Methods of intrinsic and probe fluorescence in assessment of the effect of therapeutic radiation doses in vitro on the albumin molecule
https://doi.org/10.51523/2708-6011.2024-21-1-10
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=280
nm, 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.
Supporting agencies: The work was carried out within the framework of the SPSR “Natural Resources and the Environment” topic “To develop criteria for assessing radiation-induced changes in the tissue of the internal environment based on the analysis of the structure and mechanical properties of the cellular component on in vitro and in vivo models”, reg. № 20210231
About the Authors
N. D. Puzan
Institute of Radiobiology of the National Academy of Sciences of Belarus
Belarus
Belarus
Natallia D. Puzan, Researcher
Gomel
I. A. Cheshik
Institute of Radiobiology of the National Academy of Sciences of Belarus
Belarus
Belarus
Igor A. Cheshik, Candidate of Medical Sciences, Associate Professor, Director
Gomel
V. N. Belyakovsky
Gomel State Medical University
Belarus
Belarus
Vasili N. Belyakovsky, Doctor of Medical Sciences, Professor
Department of Oncology
Gomel
References
1. Korolyuk EV, Korolenko EA, Zhbankov RG, Ivanov AA, Leshchenko V., Ilyich GK, et al. The method of fluorescent sensing in the assessment of albumin transport function. Health-care.1999;1:11-13. (In Russ.).
2. Pushkarev SV, Naumov SA, Vovk SM, Smolyaninov ES, Udut VV. Spectroscopic diagnosis of malignant neoplasms (review). Autometry. 2000;(1):84-93. (In Russ.).
3. Novitsky VV, Stepovaya EA, Batukhtin AV, Goldberg VE, Kolosova MV. Characteristics of the state of erythrocyte membranes in patients with malignant neoplasms according to fluorescence sensing data. The expert bulletin. biol. and medicine. 1999;128(8):226-229. (In Russ.).
4. Cohen BE, Pralle A, Yao X, Swaminath G, Gandhi ChS, Jan YuN, et al. A fluorescent probe designed for studying protein conformational change. Proceedings of the National Academy of Sciences of the USA. 2005;102(4):965-970. doi: 10.1073/pnas.0409469102
5. Kirilova EM, Kalnina I, Zvagule T, Gabruseva N, Kurjane N, Solomenikova II. Fluorescent study of human blood plasma albumin alterations induced by ionizing radiation. Journal of Fluorescence. 2011:21(3):923-927. doi: 10.1007/s10895-010-0608-2
6. Gryzynov YuA., Dobretsov GE. Blood serum albumin in clinical medicine. Moscow; 1994. (In Russ.).
7. Ota C, Takano K. Spectroscopic analysis of protein crowded environments using the charge-transfer fluorescence probe ANS. ChemPhysChem. 2019;20(11):1456-1466. doi: 10.1002/cphc.201900226
8. Puzan ND, Suslova AA. The effect of therapeutic doses of ionizing radiation on the conformational state of serum albumin. In: Radiobiology: challenges of the XXI century : materials of the International scientific conference dedicated to the 30<sup>th</sup> anniversary of the Institute of Radiobiology; 2017, September 27-30; Gomel. Gomel: Institute of Radiobiology of the National Academy of Sciences of Belarus; 2017. pp. 152-154. [date of access 2023 July 13]. Available from: https://drive.google.com/file/d/0B6K-Pvik5WACaG5SeEtFUF91X0U/view?resource-key=0-I0RVvZZNMlkKUU_RJdlD5A (In Russ.).
9. Gudkov SV, Garmash SA, Karp OE, Smirnova VS, Chernikov AV, Bruskov VI. Long-lived amino acid radicals induced by X-ray radiation are a source of hydrogen peroxide formation in an aqueous medium. Biophysics. 2010;55(4):588-593. (In Russ.).
10. Rubtsova EV. Investigation of the topology of hydrate shells of proteins. [Electronic resource]. The website of the Russian State Library. [date of access 2024 January 12]. Available from: https://viewer.rsl.ru/ru/rsl01005564868?page=1&rotate=0&theme=white (In Russ.)
11. Burlakova E.B., Kondratov A.A., Maltseva E.L. The effect of ultra-low doses of biologically active substances and low-intensity physical factors. Chemical physics. 2003; 22(2):21-40. (In Russ.).
Review
For citations:
Puzan N.D., Cheshik I.A., Belyakovsky V.N. Methods of intrinsic and probe fluorescence in assessment of the effect of therapeutic radiation doses in vitro on the albumin molecule. Health and Ecology Issues. 2024;21(1):81-88. (In Russ.) https://doi.org/10.51523/2708-6011.2024-21-1-10
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