Medizinische Klinik und Poliklinik I, Campus Grosshadern, Ludwig-Maximilians-Universität, München, Germany.
Microvasc Res. 2009 Dec;78(3):364-9. doi: 10.1016/j.mvr.2009.07.008. Epub 2009 Aug 5.
An increasing number of studies have examined the role of emotional stress and coronary heart disease; the underlying pathophysiology is still poorly understood. The present study was designed to evaluate the relationship between acute (epi- and norepinephrine) and chronic stress hormones (dexamethasone, beta-endorphin, corticotropin releasing hormone) and endothelial dysfunction.
Human microvascular endothelial cells were incubated with stress hormones for 6 and 24 h. ET-1 release and ADMA were quantified via ELISA, NO release by using cell permeable 4.5-diaminofluorescein diacetate (DAF2-DA), oxidative stress fluometrically by the ROS-sensitive carboxy-H2-DCFDA method, mitochondrial metabolic activity by using the colorimetric assay WST-1, ET-1 receptor type A (ET(A)R) protein expression by Western blot, and cell proliferation activity was assessed by the colorimetric assay BrdU.
With respect to analysed acute and chronic stress hormones, ET-1 release was significantly increased. Likewise, protein expression was enhanced after long term incubation (24 h) with norepinephrine and dexamethasone. In contrast, endothelial NO-levels were only influenced by short term stimulation of dexamethasone (upregulation of NO release) and norepinephrine (downregulation of NO release), whereas modified NO concentration mimics altered mitochondrial metabolic activity. Unexpectedly, both oxidative stress and cell proliferation were not modified by stress hormones.
Results suggest that acute and chronic stress hormones induce a significant ET-1 release whereas NO release remained mainly unchanged. The imbalance of pro- and antiatherosclerotic factors may play a pivotal role in the initiation of stress-related endothelial dysfunction up to myocardial infarction.
越来越多的研究探讨了情绪应激与冠心病之间的关系，但其潜在的病理生理学机制仍知之甚少。本研究旨在评估急性（肾上腺素和去甲肾上腺素）和慢性应激激素（地塞米松、β-内啡肽、促肾上腺皮质激素释放激素）与内皮功能障碍之间的关系。
将人微血管内皮细胞与应激激素孵育 6 和 24 小时。通过 ELISA 定量测定 ET-1 释放和 ADMA，通过细胞通透性 4.5-二氨基荧光素二乙酸酯（DAF2-DA）测定 NO 释放，通过 ROS 敏感羧基-H2-DCFDA 方法荧光法测定氧化应激，通过比色法 WST-1 测定线粒体代谢活性，通过 Western blot 测定 ET-1 受体 A（ET(A)R）蛋白表达，通过 BrdU 比色法评估细胞增殖活性。
就分析的急性和慢性应激激素而言，ET-1 的释放显著增加。同样，在长时间孵育（24 小时）后，去甲肾上腺素和地塞米松的蛋白表达也增强。相比之下，只有地塞米松（上调 NO 释放）和去甲肾上腺素（下调 NO 释放）的短期刺激才会影响内皮细胞的 NO 水平，而修饰后的 NO 浓度则模拟了改变的线粒体代谢活性。出乎意料的是，应激激素并未改变氧化应激和细胞增殖。
研究结果表明，急性和慢性应激激素可引起显著的 ET-1 释放，而 NO 释放基本保持不变。促动脉粥样硬化和抗动脉粥样硬化因子的失衡可能在应激相关内皮功能障碍的发生直至心肌梗死中发挥关键作用。
