Department of Clinical Lab Sciences, College of Applied Medical Sciences, King Saud University, Riyadh, Saudi Arabia.
Ann Nutr Metab. 2011;59(2-4):127-38. doi: 10.1159/000334069. Epub 2011 Dec 2.
To investigate the effect of senescence and selenium supplementation on glutathione peroxidase (cGPx) and catalase (CAT) activities, and concurrent hydrogen peroxide (H(2)O(2)) generation in subcultured human fibroblasts.
cGPx and CAT activities and H(2)O(2) levels were assayed in presenescent passage 5 and 10 cells, and in senescent passage 20, 25, 30 and 35 cells cultured in routine medium (MEM1) and supplemented media MEM2 and MEM3 containing normal and triple human plasma levels of Se, respectively. Senescent cells were identified by studying their growth and replication states, and by monitoring their activity of key glucose and glycogen degradative enzymes.
cGPx activity showed moderate increases in senescent cells at passages 20-35 subcultured in MEM1 or MEM2. This activity underwent highly significant progressive increases in the same senescent cells subcultured in MEM3. In contrast, CAT activity showed progressive, highly significant increases in senescent cells at passages 20-35 regardless of the culture medium type. Concurrent H(2)O(2) generation was significantly increased in passage 15-25 cells and peaked to higher levels in passage 30 and 35 cells cultured in MEM1 or MEM2. These rates, however, were significantly reduced in senescent passage 20-35 cells cultured in MEM3.
The highest cGPx activity and coupled lower H(2)O(2) generation were achieved in senescent cells cultured in MEM3.
研究衰老和硒补充对谷胱甘肽过氧化物酶 (cGPx) 和过氧化氢酶 (CAT) 活性的影响,以及在传代培养的人成纤维细胞中同时产生的过氧化氢 (H2O2)。
在常规培养基 (MEM1) 和补充培养基 MEM2 和 MEM3 中培养的未衰老传代 5 和 10 代细胞以及衰老传代 20、25、30 和 35 代细胞中测定 cGPx 和 CAT 活性和 H2O2 水平。衰老细胞通过研究其生长和复制状态以及监测其关键葡萄糖和糖原降解酶的活性来鉴定。
在 MEM1 或 MEM2 中培养的衰老细胞在传代 20-35 代时 cGPx 活性适度增加。在 MEM3 中培养的相同衰老细胞中,该活性经历了显著的渐进性增加。相比之下,CAT 活性在传代 20-35 代的衰老细胞中呈渐进性、显著增加,无论培养基类型如何。在 MEM1 或 MEM2 中培养的传代 15-25 代细胞中,H2O2 的产生显著增加,并在传代 30 和 35 代细胞中达到更高水平。然而,在 MEM3 中培养的衰老传代 20-35 代细胞中,这些速率显著降低。
在 MEM3 中培养的衰老细胞中实现了最高的 cGPx 活性和耦合更低的 H2O2 生成。