Istituto di Biomembrane e Bioenergetica, CNR, Bari, Italy.
Mech Ageing Dev. 2011 Oct;132(10):488-95. doi: 10.1016/j.mad.2011.07.009. Epub 2011 Aug 9.
To find out whether and how deficit of cellular energy supply from mitochondria to cytosol occurs in aging and hypertension, we used mitochondria isolated from 5 to 72 week-old heart left ventricle of either normotensive (WKY) or spontaneous hypertensive (SH) rats as a model system. Measurements were made of the rate of ATP appearance outside mitochondria, due to externally added ADP, as an increase of NADPH absorbance which occurs when ATP is produced in the presence of glucose, hexokinase and glucose-6-phosphate dehydrogenase. Such a rate proved to mirror the function of the adenine nucleotide translocator (ANT) rather than other processes linked to the both oxidative and substrate level phosphorylation. The changes in both Ki for atractyloside and Km for ADP suggest the occurrence of modification of the ANT conformation during aging in which the ANT Vmax was found to decrease in normotensive but to increase under spontaneously hypertension in 24 week-old rats with a subsequent decrease in both cases. ANT function, as investigated in the ADP physiological range (20-60μM), is expected to decrease in normotensive, but to increase in hypertensive rats up to 48 weeks. Later a decrease in the ATP rate of export outside mitochondria should occur in both cases.
为了探究线粒体到细胞质的细胞能量供应不足是否以及如何在衰老和高血压中发生,我们使用了从 5 至 72 周龄的正常血压(WKY)或自发性高血压(SH)大鼠的左心室心肌分离的线粒体作为模型系统。测量由于外加 ADP 而在线粒体外部出现的 ATP 生成速率,这是由于在存在葡萄糖、己糖激酶和葡萄糖-6-磷酸脱氢酶的情况下产生 ATP 时 NADPH 吸光度的增加。该速率被证明反映了嘌呤核苷酸转运蛋白(ANT)的功能,而不是与氧化磷酸化和底物水平磷酸化都相关的其他过程。阿曲霉素的 Ki 和 ADP 的 Km 的变化表明,在衰老过程中,ANT 构象发生了修饰,在 24 周龄的正常血压大鼠中发现 ANT Vmax 降低,而在自发性高血压大鼠中增加,随后两种情况下均降低。在 ADP 的生理范围内(20-60μM)研究的 ANT 功能预计在正常血压大鼠中降低,但在高血压大鼠中增加,直至 48 周。之后,在线粒体外部的 ATP 输出速率应该在两种情况下都降低。