Petkovic Igor, Bresgen Nikolaus, Gilardoni Ettore, Regazzoni Luca, Uchida Koji, Aldini Giancarlo, Siems Werner, Eckl Peter
Department of Biosciences, University of Salzburg, Hellbrunnerstrasse 34, 5020 Salzburg, Austria.
Department of Pharmaceutical Sciences, Università degli Studi di Milano, Via Mangiagalli 25, 20133 Milan, Italy.
Antioxidants (Basel). 2020 Feb 11;9(2):150. doi: 10.3390/antiox9020150.
Evidence suggests that the increased production of free radicals and reactive oxygen species lead to cellular aging. One of the consequences is lipid peroxidation generating reactive aldehydic products, such as 4-hydroxynonenal (HNE) that modify proteins and form adducts with DNA bases. To prevent damage by HNE, it is metabolized. The primary metabolic products are the glutathione conjugate (GSH-HNE), the corresponding 4-hydroxynonenoic acid (HNA), and the alcohol 1,4-dihydroxynonene (DHN). Since HNE metabolism can potentially change during in vitro aging, cell cultures of primary human dermal fibroblasts from several donors were cultured until senescence. After different time points up to 30 min of incubation with 5 µM HNE, the extracellular medium was analyzed for metabolites via liquid chromatography coupled with electrospray ionization mass spectrometry (LC/ESI-MS). The metabolites appeared in the extracellular medium 5 min after incubation followed by a time-dependent increase. But, the formation of GSH-HNL and GSH-DHN decreased with increasing in vitro age. As a consequence, the HNE levels in the cells increase and there is more protein modification observed. Furthermore, after 3 h of incubation with 5 µM HNE, younger cells showed less proliferative capacity, while in older cells slight increase in the mitotic index was noticed.
有证据表明，自由基和活性氧的产生增加会导致细胞衰老。其中一个后果是脂质过氧化产生反应性醛类产物，如4-羟基壬烯醛（HNE），它会修饰蛋白质并与DNA碱基形成加合物。为防止HNE造成损伤，它会被代谢。主要代谢产物是谷胱甘肽共轭物（GSH-HNE）、相应的4-羟基壬烯酸（HNA）和醇1,4-二羟基壬烯（DHN）。由于HNE代谢在体外衰老过程中可能会发生变化，因此对来自多个供体的原代人皮肤成纤维细胞进行培养直至衰老。在与5µM HNE孵育长达30分钟的不同时间点后，通过液相色谱与电喷雾电离质谱联用（LC/ESI-MS）分析细胞外培养基中的代谢产物。孵育5分钟后代谢产物出现在细胞外培养基中，随后呈时间依赖性增加。但是，GSH-HNL和GSH-DHN的形成随着体外年龄的增加而减少。结果，细胞内的HNE水平升高，并且观察到更多的蛋白质修饰。此外，在与5µM HNE孵育3小时后，年轻细胞的增殖能力较低，而在老年细胞中观察到有丝分裂指数略有增加。
