Natural moisturizing factor components in the stratum corneum as biomarkers of filaggrin genotype: evaluation of minimally invasive methods
Natural moisturizing factor components in the stratum corneum as biomarkers of filaggrin genotype: evaluation of minimally invasive methods
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DOI：
10.1111/j.1365-2133.2009.09342.x
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发表时间：
2009-11-01
影响因子：
10.3
通讯作者：
Bos, J. D.
中科院分区：
文献类型：
--
作者：
Kezic, S.;Kammeyer, A.;Bos, J. D.
聚丝蛋白基因（FLG）功能缺失突变的携带者会降低角质层中天然保湿因子（NMF）的水平。因此，角质层中聚丝蛋白分解形成的NMF成分的浓度可能可用作FLG基因型的生物标志物。目的探讨不同取样方法测定角质层中2-吡啶酮-5-羧酸（PCA）和尿酸（UCA）两种NMF成分作为FLG基因型生物标志物的可行性。方法采用胶带剥离法和含氢氧化钾（KOH）皮肤贴提取法分别对角质层的PCA和UCA进行取样。采用高效液相色谱法测定PCA和UCA的浓度。该研究包括11个FLG突变携带者和10个最常见的FLG突变（R501X和2282del4）的野生型个体。结果条带剥离法提取的PCA中FLG基因型差异最显著。纯合子（或复合杂合子）、杂合子和野生型基因型的PCA均值分别为0.18、0.50和1.64 mmol g（-1）蛋白（纯合子vs杂合子P < 0.005；杂合子vs野生型P < 0.0001）。与KOH贴片相比，胶带贴片技术显示出更小的受试者内部变化，特别是当胶带上的UCA和PCA浓度与蛋白质量归一化时。结论胶带剥离采集的角质层PCA浓度可作为FLG基因型的生物标志物。
Background The carriers of loss-of-function mutations in the filaggrin gene (FLG) have reduced levels of natural moisturizing factor (NMF) in the stratum corneum. The concentration of NMF components which are formed by filaggrin protein breakdown in the stratum corneum might therefore be useful as a biomarker of the FLG genotype.Objectives To investigate the feasibility of different sampling methods for the determination of two NMF components, 2-pyrrolidone-5-carboxylic acid (PCA) and urocanic acid (UCA), in the stratum corneum as biomarkers for the FLG genotype.Methods PCA and UCA from the stratum corneum were sampled by using a tape stripping technique and an extraction technique using skin patches containing potassium hydroxide (KOH). The concentrations of PCA and UCA were measured by high-performance liquid chromatography. Eleven carriers of an FLG mutation and 10 individuals wild type for the two most common FLG mutations (R501X and 2282del4) were included in the study.Results The most significant difference between the FLG genotypes was found for PCA sampled by the tape stripping technique. The mean values of PCA obtained by the tape stripping technique were, respectively, 0 18, 0 50 and 1 64 mmol g(-1) protein in homozygous (or compound heterozygous), heterozygous and wild-type genotypes (P < 0 005 homozygous vs. heterozygous; P < 0 0001 heterozygous vs. wild type). The tape stripping technique showed less intrasubject variation compared with the KOH patches, in particular when the concentrations of UCA and PCA on the tape strips were normalized for protein amount.Conclusions The concentration of PCA in the stratum corneum collected by tape stripping showed it to be a feasible biomarker of the FLG genotype.