripheral blood from normal volunteers. We then performed an ex vivo study looking at the effect of FEIBA on the above parameters in two haemophiliacs with high-titre inhibitors. A parallel study was also carried out ex vivo to evaluate thrombin generation using a thrombinoscope. FEIBA did not cause platelet or leucocyte activation in either the in vitro or ex vivo studies but showed a predictable increase in thrombin generation. Our study is the first one to address the effect of FEIBA on platelet and leucocyte function. We found no evidence of ‘systemic’ platelet activation. The findings suggest that whilst FEIBA improves global haemostasis, platelet activation is likely to be contained to the site of injury and systemic platelet activation, a previously feared consequence of FEIBA infusion that that may have contributed to thrombotic risk is absent.     

Markers of endothelial and in vivo platelet activation in patients with essential thrombocythemia and polycythemia vera

We investigated endothelial and in vivo platelet activation in a cohort of 52 patients with essential thrombocythemia (ET) and polycythemia vera (PV) before and after cytoreductive treatment, 22 healthy controls, and 17 patients with acute cerebrovascular ischemia (ACVI) and normal platelet counts. We measured platelet expression of CD62P and CD63 antigens and levels of soluble vascular cell adhesion molecule 1 (sVCAM-1). We found increased in vivo platelet activation in all patients with ET and PV, both before and after cytoreductive treatment, compared with controls. In patients with arterial thrombosis, platelet expression of CD62P, and in patients with erythromelalgia, expression of both markers was higher compared with expression in patients without thrombotic complications. In patients with ET and PV before and after treatment, sVCAM-1 expression was increased compared with expression in controls but also compared with expression in patients with ACVI and normal platelet counts. In patients with arterial thrombosis and erythromelalgia, in vivo platelet activation correlated with the level of sVCAM-1. Our findings indicated that in vivo platelet activation reflects intrinsic platelet defects in patients with ET and PV, persists after cytoreductive treatment, and results in endothelial damage, probably through release of angiogenic factors and/or activation of white blood cells.     

抗CD59对眼镜蛇毒因子介导的男性冠心病及健康成年人血小板的影响

目的 :研究补体激活对男性冠心病 (CHD)及健康成年人血小板的作用及抗CD5 9对该作用的影响。方法 :应用眼镜蛇毒因子 (CVF)活化血小板 ,通过测定男性CHD及健康成年人血小板的聚集及释放曲线 ,观察抗CD5 9应用前后CVF对血小板变形、聚集及释放功能的影响。结果 :CVF能诱导男性CHD及健康成年人血小板显著、持久的变形和释放 ,但不能诱导血小板聚集。当CVF浓度在 12 .5～ 5 0 .0g/L范围内 ,各组血小板的最大变形幅度与CVF浓度的对数呈线形相关 ,回归方程于CHD组 (36例 )为Y =2 8.717lgX - 19.798(r =0 .95 6 ,P<0 .0 1) ;健康成年人组为Y =2 6 .0 88lgX - 15 .5 81(r =0 .970 ,P <0 .0 1)。抗CD5 9能增强CVF诱导的血小板的最大变形幅度 ,并与剂量相关 ,其血小板最大变形幅度CHD组及健康成年人组分别为空白对照时的 1.39和1.36倍 (P <0 .0 1)。同时抗CD5 9也促进补体诱导的血小板ATP分泌增多。但两组间血小板对CVF及抗CD5 9的反应差异无统计学意义。结论 :活化的补体能诱导男性CHD及健康成年人血小板发生变形、释放 ,但不诱导血小板的聚集 ,且抗CD5 9能增强补体诱导的血小板功能改变     

