The specificity of iNOS expression indicators in the basal magnocellular nucleus of rats under early pathogenetic correction in experimental neurodestruction

Authors

DOI:

https://doi.org/10.14739/2310-1210.2024.5.309732

Keywords:

neuroprotection, nitric oxide synthase, nitrosative stress, neurodegeneration, brain, neurons, cognitive function disorders

Abstract

Aim. To characterize iNOS expression indicators in the basal magnocellular nucleus of rats during early pathogenetic correction of neurodegeneration induced by intracerebroventricular colchicine administration.

Materials and methods. The study was conducted using 50 male Wistar rats aged 10–11 months, which were divided into 5 experimental groups (n = 10). The control animals (group 1) were administered a 0.9 % NaCl solution intracerebroventricularly, while the other experimental group rats (groups 2–5) received a colchicine solution in the same manner. The following day, the animals from groups 3–5 were initiated early pathogenetic correction with citicoline (group 3), thiocetam (group 4), and HSF-1 (group 5) lasting 14 days. All the experimental animals (groups 1–5) were then euthanized with sodium thiopental, and their brains were extracted for histochemical, immunofluorescent, and biochemical examinations.

Results. The study has demonstrated that intracerebroventricular administration of colchicine to rats was accompanied by morphological signs of neurodegeneration in the basal magnocellular nucleus and characterized by a significantly smaller area of chromaffin substance in the neurons of this structure by 39 % as compared to the control animals. At the same time, early pathogenetic correction of colchicine-induced neurodegeneration was associated with significantly larger values of Nissl substance area of the basal magnocellular nucleus neurons compared to the corresponding values in animals that did not receive the correction. Additionally, the nitrite level in the brain homogenates of rats administered colchicine without correction (group 2) exceeded the control (group 1) by almost 7 times, while the indicators in experimental groups 3, 4, and 5 exceeded it by 3.5, 2.9, and 3.8 times, respectively. However, no statistical differences were found between the control group and the correction groups in terms of nitrite content. Evaluating the expression of iNOS (the area of immunopositive cells and corrected total cell fluorescence, CTCF) in the basal magnocellular nucleus of the experimental rats it has been shown that the area was most affected in group 5 (HSF-1 correction), exceeding the control parameter by 18.9 %, group 3 (citicoline correction) by 14.7 %, and group 4 (thiocetam correction) by 17.1 %, with no statistical differences compared to group 2 (colchicine administration without correction). Meanwhile, the CTCF of iNOS+-cells in the basal magnocellular nucleus of the experimental animals was the highest in group 2 significantly exceeding the corresponding parameters in the control and correction groups. No significant differences were found between the control and correction groups in this parameter. Additionally, it is noteworthy that intracerebroventricular administration of colchicine to rats, compared to control animals, was associated with a significant double the number of iNOS+ cells in the basal magnocellular nucleus. However, early pathogenetic correction in groups 3-5 did not significantly affect the number of iNOS+ cells in the studied structure, as this parameter did not statistically differ from group 2, although significantly exceeding the corresponding parameter in the control group (group 1).

Conclusions. Early pathogenetically substantiated correction with citicoline, thiocetam, and HSF-1 in colchicine-induced neurodegeneration in the basal magnocellular nucleus of experimental rats is accompanied by an increase in the area of chromaffin substance compared to rats that received intracerebroventricular colchicine without correction, as well as a reduction in nitrite levels in brain homogenates to the control level (sham-operated animals). In the basal magnocellular nucleus of experimental rats, under the influence of early pathogenetically substantiated correction of experimental neurodegeneration, iNOS expression indicators (area of immunopositive cells and CTCF) vary depending on the neuroprotectant used. The number of iNOS+ cells in the basal magnocellular nucleus of experimental rats in the correction groups does not change compared to the group that received intracerebroventricular colchicine without correction and is statistically higher than the corresponding indicator in the control rats.

Author Biographies

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

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

Yu. M. Kolesnyk, Zaporizhzhia State Medical and Pharmaceutical University, Ukraine

MD, PhD, DSc, Professor of the Department of Pathological Physiology with the Course of Normal Physiology, Rector of Zaporizhzhia State Medical and Pharmaceutical University, Honored Science and Technology Figure of Ukraine

References

Oswald MJ, Han Y, Li H, Marashli S, Oglo DN, Ojha B, et al. Cholinergic basal forebrain nucleus of Meynert regulates chronic pain-like behavior via modulation of the prelimbic cortex. Nature Communications. 2022;13(1):5014. https://doi.org/10.1038/s41467-022-32558-9

Tiepolt S, Meyer PM, Patt M, Deuther-Conrad W, Hesse S, Barthel H, et al. PET Imaging of Cholinergic Neurotransmission in Neurodegenerative Disorders. J Nucl Med. 2022;63(Suppl 1):33S-44S. doi: https://doi.org/10.2967/jnumed.121.263198

Aisen PS, Jimenez-Maggiora GA, Rafii MS, Walter S, Raman R. Early-stage Alzheimer disease: getting trial-ready. Nat Rev Neurol. 2022;18(7):389-99. doi: https://doi.org/10.1038/s41582-022-00645-6

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

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.

