Gender and age-related characteristics of 20S proteasomes response under conditions of calorie restriction

Relevance. The proteasome 20S is a representative of the ubiquitin-protease system, which regulates anabolic and catabolic processes. Searching for ways to control this process is important. Purpose of the study. To study the activation features of the 20S proteasome under low-caloric nutrition conditions and establish relationships between human gender, age and weight characteristics. Materials and methods. The study examined 48 subjects aged 18-75 (11 males and 37 females) received low-energy diet (800-1200 calories/day), fasting therapy, physical therapy and cleansing treatments, as well as physical activity at the Health Center in Maikop, Russia. Blood 20S concentration was measured by enzyme-linked immunosorbent assay, body composition by impedance method. Results. Gender difference in dominant body component determining 20S activity. In females, adipose tissue was dominant; in males, dominant sector not identified. Wellness program led to weight loss and 20S activation regardless of calorie restriction degree. Females under 50 showed activations due to fat while inhibiting muscle and lean masses. Females over 50 had activity in muscle and lean mass while inhibiting fat. Males showed no such patterns in 20S proteasome response. Data suggest different levels of resynthesis, autophagy and apoptosis activation. Calorie restriction is physiological stressor for 20S activation. Conclusion. Hypocaloric diet and fasting with physical therapy can be alternative method for protein pool renewal and resynthesis in humans. Gender and age characteristics should be considered when using these techniques.

proteasome 20S, hypocaloric diet, therapeutic fasting, men, women, body composition

1. Coux O., Zieba B.A., Meiners S. The Proteasome System in Health and Disease. Adv Exp Med Biol. 2020;1233:55-100. doi: 10.1007/978-3-030-38266-7_3.

2. Sahu I., Mali S.M., Sulkshane P., Xu C. et al. The 20S as a stand-alone proteasome in cells can degrade the ubiquitin tag. Nat Commun. 2021;12(1):6173. doi: 10.1038/s41467-021-26427-0.

3. Thomas T., Salcedo-Tacuma D., Smith D.M. Structure, Function, and Allosteric Regulation of the 20S Proteasome by the 11S/PA28 Family of Proteasome Activators. Biomolecules. 2023 Aug 29;13(9):1326. doi: 10.3390/biom13091326.

4. Zhu Y., Shigeyoshi K., Hayakawa Y., Fujiwara S. et al. Acceleration of Protein Degradation by 20S Proteasome-Binding Peptides Generated by In Vitro Artificial Evolution.Int. J. Mol. Sci. 2023;24(24):17486. doi: 10.3390/ijms242417486.

5. Jesenberger V., Jentsch S. Deadly encounter: ubiquitin meets apoptosis. Nat Rev Mol Cell Biol. 2002;3(2):112-21. doi: 10.1038/nrm731.

6. Zhang H.G., Wang J., Yang X., Hsu H.C. et al. Regulation of apoptosis proteins in cancer cells by ubiquitin. Oncogene. 2004;23(11):2009-15. doi: 10.1038/sj.onc.1207373.

7. Yousef A.A., Suliman G.A., Mabrouk M.M. The value of correlation of serum 20S proteasome concentration and percentage of lymphocytic apoptosis in critically ill patients: a prospective observational study. Crit Care. 2010;14(6): R215. doi: 10.1186/cc9340.

8. Renehan A.G., Booth C., Potten C.S. What is apoptosis, and why is it important? BMJ. 2001;322(7301):1536-8. doi: 10.1136/bmj.322.7301.1536.

9. Szondy Z., Sarang Z., Kiss B., Garabuczi É. et al. Anti-inflammatory Mechanisms Triggered by Apoptotic Cells during Their Clearance. Front Immunol. 2017;8:909. doi: 10.3389/fimmu.2017.00909.

10. Martinez M., Reif R., Pappas D. Detection of apoptosis: A review of conventional and novel techniques. Anal Methods. 2010;2: 996-1004. doi: 10.1039/c0ay00247j.