替罗非班对急性冠脉综合征患者血小板活化度的影响

目的:评价GPⅡb/Ⅲa受体拮抗剂(盐酸替罗非班)对急性冠脉综合征(ACS)患者血小板活化度的影响及临床安全性。方法:受试组患者为来自2005年1月～2008年1月我院就诊的急性非ST段抬高型心肌梗塞和不稳定型心绞痛ACS患者,共126例,接受盐酸替罗非班持续泵入24～48h,根据年龄、性别选取同期就诊的稳定型心绞痛冠心病患者112例作为对照组。应用流式细胞仪分别对受试组和对照组患者CD61、CD62p、活化的GPⅡb/Ⅲa(PAC-1)的表达情况进行分析。结果:受试组CD61、PAC-1指标应用替罗非班1.5～2h,10h,48h比用药前明显降低(P均0.01),而对照组在治疗前后无明显变化;CD61仅用药1.5h时两组间比较有显著差异(P0.001);受试组PAC-1水平在治疗1.5～2h,10h时明显下降(P0.05),在48h又出现明显升高(P0.05)。结论:急性冠脉综合征患者应用GPⅡb/Ⅲa受体拮抗剂可改善长期的临床预后,同时我们的研究也显示替罗非班治疗急性冠脉综合症是有效的和安全的。     

Relationship and significance between anti-β2-glycoprotein I antibodies and platelet activation state in patients with ulcerative colitis

AIM:To study the relationship between anti-β2-glycoproteinⅠ(aβ2GPⅠ) antibodies and platelet activation state in patients with ulcerative colitis (UC)and its significance.METHODS:Peripheral blood samples were collected from 56 UC patients (34 males and 22 females,aged 43.5 years,range 21-66 years),including 36 at active stage and 20 at remission stage,and 25 sex-and age-matched controls.The level of aβ2GPⅠwas measured by ELISA.The platelet activation markers,platelet activation complex-I (PAC- I) and P-selectin (CD62P) were detected by flow cytometry.RESULTS:The A value for IgG aβzGPⅠin the active UC group was 0.61±0.13,significantly higher than that in the remittent UC and control groups (0.50±0.13 and 0.22±0.14,P＜0.01).There was a significant difference between the two groups (P＜0.01).The A value for IgM aβ2GPⅠin the active and remittent UC groups was 0.43±0.13 and 0.38±0.12,significantly higher than that in the control group (0.20±0.12,P＜0.01).However,there was no significant difference between the two groups (P＞0.05).The PAC-I positive rate for the active and remittent UC groups was 30.6%±7.6% and 19.6%±7.8% respectively,significantly higher than that for the control group (6.3%±1.7%,P＜0.01).There was a significant difference between the two groups (P＜0.01).The CD62P positive rate for the active and remittent UC groups was 45.0%±8.8% and 31.9%±7.8% respectively,significantly higher than that for the control group (9.2%±2.7%,P＜0.01).There was a significant difference between the two groups (P＜0.01).In the active UC group,the more severe the state of illness was,the higher the A value for IgG aβ2GPⅠwas,and the positive rate for PAC-I and CD62P was positively correlated with the state of illness (Faβ2GPⅠ= 3.679,P＜0.05;FPAC-I (%) = 5.346,P＜0.01;and FCD62P (%) = 5.418,P＜0.01).Meanwhile,in the same state of illness,the A value for IgG aβ2GPⅠwas positively correlated to the positive rates for PAC-I and CD62P.CONCLUSION:aβ2GPⅠlevel,platelet activation state and their relationship of them are closely correlated with the pathogenesis and development of UC.     

The significance of platelet activation in rheumatoid arthritis

We evaluated the significance of platelet activation in patients with rheumatoid arthritis (RA). The expression of CD62P and CD63 by platelets was determined using flow cytometry in 18 active RA patients, 10 remission RA and 15 normal controls. Meanwhile, the erythrocyte sedimentation rate (ESR) and C-reactive protein was also determined in all groups. The expression of CD62P in active RA patients (11.88 ± 2.47%) was significantly higher than that in remission RA group (2.85 ± 1.60%; P < 0.01) and control group (2.78 ± 1.04%; P < 0.01). The expression of CD63 in active RA patients (9.90 ± 3.02%) was significantly higher than that in remission RA group (4.11 ± 2.00%; P < 0.01) and control group (4.13 ± 1.85%; P < 0.01). The level of CRP (54.33 ± 23.35 mg/l) and ESR (86.06 ± 33.67 mm/h) in active RA patients was higher than that in remission RA group (2.55 ± 1.01 mg/l, 14.70 ± 4.57 mm/h; P < 0.01 for both) and normal control group (3.21 ± 2.18 mg/l, 12.25 ± 5.05 mm/h; P < 0.01 for both). There was a positive correlation between CD62P and ESR (r = 0.5224, P < 0.01) and also a positive correlation between CD62P and CRP (r = 0.7048, P < 0.01) as well as between CD63 and ESR (r = 0.4476, P < 0.05) but no correlation between CD63 and CRP. Platelet activation may be a sign of RA exacerbation.     

