Gender and age peculiarities of autophagy in men and women induced by a comprehensive wellness program: a prospective observational study

Background. Autophagy refers to an evolutionary physiological mechanism of cell self-purification. The use of its positive effects in regenerative and preventive medicine implies this process to be investigated under the conditions of various therapeutic factors.
Objective. To investigate the influence of gender, age and somatic-morphological factors on the autophagy processes induced by a comprehensive wellness program as expressed by the marker protein Beclin-1.
Methods. A prospective observational study involved 107 patients to be monitored before and after comprehensive wellness procedures. All study participants underwent a wellness program in the clinic of OOO Centr Zdorovie (Maykop, Russia). The study was conducted in the periods of September‒December 2020 and January 2021. The study involved measuring the concentration of Beclin-1 protein in the blood before and 12 days after the wellness procedures. In order to characterize the degree of activation of the autophagy process, the index “delta-Beclin-1” was introduced (difference between the initial level of Beclin-1 protein concentration and the level registered after 12 days). The autophagy response was evaluated in terms of its upward or downward direction (positive/negative delta- Beclin-1 value, respectively). The effect of the comprehensive wellness procedures on the functional systems of the organism was determined in accordance with the laboratory tests (complete blood count, lipid profile), morphometric evaluation, and the assessment of body composition by impedance monitoring. The obtained data were processed using SPSS Statistics 26.0 (IBM, USA).
Results. Monitoring of Beclin-1 protein concentration revealed that basic autophagy activity is age-specific and significantly lower in individuals over 60 years as compared to both individuals under 60 years and healthy persons. This correlation with age appears more pronounced in obese individuals (r = -0.59). The basic activity of autophagy in men was 30% lower than in women, however, after a set of wellness procedures the mentioned response in men increased by 7%. The study found that not all patients responded unidirectionally to the comprehensive wellness procedures: 49 out of 77 patients showed an increase in Beclin-1 protein concentration, while 28 — a decrease. Patients who participated in the wellness program demonstrated negative correlation between the level of Beclin-1 protein expression and age, while those responding with an increase in Beclin-1 protein level — with body mass index, weight and fat mass. The source of autophagy activity in men under 60 years can refer to fat mass, and in women — lean body mass, especially in the mature age of period II (36‒60 years).
Conclusion. The factors of basic autophagy and autophagy induced by the complex of wellness procedures include age, gender and body weight. These factors obtain different significance at different age periods. A comprehensive wellness program can serve as an alternative to existing pharmacological methods for activating autophagy in humans.

1. Ohsumi Y. Historical landmarks of autophagy research. Cell Res. 2014;24(1):9–23. https://doi.org/10.1038/cr.2013.169

2. Corti O, Blomgren K, Poletti A, Beart PM. Autophagy in neurodegeneration: New insights underpinning therapy for neurological diseases. J Neurochem. 2020;154(4):354–371. https://doi.org/10.1111/jnc.15002

3. Fleming A, Bourdenx M, Fujimaki M, Karabiyik C, Krause GJ, Lopez A, Martín-Segura A, Puri C, Scrivo A, Skidmore J, Son SM, Stamatakou E, Wrobel L, Zhu Y, Cuervo AM, Rubinsztein DC. The different autophagy degradation pathways and neurodegeneration. Neuron. 2022;110(6):935–966. https://doi.org/10.1016/j.neuron.2022.01.017

4. Xu HD, Qin ZH. Beclin 1, Bcl-2 and Autophagy. AdvExp Med Biol. 2019;1206:109–126. https://doi.org/10.1007/978-981-15-0602-4_5

5. Kong Z, Yao T. Role for autophagy-related markers Beclin-1 and LC3 in endometriosis. BMC Womens Health. 2022;22(1):264. https://doi.org/10.1186/s12905-022-01850-7

6. Bhardwaj JK, Paliwal A, Saraf P, Sachdeva SN. Role of autophagy in follicular development and maintenance of primordial follicular pool in the ovary. J Cell Physiol. 2022;237(2):1157–1170. https://doi.org/10.1002/jcp.30613

