Division of Molecular Dermatology, Department of Dermatology, Paracelsus Private Medical University Salzburg, Salzburg, Austria.
Exp Dermatol. 2010 Oct;19(10):912-8. doi: 10.1111/j.1600-0625.2010.01144.x.
Studies of skin aging are usually performed at the genomic level by investigating differentially regulated genes identified through subtractive hybridization or microarray analyses. In contrast, relatively few studies have investigated changes in protein expression of aged skin using proteomic profiling by two-dimensional (2-D) gel electrophoresis and mass spectrometry, although this approach at the protein level is suggested to reflect more accurately the aging phenotype. We undertook such a proteomic analysis of intrinsic human skin aging by quantifying proteins extracted and fluorescently labeled from sun-protected human foreskin samples pooled from 'young' and 'old' men. In addition, we analyzed these candidate gene products by 1-D and 2-D western blotting to obtain corroborative protein expression data, and by both real-time PCR (RT-PCR) and microarray analyses to confirm expression at the mRNA level. We discovered 30 putative proteins for skin aging, including previously unrecognized, post-translationally regulated candidates such as phosphatidyl-ethanolamine binding protein (PEBP) and carbonic anhydrase 1 (CA1).
皮肤衰老的研究通常在基因组水平上进行，通过调查通过消减杂交或微阵列分析鉴定的差异调节基因。相比之下，使用二维（2-D）凝胶电泳和质谱法通过蛋白质组学分析研究衰老皮肤的蛋白质表达变化的研究相对较少，尽管这种蛋白质水平的方法被认为更准确地反映衰老表型。我们通过从“年轻”和“年老”男性的人保护性包皮样品中提取和荧光标记蛋白质来进行这种内在皮肤衰老的蛋白质组学分析。此外，我们通过 1-D 和 2-D western 印迹分析来获得这些候选基因产物的验证蛋白表达数据，并通过实时 PCR（RT-PCR）和微阵列分析来证实 mRNA 水平的表达。我们发现了 30 种皮肤衰老的潜在蛋白质，包括以前未被识别的、翻译后调节的候选物，如磷酸乙醇胺结合蛋白（PEBP）和碳酸酐酶 1（CA1）。
