Early neuroprotection: effects on p53 and annexin A5 protein expression in the basal magnocellular nucleus of rats with colchicine-induced injury

Authors

DOI:

https://doi.org/10.14739/2310-1210.2026.3.352377

Keywords:

neuroprotection, neurodegeneration, apoptosis, p53, Annexin A5, brain, neurons, citicoline, thiocetam, HSF-1

Abstract

Aim. To determine the expression patterns of p53 and annexin A5 in the cells of the basal magnocellular nucleus (BMN) of the rat brain following early neuroprotective intervention with citicoline, thiocetam, and recombinant HSF-1 in a rat model of colchicine-induced neurodegeneration.

Materials and methods. The study involved 50 male Wistar rats divided into five groups (n = 10 each). Group 1 (control) received an intracerebroventricular (ICV) injection of 0.9 % NaCl solution. Animals in groups 2–5 received ICV colchicine. Group 2 served as the positive control (untreated pathology model). Groups 3–5 received citicoline, thiocetam, and HSF-1, respectively, for 14 consecutive days beginning the day after surgery. Subsequently, all rats were euthanized under thiopental sodium anesthesia, and the brains were harvested for histochemical and immunofluorescent analyses.

Results. ICV colchicine caused loss of the neuronal component and an increase in glial cell number in the BMN. A two-week course of neuroprotective correction prevented these changes. Colchicine-induced neurodegeneration was also associated with an elevated number of p53- and Annexin A5-immunopositive cells in the BMN. Notably, administration of citicoline and HSF-1 resulted in a further pronounced increase in p53-positive neurons (by 60–70 % relative to the pathology group), whereas thiocetam caused a significantly increased number of annexin A5-immunopositive cells. Concurrently, all types of pathogenetic correction normalized the Corrected Total Cell Fluorescence (CTCF) of annexin A5+ cells to control levels.

Conclusions. In the BMN of experimental animals on day 14 post-ICV colchicine injection, a decrease in neuronal density and an increase in glial density were observed compared to sham-operated rats. In contrast, in the groups receiving early administration of citicoline, thiocetam, or recombinant HSF-1, neuronal and glial density parameters remained at levels comparable to control animals. The highest CTCF values for annexin A5-positive cells were recorded in the colchicine group compared to both sham-operated and correction groups, with no statistically significant differences among the latter. At the same time, p53 CTCF values in the BMN across all experimental groups showed no statistically significant differences compared to the control or each other. While the total counts of p53- and annexin A5-positive cells in the colchicine group significantly exceeded those in sham-operated animals, this parameter in the correction groups varied according to the specific pharmacological agent used.

Author Biography

M. V. Danukalo, Zaporizhzhia State Medical and Pharmaceutical University

MD, PhD, Associate Professor of the Department of Pathological Physiology with the Course of Normal Physiology