11. Yarovaya G.A., Peshkova E.A., Martynova E.A., Blokhina T.B. The role of proteolytic enzymes in the control of various stages of apoptosis. Laboratory medicine. 2011;11:39-52. (In Russ.)@@ Яровая Г.А., Нешкова Е.А., Мартынова Е.А., Блохина Т.Б. Роль протеолитических ферментов в контроле различных стадий апоптоза. Лабораторная медицина. 2011;11:39-52.

12. D’Arcy M.S. Cell death: a review of the major forms of apoptosis, necrosis and autophagy. Cell Biol Int. 2019;43(6):582-592. doi: 10.1002/cbin.11137.

13. Singh R., Letai A., Sarosiek K. Regulation of apoptosis in health and disease: the balancing act of BCL-2 family proteins. Nat Rev Mol Cell Biol. 2019;20(3):175-193. doi: 10.1038/s41580-018-0089-8.

14. Bhowmick T., Biswas S., Mukherjee A. Cellular response during cellular starvation: a battle forcellular survivability. Cell Biochem Funct. 2024;42: e4101. doi: 10.1002/cbf.4101.

15. Beillevaire D., Migneault F., Turgeon J., Gingras D. et al. Autolysosomes and caspase-3 control the biogenesis and release of immunogenic apoptotic exosomes. Cell Death Dis. 2022;13(2):145. doi: 10.1038/s41419-022-04591-5.

16. Ulas M., Akbas E., Akbas S., Aktemur G. et al. Physiological aspect of apoptosis-regulating microRNAs expressions during fasting. Eur Rev Med Pharmacol Sci. 2023;27(6):2210-2215. doi: 10.26355/eurrev_202303_31754.

17. Yakimovich E.P., Nemtsova V.V., Klyuchnikov D.A. Vozrastnaya anatomiya, fiziologiya i gigiena: uchebnoe posobie dlya vuzov. Vladivostok: Shkola pedagogiki. 2018. 54 p. (In Russ.)@@ Якимович Е.П., Немцова В.В., Ключников Д.А. Возрастная анатомия, физиология и гигиена: учебное пособие для вузов. Владивосток: Школа педагогики. 2018. 54 с.

18. Ahmad O.B., Boschi-Pinto C., Lopez A.D., Murray C.J. et al. Age standardization of rates: a new who standard. World Health Organization. 2001; 31. Available at: https://www.researchgate.net/publication/284696312_Age_Standardization_of_Rates_A_New_WHO_Standard

19. Yu P., Hua Z. To Kill or to Be Killed: How Does the Battle between the UPS and Autophagy Maintain the Intracellular Homeostasis in Eukaryotes? Int J Mol Sci. 2023;24(3):2221. doi: 10.3390/ijms24032221.

20. Li H., Pin S, Zeng Z., Wang M.M. et al. Sex differences in cell death. Ann Neurol. 2005 Aug;58(2):317-21. doi: 10.1002/ana.20538.

21. Lang J.T., McCullough L.D. Pathways to ischemic neuronal cell death: are sex differences relevant? J Transl Med. 2008;6:33. doi: 10.1186/1479-5876-6-33.

22. Abdelbary M., Mohamed R., Gillis E.E., Diaz-Sanders K. et al. Sex differences in apoptosis do not contribute to sex differences in blood pressure or renal T cells in spontaneously hypertensive rats. Front Physiol. 2022;13:1006951. doi: 10.3389/fphys.2022.1006951.

23. Du L., Bayir H., Lai Y., Zhang X. et al. Innate gender-based proclivity in response to cytotoxicity and programmed cell death pathway. J Biol Chem. 2004;279(37):38563-70. doi: 10.1074/jbc.M405461200.