Sleigh JN, Rossor AM, Fellows AD, Tosolini AP, Schiavo G. Axonal transport and neurological disease. Nat Rev Neurol. 2019;15(12):691-703. doi: https://doi.org/10.1038/s41582-019-0257-2

Guo W, Stoklund Dittlau K, Van Den Bosch L. Axonal transport defects and neurodegeneration: Molecular mechanisms and therapeutic implications. Semin Cell Dev Biol. 2020;99:133-50. doi: https://doi.org/10.1016/j.semcdb.2019.07.010

Brown A. Slow axonal transport: stop and go traffic in the axon. Nat Rev Mol Cell Biol. 2000;1(2):153-6. doi: https://doi.org/10.1038/35040102

Balez R, Ooi L. Getting to NO Alzheimer's Disease: Neuroprotection versus Neurotoxicity Mediated by Nitric Oxide. Oxid Med Cell Longev. 2016;2016:3806157. doi: https://doi.org/10.1155/2016/3806157

Jasielski P, Piędel F, Piwek M, Rocka A, Petit V, Rejdak K. Application of Citicoline in Neurological Disorders: A Systematic Review. Nutrients. 2020;12(10):3113. doi: https://doi.org/10.3390/nu12103113

Adibhatla RM, Hatcher JF. Citicoline decreases phospholipase A2 stimulation and hydroxyl radical generation in transient cerebral ischemia. J Neurosci Res. 2003;73(3):308-15. doi: https://doi.org/10.1002/jnr.10672

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.

Kim JY, Barua S, Huang MY, Park J, Yenari MA, Lee JE. Heat Shock Protein 70 (HSP70) Induction: Chaperonotherapy for Neuroprotection after Brain Injury. Cells. 2020;9(9):2020. doi: https://doi.org/10.3390/cells9092020

Paxinos G, Watson C. The Rat Brain in stereotaxic coordinates. London: Elsevier, Academic Press; 2018.

Zhu Y, Liu F, Zou X, Torbey M. Comparison of unbiased estimation of neuronal number in the rat hippocampus with different staining methods. J Neurosci Methods. 2015;254:73-9. doi: https://doi.org/10.1016/j.jneumeth.2015.07.022

Santa Cruz Biotechnology. Immunofluorescence Cell Staining [Internet]. www.scbt.com. Available from: https://www.scbt.com/resources/protocols/immunofluorescence-cell-staining

Miranda KM, Espey MG, Wink DA. A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide. 2001;5(1):62-71. doi: https://doi.org/10.1006/niox.2000.0319

Holovanova IA, Bielikova IV, Liakhova NO. Osnovy medychnoi statystyky [Basics of medical statistics]. Poltava: PDMU; 2017. Ukrainian. Available from: https://repository.pdmu.edu.ua/handle/123456789/10614

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

Kumar A, Dogra S. Neuroprotective effect of carvedilol, an adrenergic antagonist against colchicine induced cognitive impairment and oxidative damage in rat. Pharmacol Biochem Behav. 2009 Mar;92(1):25-31. doi: https://doi.org/10.1016/j.pbb.2008.10.005

Hurtado O, Hernández-Jiménez M, Zarruk JG, Cuartero MI, Ballesteros I, Camarero G, et al. Citicoline (CDP-choline) increases Sirtuin1 expression concomitant to neuroprotection in experimental stroke. J Neurochem. 2013;126(6):819-26. doi: https://doi.org/10.1111/jnc.12269

Synoradzki K, Grieb P. Citicoline: A Superior Form of Choline? Nutrients. 2019;11(7):1569. doi: https://doi.org/10.3390/nu11071569

Hurtado O, Moro MA, Cárdenas A, Sánchez V, Fernández-Tomé P, Leza JC, et al. Neuroprotection afforded by prior citicoline administration in experimental brain ischemia: effects on glutamate transport. Neurobiol Dis. 2005;18(2):336-45. doi: https://doi.org/10.1016/j.nbd.2004.10.006

Qureshi I, Endres JR. Citicoline: a novel therapeutic agent with neuroprotective, neuromodulatory, and neuroregenerative properties. Natural Medicine J. 2010;2(6):11-25.

Belenichev IF, Cherniy V, Nahorna E, Pavlov S, Cherniy T, Bukhtiyarova N, et al. Nejroprotekcija i nejroplastichnost [Neuroprotection and neuroplasticity]. Kyiv: Logos; 2015. Russian.

Belenichev IF, Gorbacheva SV, Bukhtiyarova NV, Levich SV. Dynamics of changes in the concentration of heat shock protein (HSP70) in the cerebral cortex and hippocampus in experimental violation of cerebral circulation: the ability to regulate this process through positive modulation of thiol-disulfide system. Biological Markers and Guided Therapy. 2016;3(1):107-14. doi: http://dx.doi.org/10.12988/bmgt.2016.6311

Pourbagher-Shahri AM, Farkhondeh T, Talebi M, Kopustinskiene DM, Samarghandian S, Bernatoniene J. An overview of no signaling pathways in aging. Molecules. 2021;26(15):4533. doi: https://doi.org/10.3390/molecules26154533

Doherty GH. Nitric oxide in neurodegeneration: potential benefits of non-steroidal anti-inflammatories. Neurosci Bull. 2011;27(6):366-82. doi:

Additional Files

Published

2024-10-04

How to Cite

1.
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 [Internet]. 2024Oct.4 [cited 2024Nov.15];26(5):379-86. Available from: http://zmj.zsmu.edu.ua/article/view/309732