References

  1. Chen YS, Shu K, Kang HC. Deep brain stimulation in Alzheimer’s disease: Targeting the nucleus basalis of Meynert. J Alzheimers Dis. 2021;80(1):53-70. doi: https://doi.org/10.3233/JAD-201141
    | |
  2. Tiernan CT, Mufson EJ, Kanaan NM, Counts SE. Tau oligomer pathology in nucleus basalis neurons during the progression of Alzheimer disease. J Neuropathol Exp Neurol. 2018;77(3):246-59. doi: https://doi.org/10.1093/jnen/nlx120
    | |
  3. Nazmuddin M, Philippens IH, van Laar T. Electrical stimulation of the nucleus basalis of Meynert: a systematic review of preclinical and clinical data. Sci Rep. 2021;11(1):11751. doi: https://doi.org/10.1038/s41598-021-91391-0
    | |
  4. Rapaka D, Adiukwu PC, Bitra VR. Experimentally induced animal models for cognitive dysfunction and Alzheimer’s disease. MethodsX. 2022;9:101933. doi: https://doi.org/10.1016/j.mex.2022.101933
    | |
  5. Kumar A, Seghal N, Naidu PS, Padi SS, Goyal R. Colchicines-induced neurotoxicity as an animal model of sporadic dementia of Alzheimer's type. Pharmacol Rep. 2007;59(3):274-83.
    | |
  6. Kaji AA, Torii M, Ishii S. Caspase-3 inhibition toward perinatal protection of the developing brain from environmental stress. Dev Neurosci. 2023;45(2):66-75. doi: https://doi.org/10.1159/000529125
    | |
  7. Danukalo MV, Kolesnyk YM. The specificity of iNOS expression indicators in the basal magnocellular nucleus of rats under early pathogenetic correction in experimental neurodestruction. Zaporozhye Medical Journal. 2024;26(5):379-86. doi: https://doi.org/10.14739/2310-1210.2024.5.309732
    |
  8. Belenichev IF, Aliyeva OG, Kamyshnyi AM. Long-term results of pharmacological correction of iNOS, eNOS, nNOS mRNA expression disorders in rat hippocampus after chronic prenatal hypoxia. Biological Markers in Fundamental and Clinical Medicine. 2019;3(2):6-15. doi: https://doi.org/10.29256/v.03.02.2019.escbm01-06
  9. Demchenko AV, Belenichev ІF. [Estimation of thiocetam influence on glutathione link of thiol disulfde cerebral system in conditions of experimental chronic ischemia]. Pathologia. 2015;(2):101-5. Ukrainian. doi: https://doi.org/10.14739/2310-1237.2015.2.51154
  10. Burlaka BS, Belenichev IF, Ryzhenko OI, Ryzhenko VP, Aliyeva OG, Makyeyeva LV, et al. The effect of intranasal administration of an IL-1b antagonist (RAIL) on the state of the nitroxydergic system of the brain during modeling of acute cerebrovascular accident. Pharmacia. 2021;68(3):665-70. doi: https://doi.org/10.3897/pharmacia.68.e71243
    |
  11. Belenichev IF, Aliyeva OG, Gunina LM, Bukhtiyarova NV. [Evaluation of the efficiency of the neuroprotective drugs after prenatal hypoxia]. Fiziolohichnyi zhurnal. 2023;69(1):43-53. Ukrainian. doi: https://doi.org/10.15407/fz69.01.043
  12. Danukalo MV, Kolesnyk YM, Hancheva OV. Contemporary comprehensive approaches to assessing the effectiveness of experimental model of neurodegenerative disorders with cognitive status changes. Mod Med Technol. 2023;(4):51-8. doi: https://doi.org/10.34287/MMT.4(59).2023.7
    |
  13. Paxinos G, Watson C. The rat brain in stereotaxic coordinates. 7th ed. London: Elsevier Academic Press; 2018.
  14. SCBT. Santa Cruz biotechnology [Internet]. SCBT. [cited 2026 Mar 14]. Available from: https://www.scbt.com/resources/protocols/immunofluorescence-cell-staining
  15. Liang Y, Li S, Wen C, Zhang Y, Guo Q, Wang H, et al. Intrastriatal injection of colchicine induces striatonigral degeneration in mice. J Neurochem. 2008;106(4):1815-27. doi: https://doi.org/10.1111/j.1471-4159.2008.05526.x
    | |
  16. Belenichev IF, Cherniy V, Nahorna E, Pavlov S, Cherniy T, Bukhtiyarova N, et al. [Neuroprotection and neuroplasticity]. Kyiv: Logos; 2015. Ukrainian.
  17. Almeida A, Sánchez-Morán I, Rodríguez C. Mitochondrial-nuclear p53 trafficking controls neuronal susceptibility in stroke. IUBMB Life. 2021;73(3):582-91. doi: https://doi.org/10.1002/iub.2453
    | |
