Department of Gerontology and Geriatrics, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
Aging Cell. 2009 Sep;8(5):595-603. doi: 10.1111/j.1474-9726.2009.00506.x. Epub 2009 Aug 4.
Unlike various model organisms, cellular responses to stress have not been related to human longevity. We investigated cellular responses to stress in skin fibroblasts that were isolated from young and very old subjects, and from offspring of nonagenarian siblings and their partners, representatives of the general population. Fibroblasts were exposed to rotenone and hyperglycemia and assessed for senescence-associated beta-galactosidase (SA-beta-gal) activity by flow cytometry. Apoptosis/cell death was measured with the Annexin-V/PI assay and cell-cycle analysis (Sub-G1 content) and growth potential was determined by the colony formation assay. Compared with fibroblasts from young subjects, baseline SA-beta-gal activity was higher in fibroblasts from old subjects (P = 0.004) as were stress-induced increases (rotenone: P < 0.001, hyperglycemia: P = 0.027). For measures of apoptosis/cell death, fibroblasts from old subjects showed higher baseline levels (Annexin V+/PI+ cells: P = 0.040, Sub-G1: P = 0.014) and lower stress-induced increases (Sub-G1: P = 0.018) than fibroblasts from young subjects. Numbers and total size of colonies under nonstressed conditions were higher for fibroblasts from young subjects (P = 0.017 and 0.006, respectively). Baseline levels of SA-beta-gal activity and apoptosis/cell death were not different between fibroblasts from offspring and partner. Stress-induced increases were lower for SA-beta-gal activity (rotenone: P = 0.064, hyperglycemia: P < 0.001) and higher for apoptosis/cell death (Annexin V+/PI- cells: P = 0.041, Annexin V+/PI+ cells: P = 0.008). Numbers and total size of colonies under nonstressed conditions were higher for fibroblasts from offspring (P = 0.001 and 0.024, respectively) whereas rotenone-induced decreases were lower (P = 0.008 and 0.004, respectively). These data provide strong support for the hypothesis that in vitro cellular responses to stress reflect the propensity for human longevity.
与各种模式生物不同，细胞对压力的反应与人类的长寿并没有关系。我们研究了从年轻和非常年老的供体、非百岁老人的兄弟姐妹及其配偶的后代中分离出来的皮肤成纤维细胞对压力的反应，这些供体代表了一般人群。通过流式细胞术评估了成纤维细胞对鱼藤酮和高血糖的衰老相关β-半乳糖苷酶（SA-β-gal）活性。通过 Annexin-V/PI 测定和细胞周期分析（Sub-G1 含量）测量细胞凋亡/死亡，通过集落形成测定法确定生长潜能。与年轻供体的成纤维细胞相比，老年供体的成纤维细胞的基础 SA-β-gal 活性更高（P = 0.004），应激诱导的增加也是如此（鱼藤酮：P < 0.001，高血糖：P = 0.027）。对于细胞凋亡/死亡的测量，与年轻供体的成纤维细胞相比，老年供体的成纤维细胞的基础水平更高（Annexin V+/PI+细胞：P = 0.040，Sub-G1：P = 0.014），应激诱导的增加更低（Sub-G1：P = 0.018）。在非应激条件下，成纤维细胞的集落数量和总大小更高（P = 0.017 和 0.006）。与年轻供体的成纤维细胞相比，后代及其配偶的成纤维细胞的基础 SA-β-gal 活性和细胞凋亡/死亡水平没有差异。应激诱导的增加，SA-β-gal 活性较低（鱼藤酮：P = 0.064，高血糖：P < 0.001），细胞凋亡/死亡较高（Annexin V+/PI-细胞：P = 0.041，Annexin V+/PI+细胞：P = 0.008）。在非应激条件下，后代的成纤维细胞的集落数量和总大小更高（P = 0.001 和 0.024），而鱼藤酮诱导的减少较低（P = 0.008 和 0.004）。这些数据强烈支持这样的假设，即体外细胞对压力的反应反映了人类长寿的倾向。
