Bruder J L, Hsieh T, Lerea K M, Olson S C, Wu J M
Department of Biochemistry, New York Medical College, Valhalla, New York 10595, USA.
BMC Cell Biol. 2001;2:1. doi: 10.1186/1471-2121-2-1. Epub 2001 Jan 29.
Atherosclerosis and coronary heart disease (CHD) are significant contributors to morbidity and mortality in developed countries. A noted exception is the low mortality of CHD in France, particularly the southwest region. This phenomenon, commonly referred to as the French paradox, may be associated with high consumption of red wine. We investigate whether the cardioprotective activity of red wine may involve the grape skin-derived polyphenol, resveratrol. We further test the possibility that resveratrol acts by modulating structural and functional changes in endothelial cells lining the blood vessel wall.
Bovine pulmonary artery endothelial cells (BPAEC) were incubated with resveratrol, with and without concurrent exposure to simulated arterial shear stress. Resveratrol significantly affected proliferation and shape of BPAEC; growth was suppressed and cells became elongated, based on morphologic analysis of rhodamine-conjugated phalloidin stained F-actin by confocal microscopy. Using selective signaling inhibitors, we showed that the resveratrol-induced cellular phenotype was dependent on intracellular calcium and tyrosine kinase activities, and assembly of actin microfilaments and microtubules, but was unrelated to PKC activity. Exposure to simulated arterial flow revealed that, whereas controls cells easily detached from the culture support in a time-dependent manner, resulting in total cell loss after a 5 min challenge with simulated arterial flow conditions, a significant percentage of the treated cells remained attached to the cultured plastic coverslips under identical experimental conditions, suggesting that they adhered more strongly to the surface. Western blot analysis shows that whereas cells treated with 25 microM and 100 microM resveratrol had no change in total ERK1/2, treatment did result in an increase in phosphorylated ERK1/2, which probably involved stabilization of the active enzyme. An increase in nitric oxide synthase expression was detected as early as 6 h and persisted for up to 4 days of treatment.
Results of our studies show that resveratrol interacts with endothelial cells in vitro to elicit morphological and structural changes; the observed changes support the interpretation that resveratrol acts as a cardioprotective agent.
动脉粥样硬化和冠心病（CHD）是发达国家发病和死亡的主要原因。一个显著的例外是法国，尤其是西南部地区冠心病的低死亡率。这种现象通常被称为法国悖论，可能与红酒的高消费量有关。我们研究红酒的心脏保护活性是否可能涉及葡萄皮来源的多酚白藜芦醇。我们进一步测试白藜芦醇是否通过调节血管壁内衬内皮细胞的结构和功能变化来发挥作用。
牛肺动脉内皮细胞（BPAEC）与白藜芦醇一起孵育，同时或不同时暴露于模拟动脉剪切应力。白藜芦醇显著影响BPAEC的增殖和形态；基于共聚焦显微镜对罗丹明标记的鬼笔环肽染色的F-肌动蛋白的形态学分析，生长受到抑制，细胞变得细长。使用选择性信号抑制剂，我们表明白藜芦醇诱导的细胞表型依赖于细胞内钙和酪氨酸激酶活性，以及肌动蛋白微丝和微管的组装，但与蛋白激酶C活性无关。暴露于模拟动脉血流显示，对照细胞在时间依赖性方式下容易从培养支持物上脱落，在模拟动脉血流条件下5分钟的刺激后导致细胞完全丧失，而在相同实验条件下，相当比例的处理细胞仍附着在培养的塑料盖玻片上，表明它们与表面的粘附更强。蛋白质印迹分析表明，用25 microM和100 microM白藜芦醇处理的细胞总ERK1/2没有变化，但处理确实导致磷酸化ERK1/2增加，这可能涉及活性酶的稳定。早在处理6小时就检测到一氧化氮合酶表达增加，并持续长达4天。
我们的研究结果表明，白藜芦醇在体外与内皮细胞相互作用，引发形态和结构变化；观察到的变化支持白藜芦醇作为心脏保护剂的解释。