  18. Gao Y, Zhang H, Wang J, Li F, Li X, Li T, et al. Annexin A5 ameliorates traumatic brain injury-induced neuroinflammation and neuronal ferroptosis by modulating the NF-ĸB/HMGB1 and Nrf2/HO-1 pathways. Int Immunopharmacol. 2023;114:109619. doi: https://doi.org/10.1016/j.intimp.2022.109619
    | |
  19. Gareri P, Castagna A, Cotroneo AM, Putignano S, De Sarro G, Bruni AC. The role of citicoline in cognitive impairment: pharmacological characteristics, possible advantages, and doubts for an old drug with new perspectives. Clin Interv Aging. 2015;10:1421-9. doi: https://doi.org/10.2147/CIA.S87886
    | |
  20. Cavalu S, Saber S, Ramadan A, Elmorsy EA, Hamad RS, Abdel‐Reheim MA, et al. Unveiling citicoline's mechanisms and clinical relevance in the treatment of neuroinflammatory disorders. FASEB J. 2024;38(17):e70030. doi: https://doi.org/10.1096/fj.202400823R
    | |
  21. Anandan P, Rengarajan S, Venkatachalam S, Pattabi S, Jones S, Prabhu K, et al. Neuroprotection by cerebrolysin and citicoline through the upregulation of Brain-Derived Neurotrophic Factor (BDNF) expression in the affected neural cells: A preliminary clue obtained through an in vitro study. Cureus. 2024;16(2):e54665. doi: https://doi.org/10.7759/cureus.54665
    |
  22. Xu J, Shi Q, Xu W, Zhou Q, Shi R, Ma Y, et al. Metabolic enzyme PDK3 forms a positive feedback loop with transcription factor HSF1 to drive chemoresistance. Theranostics. 2019;9(10):2999-3013. doi: https://doi.org/10.7150/thno.31301
    | |
  23. He L, Lv S, Ma X, Jiang S, Zhou F, Zhang Y, et al. ErbB2 promotes breast cancer metastatic potential via HSF1/LDHA axis-mediated glycolysis. Med Oncol. 2022;39(4):45. doi: https://doi.org/10.1007/s12032-021-01641-4
    | |
  24. Du P, Xu L, Wang Y, Jiao T, Cheng J, Zhang C, et al. Astragaloside IV ameliorates pressure overload-induced heart failure by enhancing angiogenesis through HSF1/VEGF pathway. Heliyon. 2024;10(17):e36687. doi: https://doi.org/10.1016/j.heliyon.2024.e37019
    | |
  25. Fiser O, Muller P. Role of HSF1 in cell division, tumorigenesis and therapy: a literature review. Cell Div. 2025;20(1):1-15. doi: https://doi.org/10.1186/s13008-025-00153-1
    | |
  26. Gerke V, Gavins FNE, Geisow M, Grewal T, Jaiswal JK, Nylandsted J, Rescher U. Annexins-a family of proteins with distinctive tastes for cell signaling and membrane dynamics. Nat Commun. 2024;15(1):1574. doi: https://doi.org/10.1038/s41467-024-45954-0
    | |
  27. Su G, Zhang D, Li T, Pei T, Yang J, Tu S, et al. Annexin A5 derived from matrix vesicles protects against osteoporotic bone loss via mineralization. Bone Res. 2023;11(1):60. doi: https://doi.org/10.1038/s41413-023-00290-9
    | |
  28. Mastropasqua L, Agnifili L, Ferrante C, Sacchi M, Figus M, Rossi G, et al. Citicoline/coenzyme Q10/vitamin B3 fixed combination exerts synergistic protective effects on neuronal cells exposed to oxidative stress. Nutrients. 2022;14(14):2963. doi: https://doi.org/10.3390/nu14142963
    | |
  29. Al-Kuraishy HM, Al-Gareeb AI, Eliwa D, Alexiou A, Papadakis M, Alruwaili M, et al. The mechanistic role of piracetam in the management of vascular dementia. Behav Brain Res. 2025;2025:115551. doi: https://doi.org/10.1016/j.bbr.2025.115551
    | |
  30. de Oliveira AA, Priviero F, Tostes RC, Webb RC, Nunes KP. Dissecting the interaction between HSP70 and vascular contraction: role of Ca2+ handling mechanisms. Sci Rep. 2021;11(1):1420. doi: https://doi.org/10.1038/s41598-021-80966-6
    | |

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2026-06-11

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1.
Danukalo MV. Early neuroprotection: effects on p53 and annexin A5 protein expression in the basal magnocellular nucleus of rats with colchicine-induced injury. Zaporozhye Medical Journal [Internet]. 2026Jun.11 [cited 2026Jun.11];28(3):236-44. Available from: https://zmj.zsmu.edu.ua/article/view/352377

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Vol. 28 No. 3 (2026): Zaporozhye Medical Journal

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Original research

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