Protein C activation in endothelial cells is inhibited under the presence of anticardiolipin antibody in ischemic stroke patients

缺血性中风患者抗心磷脂抗体存在下，内皮细胞中的蛋白 C 活化受到抑制

• 批准号: 07670731
• 负责人: KITAGAWA Yasuhisa
• 金额: $ 0.45万
• 依托单位: Tokai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670731/
• 关键词: anticardiolipin antibody; cerebral infaretion; protein C; endothelial cell; antiphospholipid antibody; thrombosis; プロテインC

INTRODUCTION : Anticardiolipin antibody (aCL) is considered to be one of the contributory factors in the development of cerebral infarction. A number of sites exist in the coagulation system where interaction with aCL may lead to thrombosis. However, the precise mechanism of action of aCL remains unknown. Human activated protein C functions as a potent anticoagulant in human plasma by inhibiting the activity of coagulation cofactors Va and VIIIa. The purpose of the present study was to investigate the effect of serum IgG fraction obtained from ischemic stroke patients on the rate of protein C activation in cultured human endothelial cells from the umbilical cord veins.SUBJECTS AND METHODS : The rate of protein C activation was examined in the following groups. We had 10 ischemic stroke patients with positive IgG aCL.These comprised 7 patients with cerebral infarction related SLE (Group 1) and 3 patients without an immunological background including SLE (Group 2). We also examined in 6 … More non-SLE related ischemic stroke patients without aCL (Group 3), 14 SLE patients with aCL who did not develop ischemic stroke (Group 4), 6 SLE patients without stroke and aCL (Group 5) and 6 normal controls (Group 6). Confluent endothelial cells were incubated with buffer or IgG obtained from the patients (7 mg/ml) for 1 hour at 37ﾟC.Purified human protein C (1 nM/well) and thrombin (0.04 nM/well) were added. After 2 hours incubation at 37ﾟC,the rate of protein C activation was determined by amidolytic assay employing P-Glu-Pro-Arg-MNA as a substrate. The degree of activation of protein C was expressed as the percent change in the measured absorbance between the presence and absence of the IgG fraction. A negative percent change indicated inhibition of protein C activation.RESULTS : In SLE related ischemic stroke with positive aCL (Group 1), 5 of the 7 patients revealed a negative percent change which indicated inhibition of the rate of protein C activation by addition of the IgG fraction. The mean percent change of absorbance in these 7 patients was -7.2% (range, from -21.4% to+4.5%). In the 3 non-SLE stroke patients with positive aCL (Group 2), the percent changes in absorbance were -2.0%, -9.8% and -6.6%, respectively. On the other hand, IgG fraction did not appreciably influence the activation of protein C in the patients of Group 3 (the mean parcent change of absorbance, +0.1%), Group 4 (-0.5%) and Group 5 (+0.3%). In the normal controls, the mean percent change in the measured absorbance was +0.6%.CONCLUSION : Our results appear to indicate that aCL in IgG fractions can interfere with interactions among thrombin, protein C and thrombomodulin on the surface of endothelial cells. Impairment of the rate of activation of protein C could play a role in the development of ischemic stroke. Less

前言：抗心磷脂抗体（aCL）被认为是脑梗死发生的促发因素之一。凝血系统中存在许多与aCL相互作用可能导致血栓形成的部位。然而，ACL的确切作用机制仍不清楚。人活化蛋白C通过抑制凝血辅因子Va和VIIIa的活性，在人血浆中起到强效抗凝剂的作用。本研究的目的是探讨缺血性中风患者血清IgG组分对蛋白C活化率在培养的人脐静脉内皮细胞从脐带vein.SUBJECTS和方法：蛋白C活化率的影响进行了检查，在以下各组。10例缺血性脑卒中患者IgG aCL阳性，其中7例为脑梗死相关SLE患者（第1组），3例无免疫学背景（包括SLE）患者（第2组）。我们还在6 ...更多信息 非SLE相关的缺血性脑卒中患者无aCL（第3组），14例SLE伴aCL但未发生缺血性脑卒中（第4组），6例SLE患者无脑卒中和aCL（第5组），6例正常对照（第6组）。将汇合的内皮细胞与从患者获得的缓冲液或IgG（7 mg/ml）在37 ℃下孵育1小时。加入纯化的人蛋白C（1 nM/孔）和凝血酶（0.04nM/孔）。在37 ℃孵育2小时后，采用P-Glu-Pro-Arg-MNA作为底物，通过酰胺分解试验测定蛋白C活化速率。蛋白C的活化程度表示为存在和不存在IgG级分之间测量的吸光度的百分比变化。A负百分比变化表示蛋白C activation.Results的抑制：在SLE相关的缺血性中风与阳性ACL（组1），5 7例患者显示了一个负的百分比变化，这表明抑制蛋白C激活的速度，另外的IgG部分。这7例患者的吸光度平均百分比变化为-7.2%（范围为-21.4%至+4.5%）。在3例aCL阳性的非SLE卒中患者（第2组）中，吸光度的百分比变化分别为-2.0%、-9.8%和-6.6%。另一方面，IgG组分对第3组（吸光度的平均百分比变化，+0.1%）、第4组（-0.5%）和第5组（+0.3%）患者中蛋白C的活化没有明显影响。在正常对照组中，测得的吸光度的平均百分比变化为+0.6%。结论：IgG组分中的aCL可干扰内皮细胞表面凝血酶、蛋白C和血栓调节蛋白之间的相互作用。蛋白C活化率的降低可能在缺血性卒中的发生中起作用。少

项目成果

Kitagawa Y et al: "Protein C activation in endothelial cell is inhibited under the presence of anticadiolipin antibody in ischemic Stroke patients" J Cereral Blood Flow Metabol. 15 (suppl 1). S684- (1995)

Kitakawa Y 等人：“缺血性中风患者中，内皮细胞中的蛋白 C 活化在抗心磷脂抗体的存在下受到抑制”J Cereral Blood Flow Metabol。