THE INFLUENCE OF THE KETONE DIET ON THE LEUKOCYTE FORMULA, STABILITY AND ACTIVITY OF THE GENETIC MATERIAL OF MICE

DOI: 10.22138/2500-0918-2023-20-3-31-41

V.N. Kalaev¹, E.A. Kalaeva¹, I.V. Blazhenov¹,
M.S. Nechaeva², O.Yu. Maltseva³, I.V. Novikova³

¹ Federal State Budgetary Educational Institution of Higher Education
«Voronezh State University», Voronezh, Russian Federation;
² Federal State Budgetary Educational Institution of Higher Education
«N.N. Burdenko Voronezh State Medical University», Voronezh, Russian Federation;
³ Federal State Budget Educational Institution of Higher Education
«Voronezh State University of Engineering Technologies», Voronezh, Russian Federation

Abstract. The effect of a 14-day ketogenic diet on the level of erythrocytes with micronuclei, the activity of nucleolar-forming regions in lymphocytes, and the leukocyte blood count of 8-week-old male C57BL/6 mice was studied. It was found that in animals receiving a ketone diet, the proportion of erythrocytes with Jolly bodies was 18.7 times higher than in mice in the control group receiving a standard diet. The diet did not cause changes in nucleolar activity in lymphocytic cells of the peripheral blood of experimental animals. A slight increase in the level of neutrophils in mice from the experimental group compared with the control was found, probably induced by a change in the biochemical composition of the blood (ketodiet mimics the state after fasting or heavy exercise, which may be accompanied by slight neutrophilia). A significant increase in the proportion of erythrocytes with micronuclei in the peripheral blood of animals of the experimental group may be due to a violation of cell maturation under conditions of severe deficiency of the main energy source for red blood cells — glucose and the inability to use ketone bodies as an energy substrate.

Keywords: ketogenic diet, leukocyte formula, micronuclei, Jolly bodies, erythrocytes, nucleoli

There is no conflict of interest.
Contact details of the corresponding author:
Vladislav N. Kalaev
dr_huixs@mail.ru
Received 03.08.2023
For citation:
Kalaev V.N., Kalaeva E.A., Blazhenov I.V., Nechaeva M.S., Maltseva O.Yu., Novikova I.V. The influence of the ketone diet on the leukocyte formula, stability and activity of the genetic material of mice. [Online] Vestn. Ural. Med. Akad. Nauki. = Journal of Ural Medical Academic Science. 2023, Vol. 20, no. 3, pp. 31–41. DOI: 10.22138/2500-0918-2023-20-3-31-41(In Russ)