7. Lin F, Zhu YT, Qin ZH. Biomarkers of Autophagy. AdvExp Med Biol. 2021;1208:265–287. https://doi.org/10.1007/978-981-16-2830-6_12

8. Mei Y, Su M, Sanishvili R, Chakravarthy S, Colbert CL, Sinha SC. Identification of BECN1 and ATG14 Coiled-Coil Interface Residues That Are Important for Starvation-Induced Autophagy. Biochemistry. 2016;55(30):4239–4253. https://doi.org/10.1021/acs.biochem.6b00246

9. Nechushtai L, Frenkel D, Pinkas-Kramarski R. Autophagy in Parkinson’s Disease. Biomolecules. 2023;13(10):1435. https://doi.org/10.3390/biom13101435

10. Kanno H, Handa K, Murakami T, Aizawa T, Ozawa H. Chaperone-Mediated Autophagy in Neurodegenerative Diseases and Acute Neurological Insults in the Central Nervous System. Cells. 2022;11(7):1205. https://doi.org/10.3390/cells11071205

11. Marzoog BA. Autophagy as an Anti-senescent in Aging Neurocytes. Curr Mol Med. 2024;24(2):182–190. https://doi.org/10.2174/1566524023666230120102718

12. Marzoog BA. Endothelial Cell Aging and Autophagy Dysregulation. Cardiovasc Hematol Agents Med Chem. 2024;22. https://doi.org/10.2174/0118715257275690231129101408

13. Guo J, Chiang WC. Mitophagy in aging and longevity. IUBMB Life. 2022;74(4):296–316. https://doi.org/10.1002/iub.2585

14. Kim YA, Kim YS, Oh SL, Kim HJ, Song W. Autophagic response to exercise training in skeletal muscle with age. J PhysiolBiochem. 2013;69(4):697–705. https://doi.org/10.1007/s13105-013-0246-7

15. Xie G, Jin H, Mikhail H, Pavel V, Yang G, Ji B, Lu B, Li Y. Autophagy in sarcopenia: Possible mechanisms and novel therapies. Biomed Pharmacother. 2023;165:115147. https://doi.org/10.1016/j.biopha.2023.115147

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

17. Kraus WE, Bhapkar M, Huffman KM, Pieper CF, Krupa Das S, Redman LM, Villareal DT, Rochon J, Roberts SB, Ravussin E, Holloszy JO, Fontana L; 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. https://doi.org/10.1016/S2213-8587(19)30151-2

18. Teong XT, Liu K, Vincent AD, Bensalem J, Liu B, Hattersley KJ, Zhao L, Feinle-Bisset C, Sargeant TJ, Wittert GA, Hutchison AT, Heilbronn LK. Intermittent fasting plus early time-restricted eating versus calorie restriction and standard care in adults at risk of type 2 diabetes: a randomized controlled trial. Nat Med. 2023;29(4):963–972. https://doi.org/10.1038/s41591-023-02287-7

19. Lu DL, Ma Q, Wang J, Li LY, Han SL, Limbu SM, Li DL, Chen LQ, Zhang ML, Du ZY. Fasting enhances cold resistance in fish through stimulating lipid catabolism and autophagy. J Physiol. 2019;597(6):1585–1603. https://doi.org/10.1113/JP277091

20. Shabkhizan R, Haiaty S, Moslehian MS, Bazmani A, Sadeghsoltani F, SaghaeiBagheri H, Rahbarghazi R, Sakhinia E. The Beneficial and Adverse Effects of Autophagic Response to Caloric Restriction and Fasting. AdvNutr. 2023;14(5):1211–1225. https://doi.org/10.1016/j.advnut.2023.07.006

21. Sadeghi A, Niknam M, Momeni-Moghaddam MA, Shabani M, Aria H, BastinA, Teimouri M, Meshkani R, Akbari H. Crosstalk between autophagy and insulin resistance: evidence from different tissues. EurJMedRes. 2023;28(1):456. https://doi.org/10.1186/s40001-023-01424-9

22. Залевская М.А. Возрастная периодизация в Российской Федерации в современных условиях. Международный научно-исследовательский журнал. 2020;13-3(102):17–20. https://doi.org/10.23670/IRJ.2020.102.12.073

