Chronic stress-induced apoptosis is mitigated by young mitochondria transplantation in the prefrontal cortex of aged rats

Document Type : Original Article


1 Drug Applied Research, Tabriz University of Medical Sciences, Tabriz, Iran

2 Neurosciences Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

3 Department of Biomedical Education, California Health Sciences University, College of Osteopathic Medicine, Clovis, CA, USA


Objective(s): Apoptosis is common and often comorbid with aging and stress-related mood disorders. Evidence suggests that fresh mitochondria could reverse age-related dysfunctions in organs, especially in the brain. Therefore, this study investigated the effect of young mitochondria administration on the apoptosis process in the prefrontal cortex (PFC) of aged rats exposed to chronic stress. 
Materials and Methods: Aged (22 months old) male rats were randomly assigned into four groups: aged control (AC), aged rats treated with young mitochondria (A+M), aged rats subjected to chronic stress for four weeks (A+St), and aged rats subjected to chronic stress and treated with young mitochondria (A+St+M). A+M and A+St+M groups received a single ICV injection (10 μl) of fresh mitochondria isolated from the brain of young rats for five minutes (2 µl/min). Finally, the levels of Malondialdehyde (MDA), Cytochrome c (Cyt c), Bax, Bcl-2, and Caspase-3 expression were investigated in the PFC.
Results: Young mitochondria administration reduced neuronal apoptosis in the PFC, associated with down-regulation of MDA, Bax, and Caspase-3 and up-regulation of Bcl-2. Moreover, fresh mitochondria partially improved the chronic stress-induced mitochondrial dysfunction in aged rats, as indicated by reduced cytochrome c (Cyt c) release from the mitochondria.
Conclusion: These results suggest mitotherapy could reverse cell viability and mitochondrial dysfunction-induced apoptosis in the PFC tissue of aged rats subjected to stressful stimuli.


