Evans Nicholas D, Gnudi Luigi, Rolinski Olaf J, Birch David J S, Pickup John C
Metabolic Unit, Guy's, King's and St Thomas's School of Medicine, Guy's Hospital, London, United Kingdom.
Diabetes Technol Ther. 2003;5(5):807-16. doi: 10.1089/152091503322527012.
The aim of this study was to develop an in vitro cell-culture model of skin-component cells to test the hypothesis that glucose can be monitored non-invasively by measuring NAD(P)H-related fluorescence changes in tissues. 3T3-L1 fibroblasts and adipocytes were grown in culture, and the response to added glucose was assessed by changes in steady-state autofluorescence at 400-500 nm [excitation at 340 nm, an index of NAD(P)H]. We also studied glucose-related fluorescence changes in cells stained with the mitochondrial marker, rhodamine-123. Fibroblasts and adipocytes showed glucose-dependent increases in autofluorescence with both short- and long-term exposure. Spectral properties indicated that the fluorescence was due to NAD(P)H production. With 5-h exposure to glucose, the maximal response was at 10-15 mmol/L glucose. Cells stained with the fluorescent mitochondrial marker, rhodamine-123, showed an immediate and marked decrease in fluorescence when exposed to glucose. We conclude that glucose can be sensed non-invasively by cellular fluorescence changes in fibroblasts and adipocytes. This is a model for the further exploration of fluorescence-based non-invasive metabolic monitoring in human diabetes.
本研究的目的是建立一种皮肤组成细胞的体外细胞培养模型，以验证通过测量组织中与NAD(P)H相关的荧光变化可进行无创血糖监测这一假设。将3T3-L1成纤维细胞和脂肪细胞进行培养，通过在400 - 500 nm处稳态自发荧光的变化（激发波长为340 nm，NAD(P)H的一个指标）评估对添加葡萄糖的反应。我们还研究了用线粒体标记物罗丹明-123染色的细胞中与葡萄糖相关的荧光变化。成纤维细胞和脂肪细胞在短期和长期暴露下均显示出自发荧光随葡萄糖浓度增加。光谱特性表明荧光是由NAD(P)H产生所致。暴露于葡萄糖5小时后，最大反应出现在葡萄糖浓度为10 - 15 mmol/L时。用荧光线粒体标记物罗丹明-123染色的细胞在暴露于葡萄糖时显示出荧光立即且显著降低。我们得出结论，通过成纤维细胞和脂肪细胞中的细胞荧光变化可无创检测葡萄糖。这是进一步探索基于荧光的人类糖尿病无创代谢监测的一个模型。
