Methods of intrinsic and probe fluorescence in assessment of the effect of therapeutic radiation doses in vitro on the albumin molecule

N. Puzan, I. A. Cheshik, V. N. Belyakovsky
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Abstract

   Objective. To study in vitro the effect of therapeutic doses of radiation on the albumin molecule using intrinsic and probe fluorescence methods.   Materials and methods. In order to study radiation-induced changes in serum albumin during in vitro irradiation with therapeutic doses (2 Gy, 40 Gy and 70 Gy), the study was conducted in 2 directions: therapeutic doses of ionizing radiation irradiated a buffer, which was then used to prepare an albumin solution (pre-irradiation of the buffer); therapeutic doses of ionizing radiation irradiated an albumin buffer solution. The presence of structural and functional (conformational) changes in the albumin molecule was judged by changes in the values of intrinsic (λexc=280 nm) and probe (λexc=280nm, λexc=320 nm) fluorescence. Statistical processing of the obtained data was carried out using the program GraphPad Prism 6.0.   Results. Irradiation with therapeutic doses of 2 Gy, 40 Gy and 70 Gy causes conformational changes (a statistically significant decrease in fluorescence intensity) in the albumin molecule, both during preliminary irradiation of the buffer solution used later for the preparation of albumin, and during irradiation of the buffer solution of albumin.   Conclusions. Quantitative changes in the fluorescence intensity, both intrinsic and probe, differ under different modes of albumin irradiation.
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利用本征荧光和探针荧光评估体外治疗辐射剂量对白蛋白分子影响的方法
研究目的使用本征荧光和探针荧光方法,在体外研究治疗剂量辐射对白蛋白分子的影响。 材料和方法。为了研究治疗剂量(2 Gy、40 Gy 和 70 Gy)体外辐照时辐射诱导血清白蛋白的变化,研究分两个方向进行:治疗剂量的电离辐射辐照缓冲液,然后用缓冲液配制白蛋白溶液(缓冲液的预辐照);治疗剂量的电离辐射辐照白蛋白缓冲溶液。根据本征荧光(λexc=280 纳米)和探针荧光(λexc=280 纳米,λexc=320 纳米)值的变化来判断白蛋白分子结构和功能(构象)是否发生变化。使用 GraphPad Prism 6.0 程序对所得数据进行统计处理。 结果在对用于制备白蛋白的缓冲溶液进行初步辐照和对白蛋白缓冲溶液进行辐照时,治疗剂量为 2 Gy、40 Gy 和 70 Gy 的辐照都会导致白蛋白分子构象发生变化(荧光强度在统计学上显著降低)。 结论在不同的白蛋白辐照模式下,荧光强度(包括本征荧光强度和探针荧光强度)的定量变化是不同的。
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