REFERENCES
1. Likhachev S.A., Zaytsev I.I., Kulikova S.L., Belaya S.A., Nazarova O.P., Popova M.V. Primenenie ketogennoy diety dlya lecheniya farmakorezistentnoy epilepsii u detey v Respublike Belarus’: pervyy opyt. [The use of the ketogenic diet for the treatment of drug-resistant epilepsy in children in the Republic of Belarus: the first experience]. Nevrologiya i neyrokhirurgiya. Vostochnaya Evropa. 2017; 3: 399–409 (in Russian).
2. Kuchkuntla A.R., Shah M., Velapati S., Gershuni V.M., Rajjo T., Nanda S., Hurt R.T., Mundi M.S. Ketogenic Diet: an Endocrinologist Perspective. Curr Nutr Rep. 2019; 8: 402-410. doi: https://doi.org/10.1007/s13668-019-00297-x.
3. Jensen N.J., Wodschow H.Z., Nilsson M., Rungby J. Effects of Ketone Bodies on Brain Metabolism and Function in Neurodegenerative Diseases. Int J Mol Sci. 2020; 20;21(22): 8767. doi: https://doi.org/10.3390/ijms21228767.
4. Merlotti D., Cosso R., Eller-Vainicher C., Vescini F., Chiodini I., Gennari L., Falchetti A. Energy Metabolism and Ketogenic Diets: What about the Skeletal Health? A Narrative Review and a Prospective Vision for Planning Clinical Trials on this Issue. Int J Mol Sci. 2021; 22 (1): 435. doi: https://doi.org/10.3390/ijms22010435.
5. Valente-Silva P., Lemos C., Köfalvi A., Cunha R.A., Jones J.G. Ketone bodies effectively compete with glucose for neuronal acetyl-CoA generation in rat hippocampal slices. NMR Biomed. 2015; 28: 1111-1116. doi: https://doi.org/10.1002/nbm.3355.
6. Ivannikova E.V., Altashina M.V., Troshina E.A. Ketogennaya dieta: istoriya vozniknoveniya, mekhanizm deystviya, pokazaniya. [The ketogenic diet: history, mechanism of action, indications and contraindications] Problemy endokrinologii. 2021; 68 (1): 49-72. doi:10.14341/probl12724 (in Russian).
7. McCue M.D. Starvation physiology: Reviewing the different strategies animals use to survive a common challenge. Comp. Biochem. Physiol. 2010; 156: 1-18. doi: https://doi.org/10.1016/j.cbpa.2010.01.002.
8. Schwingshackl L., Chaimani A., Hoffmann G., Schwedhelm C., Boeing H. A network meta-analysis on the comparative efficacy of different dietary approaches on glycaemic control in patients with type 2 diabetes mellitus. Eur J Epidemiol. 2018; 33: 157-170. doi: https://doi.org/10.1007/s10654-017-0352-x.
9. Nenartovich I.A. Ketogennaya dieta: biokhimicheskaya osnova i «tekhnika bezopasnosti». [Ketogenic diet: biochemical background and «safety regulations»] Lechebnoe delo: nauchno-prakticheskiy terapevticheskiy zhurnal. 2020; 4 (74): 60-66 (in Russian).
10. Nenartovich I.A. Realii i perspektivy ketogennoy diety v klinicheskoy praktike. [Ketogenic diet realities and prospects in clinical practice] Lechebnoe delo: nauchno-prakticheskiy terapevticheskiy zhurnal. 2021; 1 (76): 66-72 (in Russian).
11. Kaminarskaya Yu.A. Dietoterapiya pri gestatsionnom sakharnom diabete: chto govoryat nauchnye dannye? [Nutritional therapy for gestational diabetes mellitus: what does the evidence say?]. Klinicheskoe pitanie i metabolizm. 2021; 2 (4): 202–214. doi: https://doi.org/10.17816/clinutr105653 (in Russian).
12. Sussman D, Ellegood J, Henkelman M. A gestational ketogenic diet alters maternal metabolic status as well as offspring physiological growth and brain structure in the neonatal mouse. BMC Pregnancy Childbirth. 2013; 13: 198. doi: 10.1186/1471-2393-13-198.
13. Sussman D., van Eede M., Wong M.D., Adamson S.L., Henkelman M. Effects of a ketogenic diet during pregnancy on embryonic growth in the mouse. BMC Pregnancy Childbirth. 2013; 13: 109. doi: 10.1186/1471-2393-13-109.
14. Ilyinsky N.N. Mikroyadernyy analiz v otsenke tsitogeneticheskoy nestabil’nosti [Micronuclear analysis in the assessment of cytogenetic instability] / N.N. Ilyinsky, A.S. Xenz, E.N. Ilyinsky [et al.] Tomsk. Publishing House of TSPU, 2011. 312 p.
15. Kalaev V.N. Mikroyadernyy test bukkal’nogo epiteliya rotovoy polosti cheloveka: monografiya. [Micronucleus test of buccal epithelium of the human oral cavity: monograph] Voronezhskiy gosudarstvennyy universitet. Voronezh: Izdatel’skiy dom VGU; 2016. 136 (in Russian).
16. Howell W. Black D. Controlled silver staining of nucleus organizer regions with a protective colloidal developer: in a one step method. Experientia. 1980; 36: 1014–1015.
17. Theml H., Diem H., Haferlach T. Color Atlas of Hematology. Practical Microscopic and Clinical Diagnosis: 2nd revised edition. Stuttgart. New York: Thieme; 2004. 198.
18. Kalaeva E.A., Artyukhov V.G., Kalaev V.N. Teoreticheskie osnovy i prakticheskoe primenenie matematicheskoy statistiki v biologicheskikh issledovaniyakh i obrazovanii: uchebnik. [Theoretical Foundations and Practical Application of Mathematical Statistics in Biological Research and Education: Textbook] Voronezhskiy gosudarstvennyy universitet. Voronezh: Izdatel’skiy dom VGU; 2016. 284 (in Russian).
19. Kravchenko I.N., Khokhlova O.N., Kravchenko N.N., Puzhalin A.N., D’yachenko I.A., Murashev A.N. Gematologicheskie pokazateli svobodnykh ot patogennoy flory krys CD (Sprague Dawley) i myshey CD1 v norme. [Hematological characters of normal specific pathogen-free cd (Sprague-dawley) rats and cd-1 mice] Biomeditsina. 2008; 2: 20–30 (in Russian).
20. Sears D.A., Udden M.M. Howell–Jolly bodies: a brief historical review. Am. J. Med. Sci. 2011; 343: 407–409.
21. Luzhna L., Kathiria P., Kovalchuk O. Micronuclei in genotoxicity assessment: from genetics to epigenetics and beyond. Front. Genet. 2013; 4: 131–148.
22. Belugin S.N. Ekologicheskaya meditsina: terminologicheskiy slovar’ [Environmental medicine: terminological dictionary] Minsk.: BGMU; 2015. 231 (in Russian).
23. Nuzhnyy V.P., Savchuk S.A. Toksichnost’ alkogol’nykh napitkov i atseton. [Toxicity of alcoholic beverages and acetone] Toksikologicheskiy vestnik. 2005; 5: 22–25 (in Russian).
24. Klenovitskiy P.M., Iolchiev B.S., Zhilinskiy M.A., Bagirov V.A., Onkorova N.T., Grishin V.N. Analiz yadryshek v intaktnykh limfotsitakh perifericheskoy krovi raznykh vidov mlekopitayushchikh.[Analysis of nucleoli in intact peripheral blood lymphocytes in different species of mammals] Dostizheniya nauki i tekhniki APK. 2015; 12: 92-94 (in Russian).
25. Amirov D.R., Tamimdarov B.F., Shageeva A.R. Klinicheskaya gematologiya zhivotnykh: Uchebnoe posobie. [Clinical Animal Hematology: Study Guide] Kazan’.: Tsentr informatsionnykh tekhnologiy KGAVM; 2020. 134 (in Russian).

