Archives2022Vol. 62, № 2pp. 171–179

Article

Analysis of Markers of Oxidative Damage of Neurons and Neuroinflammation Late after γ-Irradiation of the Mice Head at Different Doses

E. Yu. Moskaleva, A. V. Rodina, Yu. P. Semochkina, O. V. Vysotskaya

National Research Center “Kurchatov Institute”, Moscow, Russia

Abstract

Reactive oxygen species (ROS), formed under the action of ionizing radiation and of inflammation factors, react with lipids, lead to the formation of toxic metabolites, one of the most active among which is 4-hydroxy-2-nonenal (4-HNE). The aim of this work is to study the number of neurons containing 4-HNE-modified proteins 2 months after γ-irradiation of the head at doses of 2, 8, and 20 Gy, and to determine the development of neuroinflammation at this period. Mouse brain cells were isolated by centrifugation through Percoll solution after brain tissue treatment with accutase. The number of neurons and neurons containing 4-HNEmodified proteins, as well as the number of cells of resting and activated microglia were analyzed using flow cytometry, the level of the proinflammatory cytokine TNFα gene expression in the hippocampus was analyzed using RT-PCR. It was shown that in the brains of control mice, 14.3 ± 2.3% of neurons contain 4-HNE-modified proteins. Two months after irradiation of the head of mice at doses of 8 and 20 Gy the number of 4-HNE+ neurons increased to 23.1 ± 2.6% and 34.8 ± 4.9% (p < 0.05). An increase in the proportion of activated microglia in isolated brain cells and in the level of TNFα gene expression in the hippocampus at this period indicates the development of neuroinflammation. Thus, in addition to direct damage during irradiation, neurons are exposed to ROS and toxic metabolites of lipid peroxidation, which are formed both at the time of irradiation and during the neuroinflammation, which develops late after radiation exposure.

Keywords

4-hydroxy-nonenal, lipid peroxidation, neurons, gamma radiation, local irradiation, neuroinflammation, microglia, TNFα, brain, mouse

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