23. Zalevskaya MA. Contemporary conditions of age groups in the Russian Federation. Mezhdunarodnyy Nauchno-Issledovatel’skiy Zhurnal. 2020;13-3(102):17–20 (In Russ). https://doi.org/10.23670/IRJ.2020.102.12.073

24. Nghia HL, Van Tiep N. About the Operating Principles of System DDFAO “(Dépistage et DiagnostiqueFonctionnelAssisté Par Ordinateur)”. The Third International Conference on the Development of Biomedical Engineering in Vietnam. 2010;94–97. https://doi.org/10.1007/978-3-642-12020-6_23

25. Luévano-Martínez LA, Forni MF, Peloggia J, Watanabe IS, Kowaltowski AJ. Calorie restriction promotes cardiolipin biosynthesis and distribution between mitochondrial membranes. Mech Ageing Dev. 2017;162:9–17. https://doi.org/10.1016/j.mad.2017.02.004

26. Barbosa MC, Grosso RA, Fader CM. Hallmarks of Aging: An Autophagic Perspective. Front Endocrinol (Lausanne). 2019;9:790. https:// doi.org/10.3389/fendo.2018.00790

27. Ding S, Jiang J, Zhang G, Bu Y, Zhang G, Zhao X. Resveratrol and caloric restriction prevent hepatic steatosis by regulating SIRT1-autophagy pathway and alleviating endoplasmic reticulum stress in highfat diet-fed rats. PLoS One. 2017;12(8):e0183541. https://doi.org/10.1371/journal.pone.0183541

28. Yao ZQ, Zhang X, Zhen Y, He XY, Zhao S, Li XF, Yang B, Gao F, Guo FY, Fu L, Liu XZ, Duan CZ. A novel small-molecule activator of Sirtuin-1 induces autophagic cell death/mitophagy as a potential therapeutic strategy in glioblastoma. Cell Death Dis. 2018;9(7):767. https://doi.org/10.1038/s41419-018-0799-z

29. Lee BB, Kim Y, Kim D, Cho EY, Han J, Kim HK, Shim YM, Kim DH. Metformin and tenovin-6 synergistically induces apoptosis through LKB1-independent SIRT1 down-regulation in non-small cell lung cancer cells. J Cell Mol Med. 2019;23(4):2872–2889. https://doi.org/10.1111/jcmm.14194

30. Aventaggiato M, Barreca F, Vitiello L, Vespa S, Valente S, Rotili D, Mai A, Lotti LV, Sansone L, Russo MA, Bizzarri M, Ferretti E, Tafani M. Role of SIRT3 in Microgravity Response: A New Player in Muscle Tissue Recovery. Cells. 2023;12(5):691. https://doi.org/10.3390/cells12050691

31. Yin S, Liu L, Gan W. The Roles of Post-Translational Modifications on mTOR Signaling. Int J Mol Sci. 2021;22(4):1784. https://doi.org/10.3390/ijms22041784

32. Ganley IG, Lam du H, Wang J, Ding X, Chen S, Jiang X. ULK1.ATG13. FIP200 complex mediates mTOR signaling and is essential for autophagy. J Biol Chem. 2009;284(18):12297–12305. https://doi.org/10.1074/jbc.M900573200

33. Hosokawa N, Hara T, Kaizuka T, Kishi C, Takamura A, Miura Y, Iemura S, Natsume T, Takehana K, Yamada N, Guan JL, Oshiro N, Mizushima N. Nutrient-dependent mTORC1 association with the ULK1-Atg13-FIP200 complex required for autophagy. MolBiol Cell. 2009;20(7):1981–1991. https://doi.org/10.1091/mbc.e08-12-1248

34. Das JK, Banskota N, Candia J, Griswold ME, Orenduff M, de Cabo R, Corcoran DL, Das SK, De S, Huffman KM, Kraus VB, Kraus WE, Martin CK, Racette SB, Redman LM, Schilling B, Belsky DW, Ferrucci L. Calorie restriction modulates the transcription of genes related to stress response and longevity in human muscle: The CALERIE study. AgingCell. 2023;22(12):e13963. https://doi.org/10.1111/acel.13963