NADH/FAD溶液中时变光学氧化还原比的光谱研究,The Journal of Physical Chemistry B

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NADH/FAD溶液中时变光学氧化还原比的光谱研究
The Journal of Physical Chemistry B ( IF 2.8 ) Pub Date : 2022-11-18 , DOI: 10.1021/acs.jpcb.2c05292
Soo Yeong Lim ₁ , Jin Il Jang ₁ , Hongman Yoon ₂ , Hyung Min Kim ₁

Affiliation

自发荧光成像已被广泛用作体内和离体组织的高级无创诊断。光学氧化还原比 (ORR) 被定义为还原型尼古丁腺嘌呤二核苷酸 (NADH) 和氧化黄素腺嘌呤二核苷酸 (FAD) 之间的荧光强度比，已被强烈用作诊断参数，因为 NADH 和 FAD 起着重要作用在能量和呼吸代谢中作为辅酶。ORR 方法已在包括乳腺癌、宫颈癌和口腔癌在内的癌症诊断中提供了成功的评估；关于两个分子之间的光学和化学干扰导致 ORR 值发生变化的研究很少。在这项研究中，我们研究了溶解在三（羟甲基）氨基甲烷、磷酸盐缓冲液中的 NADH/FAD 混合物的 ORR 值变化，和去离子水环境。制备各种浓度比的体外溶液，并从时间序列的荧光和吸收光谱中获得实验和理论 ORR 值。基于光谱分析，我们得出结论，内部过滤效应导致 FAD 荧光在溶解后立即降低，氧化还原与氧化反应相结合导致 NADH 荧光和 FAD 发射随时间变化降低。

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Spectroscopic Study of Time-Varying Optical Redox Ratio in NADH/FAD Solution

Autofluorescence imaging has been widely applied as advanced noninvasive diagnostics for in vivo and ex vivo tissues. The optical redox ratio (ORR), which is defined as the fluorescence intensity ratio between reduced nicotine adenine dinucleotide (NADH) and oxidized flavin adenine dinucleotide (FAD), has been used as a diagnostic parameter strongly, because NADH and FAD play an important role in energetic and respiratory metabolism as coenzymes. The ORR method has provided successful assessment in cancer diagnosis including breast, cervical, and oral cancer; few studies have been reported about optical and chemical interference between two molecules resulting in a change in ORR values. In this study, we investigated the variations in ORR values of NADH/FAD mixtures dissolved in tris(hydroxymethyl)aminomethane, phosphate buffer, and deionized water environments. In vitro solutions were prepared in various concentration ratios and the experimental and theoretical ORR values were obtained from fluorescence and absorption spectra in time series. Based on the spectroscopic analysis, we concluded that the inner filter effect causes an instant decrease in FAD fluorescence just after dissolution and that the oxidation–reduction coupled with oxygenation reaction results in time-varying decreases in NADH fluorescence and FAD emission.

更新日期：2022-11-18

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