急性缺血性脑卒中患者血小板活化水平与全身炎症反应综合征的相关性

目的 动态观察缺血性脑卒中患者外周血血小板活化水平,分析血小板活化与伴发全身炎症反应综合征(SIRS)的关系. 方法 动态监测缺血性脑卒中患者外周血血小板平均内含物浓度(MPC)和白细胞计数及发病早期血浆纤维蛋白原(FIB)和血小板聚集率等指标,比较SIRS组与无SIRS组患者的各项参数,对发病早期MPC水平与SIRS评分以及当日白细胞计数、FIB和血小板聚集率等进行相关性分析. 结果 MPC水平在发病第1天略有下降,从第2天至病程第45天一直保持较低水平,均值在229～242 g/L之间波动.SIRS组与无SIRS组比较发病早期FIB差异无统计学意义(t=1.835,P=0.07),其余各指标差异有统计学意义(均为P＜0.01).发病前3 d MPC水平与SIRS评分呈显著负相关(相关系数分别为-0.392、-0.376、-0.341,t值分别为3.484、3.405、3.125,均为P＜0.01),而与当日白细胞计数、FIB及血小板聚集率无相关性(P＞0.05). 结论 缺血性脑卒中患者整个病程期间血小板一直保持较活化的状态,伴发SIRS的患者发病早期血小板活化水平、白细胞计数和血小板聚集率均高于未伴发SIRS的患者.早期的血小板活化水平提示SIRS的严重程度,血小板活化可能是SIRS的独立危险因素.     

培哚普利、非洛地平缓释片对老年原发性高血压患者血小板活化及纤溶活性的影响

目的:探讨培哚普利及非洛地平缓释片对老年原发性高血压(EH)患者血小板活化及纤溶活性的影响.方法:采用放射免疫法及发色底物法测定55例老年Ⅰ、Ⅱ期EH患者分别应用培哚普利(Ⅰ组28例)及非洛地平缓释片(Ⅱ组27例)治疗前后活化血小板α颗粒膜蛋白(GMP-140)、组织型纤溶酶原激活剂(t-PA)及其抑制物(PAI-1)、血管紧张素Ⅱ(AngⅡ)及醛固酮(Ald)水平.结果:治疗后,两组患者GMP-140含量及PAI-1活性均显著下降(P分别＜0.01,0.05),t-PA活性显著升高(P＜0.05);Ⅰ组患者AngⅡ及Ald显著下降(P＜0.01),Ⅱ组患者AngⅡ及Ald未见明显变化(P＞0.05).治疗前,55例EH患者GMP-140含量与PAI-1水平呈显著正相关(r=0.407 6,P＜0.01),二者与AngⅡ水平均呈显著正相关(r=0.416 2,0.4237,P＜0.01).结论:以上结果提示,培哚普利降低体内AngⅡ水平是其抑制老年EH患者血小板活化、改善纤溶活性的机制之一,而非洛地平缓释片抑制血小板活化、改善纤溶活性的作用与肾素-血管紧张素系统无关.     

