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

摘要

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.Auto fluorescence 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 rote-none,an agent that maximizes mitochondrial NADH.3T3-L1 fibroblasts stained with the fluorescent mitochondrial marker,rho-damine 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 auto fluorescence 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.