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首页> 外文期刊>Journal of chromatography, B. Analytical technologies in the biomedical and life sciences >A method to quantify intracellular glycation in dermal fibroblasts using liquid chromatography coupled to fluorescence detection – Application to the selection of deglycation compounds of dermatological interest
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A method to quantify intracellular glycation in dermal fibroblasts using liquid chromatography coupled to fluorescence detection – Application to the selection of deglycation compounds of dermatological interest

机译：用液相色谱法耦合到荧光检测中皮肤成纤维细胞内甘油细胞内糖化的方法 - 应用于皮肤病学利益的脱糖化合物

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Glycation is a common non-enzymatic reaction between proteins and sugars, which gives rise in the human body to the formation of advanced glycation end products (AGEs). These modifications impacts both extra and intracellular proteins, leading to cells and tissues dysfunctions. In the skin, accumulation of AGEs leads to aesthetic consequences, wrinkles, dark spots and yellowish skin tone, as it can be seen in diabetic patients. Consequently, there is a growing dermatological interest to find compounds able to eliminate AGEs accumulated in skin.In this context, a method has been developed to detect and quantify intracellular glycation in human dermal fibroblasts. After cultivation of fibroblasts, cell lysates were injected in an HPLC system coupled with a fluorescence detector in by-pass mode. The system allows the simultaneous measurement of global AGEs and particular pentosidine amounts using two sets of wavelengths in a single run of 1?min. The immunocytochemistry approach was used to valid the HPLC analysis data.The method developed was able to quantify changes in global AGEs and pentosidine content in cells in response to glyoxal treatment. Fibroblasts treated with 500?μM of glyoxal for 48?h showed a significant 2.3-fold and 2.6-fold increase in the content of AGEs and pentosidine respectively compared to control cells.As an application, a screening of natural extracts have been done and the method allowed identifying extracts able to significantly reduce the amount of pentosidine in fibroblasts (?32%). These extracts act as deglycation agents of interest in the field of dermatology and cosmetology.
机译：糖化是蛋白质和糖之间的常见非酶促反应，其在人体中产生了先进的糖化末端产物（年龄）。这些修饰会影响额外和细胞内蛋白质，导致细胞和组织功能障碍。在皮肤中，年龄的积累导致审美后果，皱纹，暗点和淡黄色肤色，因为它可以在糖尿病患者中看到。因此，存在不断增长的皮肤病学兴趣，以发现能够消除在皮肤中积累的年龄的化合物。在这种情况下，已经开发了一种方法以检测和量化人类皮肤成纤维细胞中的细胞内糖化。在培养成纤维细胞之后，将细胞裂解物注射在偶联模式下与荧光检测器偶联的HPLC系统中。该系统允许使用两组波长在1·min的波长中同时测量全局年龄和特定的戊胺量。使用免疫细胞化学方法用于有效的HPLC分析数据。作用的方法能够响应于乙二醛处理量量化细胞中全球年龄和戊氨酸含量的变化。用500μl乙二醛处理的成纤维细胞48Ω，分别显着2.3倍和2.6倍的增加与对照细胞相比的年龄和戊糖苷的含量增加。A应用，已经完成了自然提取物的筛选允许鉴定能够显着减少成纤维细胞（β32％）中戊苷的量的提取物。这些提取物充当皮肤病学和美容领域的嗜益腺苷酸。

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