抗磷脂综合征患者的血清可诱导内皮细胞中的黏附分子。

Serum of patients with antiphospholipid syndrome induces adhesion molecules in endothelial cells.

作者信息

Engel Bettina, Müller Gregor, Roch Beate, Schröder Hans-Egbert, Aringer Martin, Bornstein Stefan R, Morawietz Henning

机构信息

Division of Vascular Endothelium and Microcirculation, Department of Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany; Institut für Hausarztmedizin, Medizinische Fakultät Bonn, Bonn, Germany.

Division of Vascular Endothelium and Microcirculation, Department of Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.

出版信息

Atheroscler Suppl. 2017 Nov;30:141-148. doi: 10.1016/j.atherosclerosissup.2017.05.039. Epub 2017 Jun 2.

DOI:10.1016/j.atherosclerosissup.2017.05.039

PMID:29096830

Abstract

OBJECTIVE

The antiphospholipid syndrome (APS) is a systemic auto-immune disease with an unclear pathophysiology. The aim of our study was to understand the development of APS on a cellular level. Therefore, we analyzed the influence of human serum of APS patients on endothelial expression of specific genes and proteins in comparison to a control group.

METHODS

In this study, we analyzed the expression of ICAM-1, VCAM-1, E-selectin and annexin V in primary cultures of human umbilical vein endothelial cells (HUVEC) in response to 10% (v/v) serum of control patients (n = 6), patients with systemic lupus erythematosus (SLE) and no APS (n = 4) or APS patients (n = 9) for 24 h. Total RNA was prepared from confluent endothelial cell layers and mRNA expression of ICAM-1, VCAM-1 and E-selectin was analyzed by reverse transcription polymerase-chain reaction (RT-PCR). The protein expression was determined by Western blot. Serum protein concentrations of soluble forms of adhesion molecules sICAM-1 and sVCAM-1 were quantified by ELISA. Gene expression data were correlated with clinical parameters.

RESULTS

The mRNA expression of ICAM-1 was increased in cells incubated with serum from APS patients (166 ± 22% of control; P = 0.023). Serum of patients with (SLE)/no APS caused a 1.4-fold higher ICAM-1 mRNA level. Western blot analysis showed an increase in protein expression of adhesion molecules ICAM-1 (260 ± 49%; P = 0.011) and VCAM-1 (357 ± 97%; P = 0.023) in cells that were incubated with serum from APS patients. Plasma analysis showed elevated levels of sVCAM-1 in APS patients (189 ± 34%; P = 0.045) compared to the levels measured in the control group. The sVCAM-1 plasma level was correlating with the frequency of abortions.

CONCLUSION

An augmented expression of endothelial adhesion molecules is involved in the pathophysiology of patients with antiphospholipid syndrome.

摘要

目的

抗磷脂综合征（APS）是一种病理生理学尚不明确的系统性自身免疫性疾病。我们研究的目的是在细胞水平上了解APS的发病机制。因此，我们分析了APS患者血清与对照组相比对特定基因和蛋白质在内皮细胞中的表达的影响。

方法

在本研究中，我们分析了人脐静脉内皮细胞（HUVEC）原代培养物中细胞间黏附分子-1（ICAM-1）、血管细胞黏附分子-1（VCAM-1）、E-选择素和膜联蛋白V的表达，这些细胞分别用10%（v/v）的对照患者血清（n = 6）、无APS的系统性红斑狼疮（SLE）患者血清（n = 4）或APS患者血清（n = 9）处理24小时。从汇合的内皮细胞层中提取总RNA，并通过逆转录聚合酶链反应（RT-PCR）分析ICAM-1、VCAM-1和E-选择素的mRNA表达。通过蛋白质印迹法测定蛋白质表达。通过酶联免疫吸附测定（ELISA）对可溶性黏附分子sICAM-1和sVCAM-1的血清蛋白浓度进行定量。基因表达数据与临床参数相关。

结果

与用APS患者血清孵育的细胞相比，ICAM-1的mRNA表达增加（为对照的166±22%；P = 0.023）。SLE/无APS患者的血清使ICAM-1 mRNA水平升高1.4倍。蛋白质印迹分析显示，用APS患者血清孵育的细胞中，黏附分子ICAM-1（260±49%；P = 0.011）和VCAM-1（357±97%；P = 0.023）的蛋白质表达增加。血浆分析显示，与对照组相比，APS患者的sVCAM-1水平升高（189±34%；P = 0.045）。sVCAM-1血浆水平与流产频率相关。