Department of Gerontology and Geriatrics, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
Mech Ageing Dev. 2012 Jul;133(7):498-507. doi: 10.1016/j.mad.2012.06.002. Epub 2012 Jun 18.
Senescence is thought to play an important role in the progressive age-related decline in tissue integrity and concomitant diseases, but not much is known about the complex interplay between upstream regulators and downstream effectors. We profiled whole genome gene expression of non-stressed and rotenone-stressed human fibroblast strains from young and oldest old subjects, and measured senescence associated β-gal activity. Microarray results identified gene sets involved in carbohydrate metabolism, Wnt/β-catenin signaling, the cell cycle, glutamate signaling, RNA-processing and mitochondrial function as being differentially regulated with chronological age. The most significantly differentially regulated mRNA corresponded to the p16 gene. p16 was then investigated using qPCR, Western blotting and immunocytochemistry. In conclusion, we have identified cellular pathways that are differentially expressed between fibroblast strains from young and old subjects.
衰老被认为在组织完整性和伴随疾病的进行性年龄相关下降中起重要作用，但对于上游调节剂和下游效应物之间的复杂相互作用知之甚少。我们对来自年轻和最年长受试者的非应激和鱼藤酮应激的人成纤维细胞株进行了全基因组基因表达谱分析，并测量了衰老相关的β-半乳糖苷酶活性。微阵列结果鉴定了与年龄相关的碳水化合物代谢、Wnt/β-连环蛋白信号、细胞周期、谷氨酸信号、RNA 处理和线粒体功能相关的基因集，这些基因集受到差异调控。差异调节最显著的 mRNA 对应于 p16 基因。然后使用 qPCR、Western blot 和免疫细胞化学法研究了 p16。总之，我们已经鉴定出在来自年轻和老年受试者的成纤维细胞株之间差异表达的细胞途径。