24. Villanueva-Paz M., Cotán D., Garrido-Maraver J., Oropesa-Ávila M. et al. AMPK Regulation of Cell Growth, Apoptosis, Autophagy, and Bioenergetics. Exp Suppl. 2016;107:45-71. doi: 10.1007/978-3-319-43589-3_3.

25. Caro-Maldonado A., Tait S.W., Ramírez-Peinado S., Ricci J.E. et al. Glucose deprivation induces an atypical form of apoptosis mediated by caspase-8 in Bax-, Bak-deficient cells. Cell Death Differ. 2010;17(8):1335-44. doi: 10.1038/cdd.2010.21.

26. Wagner M., Moore D.D. Endoplasmic reticulum stress and glucose homeostasis. Curr Opin Clin Nutr Metab Care. 2011;14(4):367-73. doi: 10.1097/MCO.0b013e32834778d4.

27. Fulda S., Gorman A.M., Hori O., Samali A. Cellular stress responses: cell survival and cell death.Int J Cell Biol. 2010;2010:214074. doi: 10.1155/2010/214074.

28. Petricca S., Flati V., Celenza G., Di Gregorio J. et al. Tebuconazole and Econazole Act Synergistically in Mediating Mitochondrial Stress, Energy Imbalance, and Sequential Activation of Autophagy and Apoptosis in Mouse Sertoli TM4 Cells: Possible Role of AMPK/ULK1 Axis. Toxicol Sci. 2019 May 1;169(1):209-223. doi: 10.1093/toxsci/kfz031.

29. Nüchel J., Tauber M., Nolte J.L., Mörgelin M. et al. An mTORC1-GRASP55 signaling axis controls unconventional secretion to reshape the extracellular proteome upon stress. Mol Cell. 2021;81(16):3275-3293.e12. doi: 10.1016/j.molcel.2021.06.017.

30. Uriarte M., Sen Nkwe N., Tremblay R., Ahmed O. et al. Starvation-induced proteasome assemblies in the nucleus link amino acid supply to apoptosis. Nat Commun. 2021;12(1):6984. doi: 10.1038/s41467-021-27306-4.

31. Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol. 2007;35(4):495-516. doi: 10.1080/01926230701320337.

32. Qi Z., Chen L. Endoplasmic Reticulum Stress and Autophagy. Adv Exp Med Biol. 2019;1206:167-177. doi: 10.1007/978-981-15-0602-4_8.

33. Mei Y., Glover K., Su M., Sinha S.C. Conformational flexibility of BECN1: Essential to its key role in autophagy and beyond. Protein Sci. 2016;25(10):1767-85. doi: 10.1002/pro.2984.

34. Tkhakushinov I.A., Lysenkov S.Р., Korchazhkina N.B., Muzhenya D.V. et al. Gender and age peculiarities of autophagy in men and women induced by a comprehensive wellness program: a prospective observational study. Kuban Scientific Medical Bulletin. 2024;31(2):95-106. (In Russ.) doi: 10.25207/1608-6228-2024-31-2-95-106.@@ Тхакушинов И.А., Лысенков С.П., Корчажкина Н.Б., Муженя Д.В. с соавт. Гендерные и возрастные особенности аутофагии у мужчин и женщин в условиях проведения комплексной оздоровительной программы: проспективное наблюдательное исследование. Кубанский научный медицинский вестник. 2024;31(2):95-106. doi: 10.25207/1608-6228-2024-31-2-95-106.

35. Ferhat M., Funai K., Boudina S. Autophagy in Adipose Tissue Physiology and Pathophysiology. Antioxid Redox Signal. 2019;31(6):487-501. doi: 10.1089/ars.2018.7626.

36. Kraus W.E., Bhapkar M., Huffman K.M., Pieper C.F. et al. CALERIE Investigators. 2 years of calorie restriction and cardiometabolic risk (CALERIE): exploratory outcomes of a multicentre, phase 2, randomised controlled trial. Lancet Diabetes Endocrinol. 2019;7(9):673-683. doi: 10.1016/S2213-8587(19)30151-2.