Change of platelet activation markers using flow cytometry in patients with hematology/oncology disorders after transfusion

In spite of the frequent need of platelet transfusions, there is limited information on the association of platelet activation markers, in transfused patients with hematology/oncology disorders, with platelet function using flow cytometry. The goal of this study was to evaluate the changes of PAC-1 binding and CD62P expression, with or without agonists in patients after transfusions. Twenty-eight whole blood samples were obtained from 24 patients admitted to the department of Hematology & Oncology and transfused with platelets; these samples were compared to 30 healthy controls. Whole blood samples, either with or without agonists, such as 20 µM adenosine diphosphate (ADP) or 100 µM thrombin receptor activating peptide (TRAP), were stained with the fluorescein conjugated monoclonal antibodies PAC-1 or CD62P. Then, the percent expression for each marker was analysed using flow cytometry. ADP and TRAP induced an increased percentage of CD62P expression and PAC-1 binding after platelet transfusions compared to the samples studied before transfusion, and these findings were lower than those of the healthy controls. However, the expression of platelets without the agonists was not significantly changed, despite the transfusions. Therefore, agonist-induced platelet activation markers, studied by flow cytometry, appear to be more useful for the evaluation of platelet function after transfusions than platelet activation markers without agonists.     

Effects of low osmolar contrast (iomeprol) on haemorheology and platelet activation in patients with coronary artery disease

Background Non-ionic low osmolar contrast agents are widely used during coronary angiography. As these agents cause activation of thrombotic pathways in vitro, this may have potentially significant clinical impact. However, limited evidence exists as to their in vivo effects from selective coronary cannulation. Methods We initially performed an in vitro experiment to assess the effect of serial contrast (Iomeprol 300, Bracco) dilution on platelet indices [mean platelet count (MPC), platelet volume (MPV), platelet granularity (MPG)]. The in vivo effect of contrast injection on platelet activation markers [soluble P-selectin (sPsel), soluble CD40 ligand (sCD40L)], MPC, MPV, MPG, haemoglobin and haematocrit was subsequently determined in 35 patients (mean age 58 ± 11; 22 males) undergoing cardiac catheterisaton. Results No significant in vitro effect of contrast on MPC or MPV was seen but there was a significant increase in MPG (p = 0.40, 0.10 and 0.01, respectively). In the in vivo study, there was a reduction in mean haemoglobin and haematocrit levels, suggesting an average increase in plasma volume of 6.5 ± 5.8%. The in vivo effect of Iomeprol was associated with an unadjusted reduction in sPsel concentrations (p = 0.04) and MPV (p < 0.05), with denser platelets (p < 0.05). There was no difference in MPC or sCD40L concentration (both p = NS). After adjustment for the haemodilution effect, no significant reduction in P-sel levels was seen with contrast (p = 0.27), although the adjusted post-contrast change in MPG (p = 0.01), MPC (p = 0.01) and sCD40L (p < 0.05) levels were significant. Conclusion Low osmolar contrast led to a minimal effect on soluble and physical indices of platelets within the coronary artery, primarily due to plasma volume expansion.     

左心房血栓形成中血小板活化作用的研究

目的:检测风湿性二尖瓣狭窄合并心房颤动的患者血浆P-选择素浓度变化,研究P-选择素蛋白及其在左心耳组织中的基因表达情况,探讨血小板活化在左心房血栓形成中的作用。方法:风湿性二尖瓣狭窄合并心房颤动患者80例(血栓组40例,无血栓组40例),健康人对照组15例(心脏移植供体10例取左心耳)。采用酶联免疫吸附剂测定法检测左心房及外周血浆P-选择素浓度。左心耳组织切片免疫组化染色,光镜下观察。实时荧光定量PCR对左心耳组织P-选择素基因表达进行定量研究。结果:与无血栓组相比,血栓组心房颤动时间更长、二尖瓣口面积更小、左心房内径更大,两组左心房内径均大于对照组。左心房与外周血P-选择素浓度差异均无显著性,血栓组外周血P-选择素浓度高于无血栓组,两组均高于对照组,免疫组化染色显示血管性血友病因子(vWF)和p-选择素蛋白在心内膜和心肌细胞中均有表达。左心耳组织P-选择素基因相对于正常左心耳组织的表达量,各组间差异无统计学意义性。结论:风湿性二尖瓣狭窄合并心房颤动患者血浆P-选择素浓度水平升高,外周血P-选择素浓度可代表左心房血中的浓度水平,血浆P-选择素浓度升高与P-选择素基因表达水平改变无关。     

