Schraml Elisabeth, Quan Ping, Stelzer Ingeborg, Fuchs Robert, Skalicky Monika, Viidik Andrus, Schauenstein Konrad
Institute of Pathophysiology, Center of Molecular Medicine, Medical University of Graz, Heinrichstrasse 31A, 8010 Graz, Austria.
Exp Gerontol. 2007 Nov;42(11):1072-8. doi: 10.1016/j.exger.2007.08.003. Epub 2007 Aug 11.
Reactive oxygen species (ROS) play important roles in cellular senescence and organismic aging. Furthermore, they have been implicated in some of the adverse effects of chronic stress due to elevated peripheral levels of catecholamines. Here, we applied three different techniques to individually compare the systemic and intracellular oxidative stress in aged (23 months) and young (5 months) Sprague-Dawley rats, and in young rats treated for 12 or 24 h with norepinephrine (NE). Thiol groups of blood serum proteins (RSH) were determined by means of Ellman's reaction. Intracellular ROS were assessed in spleen cells and peripheral blood lymphocytes (PBL) by carbonylation of cellular (spleen) proteins as determined by immunoblotting (Oxyblot) and/or by means of 2',7'-dichlorofluorescein (DCF) fluorescence. As compared to the young, untreated controls, both old rats and NE treated young rats showed similarly lowered RSH values paralleled by elevated intracellular ROS levels or enhanced Oxyblot signals. Individual RSH values were highly significantly, negatively correlated with respective Oxyblot data as well as with DCF fluorescence. The results confirm the roles of ROS in aging and adrenergic stress in the rat model, and suggest that the decrease in RSH of blood serum may be taken as a valid indicator for the enhanced oxidative stress in lymphocytes.
活性氧（ROS）在细胞衰老和机体老化过程中发挥着重要作用。此外，由于外周儿茶酚胺水平升高，它们还与慢性应激的一些不良影响有关。在此，我们应用三种不同技术，分别比较老年（23个月）和年轻（5个月）的Sprague-Dawley大鼠，以及用去甲肾上腺素（NE）处理12或24小时的年轻大鼠的全身和细胞内氧化应激。通过埃尔曼反应测定血清蛋白的巯基（RSH）。通过免疫印迹法（Oxyblot）测定细胞（脾脏）蛋白的羰基化和/或通过2'，7'-二氯荧光素（DCF）荧光，评估脾细胞和外周血淋巴细胞（PBL）中的细胞内ROS。与未处理的年轻对照相比，老年大鼠和经NE处理的年轻大鼠的RSH值均同样降低，同时细胞内ROS水平升高或Oxyblot信号增强。个体RSH值与各自的Oxyblot数据以及DCF荧光高度显著负相关。结果证实了ROS在大鼠模型中的衰老和肾上腺素能应激中的作用，并表明血清RSH的降低可作为淋巴细胞氧化应激增强的有效指标。
