Effect of Cu2+ on angiogenesis and nucleoli morphology of cultured endothelial cells of the rat cerebral cortex

Stelmashook, E.V.; Genrikhs, E.E.; Alexandrova, O.P.; Strizhkova, A.B.; Lapieva, A.E.; Kapkaeva, M.R.; Isaev, N.K.

Авторы: Stelmashook Elena V., Genrikhs Elizaveta E., Alexandrova Olga P., Strizhkova Anna B., Lapieva Alina E., Kapkaeva Marina R., Isaev Nickolay K.
Журнал: Анналы клинической и экспериментальной неврологии
Том: 20
Номер: 1
Год издания: 2026
Первая страница: 48
Последняя страница: 55
DOI: 10.17816/acen.1437
Аннотация: ntroduction. Wilson–Konovalov disease is associated with impaired intracellular transport of Cu2+, resulting in increased concentrations of un-bound copper in the blood, its accumulation in various organs and tissues, primarily the liver, brain, kidneys, and cornea. The resulting excess Cu2+ ions in the brain leads to altered astrocyte morphology, enlarged microglia, edema of oligodendroglia, reduced neuronal count, and impaired permeability of microcirculatory vessels.The study aimed to determine how Cu2+ excess affects angiogenesis and nucleoli in cultured rat cerebral cortex endothelial cells (ECs).Materials and methods. Copper chloride was added to the culture medium of rat cerebral cortex ECs at concentrations of 50–300 μM for 24 hours. Angiogenesis in cultures was studied using cultured rat brain ECs and the Angiogenesis Assay Kit. Cell viability was assessed using the MTT test, and nucleoli were stained with acridine orange.Results. The effect of Cu2+ on cultured rat cerebral cortex ECs was examined. MTT assay of cultures showed reduced formazan production starting at Cu2+ concentrations of 100 μM in the culture medium, indicating decreased cell viability. At this same concentration, Cu2+-induced impairment of angiogenesis was observed in EC cultures. At higher Cu2+ concentrations (200 μM), surviving cells exhibited a statistically significant increase in nucleolar size to 1.71 ± 0.09 μm2 compared to 1.33 ± 0.07 μm2 in control cultures. Conclusion. Thus, excess copper ions reduce angiogenesis and induce changes in ECs nucleoli, which may represent a universal cellular response associated with cell damage. Keywords: brain; endothelial cells; copper ions; angiogenesis; nucleolusAcknowledgment. The authors express their gratitude to Corresponding Member of the Russian Academy of Sciences A.B. Salmina for her constructive suggestions during the preparation of the work.Ethics approval. The authors confirm compliance with institutional and national standards for laboratory animal use in accor-dance with the Consensus Author Guidelines for Animal Use (IAVES, July 23, 2010). The research protocol was approved by the Local Ethics Committee of the Russian Center of Neurology and Neurosciences (Protocol No. 9-4/23 dated November 29, 2023). Source of funding. The study was supported by the Russian Science Foundation (Grant No. 24-25-00036, https://rscf.ru/project/24-25-00036/).Conflict of interest. The authors declare no apparent or potential conflicts of interest related to the publication of this article.Correspondence address: 5 Obukha Lane, bld 2, Moscow, Russia, 105064. Brain Institute, Russian Center for Neurology and Neuroscience. E-mail: stelmashuk@neurology.ru. Elena V. StelmashookFor citation: Stelmashook E.V., Genrikhs E.E., Alexandrova O.P., Strizhkova A.B., Lapieva A.E., Kapkaeva M.R., Isaev N.K. Effect of Cu2+ on angiogenesis and nucleoli morphology of cultured endothelial cells of the rat cerebral cortex. Annals of Clinical and Experimental Neurology. 2026;20(1):48–55.DOI: https://doi.org/10.17816/ACEN.1437EDN: https://elibrary.ru/NOJGQDReceived 29.10.2025 / Accepted 19.01.2026 / Published 30.03.2026
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