Glucose-dependent changes in NAD(P)H-related fluorescence lifetime of adipocytes and fibroblasts in vitro: Potential for non-invasive glucose sensing in diabetes mellitus
Glucose-dependent changes in NAD(P)H-related fluorescence lifetime of adipocytes and fibroblasts in vitro: Potential for non-invasive glucose sensing in diabetes mellitus
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DOI：
10.1016/j.jphotobiol.2005.04.001
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发表时间：
2005-08-01
影响因子：
5.4
通讯作者：
Pickup, JC
中科院分区：
文献类型：
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作者：
Evans, ND;Gnudi, L;Pickup, JC
本研究的目的是测试的假设，葡萄糖可以通过在体外细胞培养模型中测量NAD（P）H相关的荧光寿命的细胞进行非侵入性监测。通过时间相关单光子计数（激发370 nm，发射420-480 nm）测量3 T3-L1脂肪细胞中的自体荧光衰减函数。游离NADH有两个指数衰减，但细胞自发荧光最适合三个指数衰减。添加30 mM葡萄糖导致自发荧光强度增加29%，平均寿命显著缩短（从7.23到6.73 ns），并且以两个较长寿命组分为代价增加了短寿命组分的相对振幅和分数强度。鱼藤酮也有类似的效果，鱼藤酮是一种使线粒体NADH最大化的药物。用荧光线粒体标记物罗丹明123染色的3 T3-L1成纤维细胞在暴露于30 mM葡萄糖时显示出16%的荧光强度猝灭，并且以较长寿命组分为代价增加了短寿命的相对振幅和分数强度。我们的结论是，虽然影响的大小是相对较小的，葡萄糖可以测量非侵入性的细胞通过监测细胞自发荧光或线粒体代谢的染料标记物的寿命的变化。因此，荧光强度和寿命传感的进一步研究和发展表明，可能的非侵入性代谢监测在人类糖尿病。(c)2005 Elsevier B. V.保留所有权利。
The aim of this study was to test the hypothesis that glucose can be monitored non-invasively by measuring NAD(P)H-related fluorescence lifetime of cells in an in vitro cell culture model. Autofluorescence decay functions were measured in 3T3-L1 adipocytes by time-correlated single-photon counting (excitation 370 nm, emission 420-480 nm). Free NADH had a two-exponential decay but cell autofluorescence fitted best to a three-exponential decay. Addition of 30 mM glucose caused a 29% increase in autofluorescence intensity, a significantly shortened mean lifetime (from 7.23 to 6.73 ns), and an increase in the relative amplitude and fractional intensity of the short-lifetime component at the expense of the two longer-lifetime components. Similar effects were seen with rotenone, an agent that maximizes mitochondrial NADH. 3T3-L1 fibroblasts stained with the fluorescent mitochondrial marker, rhodamine 123 showed a 16% quenching of fluorescence intensity when exposed to 30 mm glucose, and an increase in the relative amplitude and fractional intensity of the short lifetime at the expense of the longer lifetime component. We conclude that, though the effect size is relatively small, glucose can be measured non-invasively in cells by monitoring changes in the lifetimes of cell autofluorescence or of a dye marker of mitochondrial metabolism. Further investigation and development of fluorescence intensity and lifetime sensing is therefore indicated for possible non-invasive metabolic monitoring in human diabetes. (c) 2005 Elsevier B.V. All rights reserved.