Voisard R, Fischer R, Osswald M, Voglic S, Baur R, Susa M, Koenig W, Hombach V
Department of Internal Medicine II, Division of Cardiology, University of Ulm, Germany.
Circulation. 2001 Mar 27;103(12):1688-94. doi: 10.1161/01.cir.103.12.1688.
Leukocyte attack (LA) and the triggered reactive proliferation of smooth muscle cells (SMCs) are key events for the development of early atherosclerosis and restenosis. In the present study, we used a 3D human coronary in vitro model of LA (3DLA model) to examine the effect of high-dose aspirin on the adhesion and chemotaxis of leukocytes and the reactive proliferative response of SMCs.
For dose-finding, the effect of aspirin (1, 2, 5, and 10 mmol/L) on the tumor necrosis factor-alpha-induced upregulation of intercellular adhesion molecule-1 was analyzed in monocultures of human coronary endothelial cells (HCAEC) and the SMCs of the human coronary media (HCMSMC). In cytoflow and Northern blot experiments, the expression of intercellular adhesion molecule-1 was slightly reduced after incubation with 5 mmol/L aspirin, and strong inhibition was found after incubation with 10 mmol/L. In 3DLA models, HCAECs and HCMSMCs were cultured on both sides of a porous filter. For LA, human monocytes or CD4(+) lymphocytes were seeded on the HCAEC side of the 3DLA unit. A dose of 5 mmol/L aspirin inhibited the adherence of monocytes or CD4(+) lymphocytes by 50% (P:<0.01) and the chemotaxis of monocytes by 90% (P:<0.01). The reactive proliferative response of cocultured HCMSMCs after LA, as measured by the uptake of bromodeoxyuridine, was significantly reduced by 83% after selective monocyte attack (P:<0.001) and by 42% after selective CD4(+) lymphocyte attack (P:<0.05).
A local concentration of 5 mmol/L aspirin should be accepted as the lowest rational concentration for the beneficial in vitro effects of high-dose aspirin to be reproduced in clinical studies.
白细胞攻击（LA）和平滑肌细胞（SMC）的触发反应性增殖是早期动脉粥样硬化和再狭窄发展的关键事件。在本研究中，我们使用三维人冠状动脉体外LA模型（3DLA模型）来研究高剂量阿司匹林对白细胞黏附和趋化性以及SMC反应性增殖反应的影响。
为了确定剂量，在人冠状动脉内皮细胞（HCAEC）和人冠状动脉中膜平滑肌细胞（HCMSMC）的单培养物中分析了阿司匹林（1、2、5和10 mmol/L）对肿瘤坏死因子-α诱导的细胞间黏附分子-1上调的影响。在细胞流式和Northern印迹实验中，与5 mmol/L阿司匹林孵育后，细胞间黏附分子-1的表达略有降低，与10 mmol/L孵育后发现有强烈抑制作用。在3DLA模型中，HCAEC和HCMSMC在多孔滤膜的两侧培养。对于LA，将人单核细胞或CD4(+)淋巴细胞接种在3DLA单元的HCAEC一侧。5 mmol/L的阿司匹林剂量可使单核细胞或CD4(+)淋巴细胞的黏附减少50%（P<0.01），单核细胞的趋化性减少90%（P<0.01）。通过溴脱氧尿苷摄取测量，LA后共培养的HCMSMC的反应性增殖反应在选择性单核细胞攻击后显著降低83%（P<0.001），在选择性CD4(+)淋巴细胞攻击后降低42%（P<0.05）。
5 mmol/L的局部阿司匹林浓度应被视为在临床研究中重现高剂量阿司匹林体外有益作用的最低合理浓度。
