VitroScreen In Vitro Research Laboratories, Milan, Italy.
Professional Dietetics, Milan, Italy.
J Biol Regul Homeost Agents. 2018 May-Jun;32(3):517-527.
The aim of this screening study was to evaluate the efficacy of different proprietary mixtures of amino acid and hyaluronic acid (HA) in stimulating the production of extracellular matrix (ECM) components, particularly the neo-synthesis of elastin, and in promoting a more efficient deposition of elastic fibres (elastogenesis), while at the same time maintaining the stimulation of collagen. The study has allowed identification of the optimal ratios between the amino acids (AA) for the production of collagen and elastin. Human primary dermal fibroblasts from a 44-year-old female donor were used as a test system in an experimental design based on the evaluation of the expression of relevant ECM genes using a transcriptomic dynamic approach. The expression of ECM genes was evaluated by RTqPCR from 24 to 120 hours in the presence of the test items. Moreover, the production of ECM proteins was verified by Western blot analysis after a 120 h treatment period. In addition to elastin, collagen IV, a fundamental structural component of the basal lamina responsible for epithelial and connective tissue anchoring, was analysed as potential target for the modulation of ECM protein production by human fibroblast. The first phase of the study demonstrated that alanine and valine are essential to promote production of elastin, of which they are important constituents. The second phase of the study, which was conducted to clarify the interactions between the different clusters of AA, demonstrated that it is necessary to choose a mixture that contains specific amounts of amino acids of both proteins, collagen and elastin, to give a significant response and a significant production of both. This also proves the existence of a ratio between the 2 clusters (AA elastin/AA collagen) that guarantees an adequate and balanced response to gene expression and production by fibroblasts, collagen and elastin. The study has allowed identification of the optimal ratios between the AA for the production of collagen and elastin.
本筛选研究旨在评估不同专利的氨基酸和透明质酸 (HA) 混合物在刺激细胞外基质 (ECM) 成分产生的功效,特别是弹性蛋白的新合成,并促进更有效地沉积弹性纤维(弹性发生),同时保持对胶原蛋白的刺激。该研究已经确定了产生胶原蛋白和弹性蛋白的最佳氨基酸 (AA) 比例。本研究以 44 岁女性供体的人原代真皮成纤维细胞为测试系统,基于使用转录组动态方法评估相关 ECM 基因表达的实验设计。在存在测试项目的情况下,通过 RTqPCR 从 24 小时到 120 小时评估 ECM 基因的表达。此外,在 120 小时处理期后通过 Western blot 分析验证 ECM 蛋白的产生。除了弹性蛋白,胶原蛋白 IV 作为基底膜的基本结构成分,负责上皮和结缔组织锚定,也被分析为调节人成纤维细胞 ECM 蛋白产生的潜在靶标。研究的第一阶段表明,丙氨酸和缬氨酸对于促进弹性蛋白的产生至关重要,它们是弹性蛋白的重要组成部分。研究的第二阶段旨在澄清不同 AA 簇之间的相互作用,表明有必要选择一种混合物,其中包含两种蛋白质、胶原蛋白和弹性蛋白的特定量的氨基酸,以产生显著的反应和大量的胶原蛋白和弹性蛋白的产生。这也证明了两种簇 (AA 弹性蛋白/AA 胶原蛋白) 之间存在比例关系,可确保成纤维细胞、胶原蛋白和弹性蛋白的基因表达和产生得到适当和平衡的反应。该研究已经确定了产生胶原蛋白和弹性蛋白的最佳氨基酸 (AA) 比例。