Authors
Elena A. Kalaeva
Federal State Budgetary Educational Institution of Higher Education «Voronezh State University»
Candidate of Sciences (Biology), Associate Professor, Chair of biophysics and biotechnology
kalaevae@gmail.com
ORCID ID: https://orcid.org/0000-0002-3668-0816
Voronezh, Russian Federation

Vladislav N. Kalaev
Federal State Budgetary Educational Institution of Higher Education «Voronezh State University»
Doctor of Sciences (Biology), Professor, Chair of genetics, cytology and bioengineering
dr_huixs@mail.ru,
ORCID ID: https://orcid.org/0000-0002-4247-4509
Voronezh, Russian Federation

Igor V. Blazhenov
Federal State Budgetary Educational Institution of Higher Education «Voronezh State University»
Student
blazhenov02@mail.ru
ORCID ID: https://orcid.org/0009-0007-8217-3413
Voronezh, Russian Federation

Marina S. Nechaeva
Federal State Budgetary Educational Institution of Higher Education «N.N. Burdenko Voronezh State Medical University» of the Ministry of Healthcare of the Russian Federation
Candidate of Sciences (Biology), Associate Professor, Chair of normal physiology
MAR-Y-ANA@yandex.ru,
ORCID ID: https://orcid.org/0000-0003-4880-6751
Voronezh, Russian Federation

Oksana Yu. Maltseva
Federal State Budget Educational Institution of Higher Education «Voronezh State University of Engineering Technologies»
Candidate of Sciences (Technics) Associate Professor, Chair of biochemistry and biotechnology
ksenia2002@list.ru
ORCID ID: https://orcid.org/0000-0002-3815-123X
Voronezh, Russian Federation

Inna V. Novikova
Federal State Budget Educational Institution of Higher Education «Voronezh State University of Engineering Technologies»
Doctor of Sciences (Technics), Professor, Chair of fermentation and sugar production technologies
noviv@list.ru
ORCID ID: https://orcid.org/0000-0002-2360-5892
Voronezh, Russian Federation