大动脉粥样硬化性卒中与脑小血管病患者血小板PAC-1、CD62P阳性率和血清血小板因子4水平的差异

目的 探讨脑小血管病及其亚型与大动脉粥样硬化性卒中患者血小板膜糖蛋白PAC-1和CD62P表达的差异,并对传统血小板活化标记物和炎症趋化因子血小板因子4(platelet factor 4,PF4)进行比较.方法 分别采用流式细胞术和酶联免疫吸附法测定30例大动脉粥样硬化性卒中患者、45例脑小血管病患者和30例对照者外...     

In Vivo and protease-activated receptor-1-mediated platelet activation in patients presenting for cardiac catheterization

Pathways of platelet activation that are not targeted by current antithrombotic therapy may be crucial for the development of ischemic events in patients undergoing coronary angiography. We therefore investigated whether in vivo and thrombin receptor activating peptide (TRAP)-stimulated platelet activation and monocyte-platelet aggregate (MPA) levels can serve as independent risk markers for adverse outcomes in aspirin-treated patients presenting for cardiac catheterization. In vivo and TRAP-stimulated platelet surface P-selectin, activated glycoprotein IIb/IIIa (GPIIb/IIIa) and MPA levels were determined in 682 consecutive patients undergoing cardiac catheterization and in 47 healthy controls. Two-year follow-up data were obtained from 562 patients. In vivo platelet surface P-selectin, activated GPIIb/IIIa and MPA levels were significantly higher in patients with angiographically-proven coronary artery disease than in healthy controls (all p≤0.02). Patients with an acute coronary syndrome (ACS; n=125) had significantly higher levels of in vivo MPA than patients without ACS (n=437; p=0.01). In the overall study population (n=562) the surface expression of P-selectin and activated GPIIb/IIIa, and the levels of MPA in vivo and in response to TRAP were similar in patients without and with subsequent ischemic events (all p>0.05). Similar results were obtained when only patients with angiographically-proven coronary artery disease (n=459), stent implantation (n=205) or ACS (n=125) were analyzed. Receiver-operating characteristic curve analyses did not reveal cut-off values for P-selectin, activated GPIIb/IIIa, and MPA levels for the prediction of ischemic events. In conclusion, in vivo and TRAP-stimulated platelet activation and MPA levels did not predict adverse ischemic outcomes in aspirin-treated patients presenting for cardiac catheterization.     

血小板活化与心血管疾病

血小板活化在心血管疾病的发生发展中起着非常重要的作用。本文对血小板活化的分子标志物与心血管疾病的关系进行综述。     

静脉注射肝素钠和达肝素钠对血小板激活影响的对比研究

目的了解肝素钠(UFH)与低分子肝素(LWMH)对血小板激活的影响有无差别。方法59例新入院冠心病患者,随机分为UFH组和达肝素钠组,分别于基线状态下、给药(UFH或达肝素钠5000 IU)后30 min、1 h取血,测定CD62P、GPⅡb/Ⅲa、血管性假血友病因子(vWF)。结果UFH组静脉给药后30 min GPⅡb/Ⅲa、血浆CD62P及vWF的水平均明显高于给药前(P<0.05);给药后1 h GPⅡb/Ⅲa及vWF的水平仍显著高于给药前(P<0.05);血浆CD62P的水平虽仍较给药前增高,但差异无统计学意义(P>0.05)。静脉给予达肝素钠5000 IU30min后GPⅡb/Ⅲa及血浆CD62P的水平均明显高于给药前(P<0.05),但在给药后1 h即与基线值差异无统计学意义(P>0.05);vWF的水平在给予达肝素钠后30 min、1 h均与给药前相比,差异无统计学意义(P>0.05)。结论静脉注射UFH及达肝素钠均可激活血小板;但达肝素钠对血小板活化的影响较小。