Main Subjects

1. Yamaguchi N, Nakajima N, Okada S, Yuri K. Effects of aging on stress-related responses of serotonergic neurons in the dorsal raphe nucleus of male rats. Neurobiol Stress 2016; 3: 43–51. 
2. McEwen BS and Morrison JH. The brain on stress: Vulnerability and plasticity of the prefrontal cortex over the life course. Neuron 2013; 79: 16–29. 
3. Ghaffari-Nasab A, Badalzadeh R, Mohaddes G, Alipour MR. Young plasma administration mitigates depression-like behaviours in chronic mild stress-exposed aged rats by attenuating apoptosis in prefrontal cortex. Exp Physiol 2021; 106: 1621–1630. 
4. Srivastava S. Emerging therapeutic roles for NAD+ metabolism in mitochondrial and age-related disorders. Clin Transl Med 2016 51 2016; 5: 1–11. 
5. Wallace DC, Fan W, Procaccio V. Mitochondrial energetics and therapeutics. Annu Rev Pathol 2010; 5:297-348. 
6. Sun N, Youle RJ, Finkel T. The mitochondrial basis of aging. Mol Cell 2016; 61: 654–666. 
7. Srivastava S. The mitochondrial basis of aging and age-related disorders. Genes 2017; 8. 
8. Haas RH. Mitochondrial dysfunction in aging and diseases of aging. Biology 2019; 8: 48. 
9. Guo CY, Sun L, Chen XP, Zhang DS. Oxidative stress, mitochondrial damage and neurodegenerative diseases. Neural Regen Res 2013; 8: 2003-2014. 
10. Allen J, Romay-Tallon R, Brymer KJ, Caruncho HJ, Kalynchuk LE. Mitochondria and mood: Mitochondrial dysfunction as a key player in the manifestation of depression. Front Neurosci 2018; 12: 1-13.
11. Bajpai A, Verma AK, Srivastava M, Srivastava R. Oxidative stress and major depression. J Clin Diagn Res 2014; 8: CC04–CC07. 
12. Zhao Z, Yu Z, Hou Y, Zhang L, Fu A. Improvement of cognitive and motor performance with mitotherapy in aged mice. Int J Biol Sci 2020; 16: 849–858. 
13. Nascimento-Dos-santos G, De-Souza-ferreira E, Linden R, Galina A, Petrs-Silva H. Mitotherapy: Unraveling a promising treatment for disorders of the central nervous system and other systemic conditions. Cells 2021; 10: 1827. 
14. Chi H, Chang H-Y, Sang T-K. Neuronal cell death mechanisms in major neurodegenerative Diseases. Int J Mol Sci 2018; 19: 1-18. 
15. Dygalo NN, Kalinina TS, Bulygina V V, Shishkina GT. Increased expression of the anti-apoptotic protein bcl-xl in the brain is associated with resilience to stress-induced depression-like behavior. Cell Mol Neurobiol 2012; 32: 767–776. 
16. Zhao Z, Zhang L, Guo XD, Cao LL, Xue TF, Zhao XJ, et al. Rosiglitazone exerts an anti-depressive effect in unpredictable chronic mild-stress-induced depressive mice by maintaining essential neuron autophagy and inhibiting excessive astrocytic apoptosis. Front Mol Neurosci 2017; 10: 1-16.
17. Eskes R, Antonsson B, Osen-Sand A, Montessuit S, Richter C, Sadoul R, et al. Bax-induced cytochrome c release from mitochondria is independent of the permeability transition pore but highly dependent on Mg2+ Ions. J Cell Biol 1998; 143: 217-224. 
18. Hongmei Z. Extrinsic and intrinsic apoptosis signal pathway review. In: Apoptosis and medicine. InTechOpen 2012. 
19. Reutzel M, Grewal R, Dilberger B, Silaidos C, Joppe A, and Eckert GP. Cerebral mitochondrial function and cognitive performance during aging: A longitudinal study in NMRI mice. Oxid Med Cell Longev 2020; 2020:1–12. 
20. Ghaffari-Nasab A, Badalzadeh R, Mohaddes G, Javani G, Ebrahimi-kalan A, Alipour MR. Young plasma induces antidepressant-like effects in aged rats subjected to chronic mild stress by suppressing indoleamine 2,3-dioxygenase enzyme and kynurenine pathway in the prefrontal cortex. Neurochem Res 2022; 47: 358–371. 
21. Javani G, Babri S, Farajdokht F, Ghaffari-Nasab A, Mohaddes G. Mitochondrial transplantation improves anxiety- and depression-like behaviors in aged stress-exposed rats. Mech Ageing Dev 2022; 202: 111632. 
22. Spijker S. Dissection of Rodent Brain Regions. In: Neuromethods. Springer; 2011; 13–26. 
23. Paxinos G and Franklin KBJ. The mouse brain in stereotaxic coordinates: Hard cover edition. Vol. 2nd Editio, Academic Press. Elsevier; 2001. 
24. Liu J, Wang X, Mori A. Immobilization stress-induced anti-oxidant defense changes in rat plasma: effect of treatment with reduced glutathione. Int J Biochem 1994; 26: 511–517. 
25. Maes M, Galecki P, Chang YS, Berk M. A review on the oxidative and nitrosative stress (O&NS) pathways in major depression and their possible contribution to the (neuro)degenerative processes in that illness. Prog Neuropsychopharmacol Biol Psychiatry 2011; 35: 676–692. 
26. Kregel KC and Zhang HJ. An integrated view of oxidative stress in aging: Basic mechanisms, functional effects, and pathological considerations. Am J Physiol - Regul Integr Comp Physiol 2007; 292: 18–36. 
27. Bouayed J, Rammal H, Soulimani R. Oxidative stress and anxiety Relationship and cellular pathways. Oxid Med Cell Longev 2009; 2: 63–67. 
28. Bouayed J. Relationship between oxidative stress and anxiety: Emerging role of anti-oxidants within therapeutic or preventive approaches. In: anxiety disorders. InTech 2011; 2: 27-38. 
29. Bouayed J. Polyphenols: A potential new strategy for the prevention and treatment of anxiety and depression. Curr Nutr Food Sci 2010; 6: 13–18. 
30. Liguori I, Russo G, Curcio F, Bulli G, Aran L, Della-Morte D, et al. Oxidative stress, aging, and diseases. Clin Interv Aging 2018; 13: 757. 
31. Michel TM, Frangou S, Thiemeyer D, Camara S, Jecel J, Nara K, et al. Evidence for oxidative stress in the frontal cortex in patients with recurrent depressive disorder--a postmortem study. Psychiatry Res 2007; 151: 145–150. 
32. Lee JM, Hwang JW, Kim MJ, Jung SY, Kim KS, Ahn EH, et al. Mitochondrial transplantation modulates inflammation and apoptosis, alleviating tendinopathy both in vivo and in vitro. Anti-oxidants 2021; 10: 1-22. 
33. Shi C, Guo H, Liu X. Platelet mitochondria transplantation rescues hypoxia/reoxygenation-induced mitochondrial dysfunction and neuronal cell death involving the FUNDC2/PIP3/Akt/FOXO3a axis. Cell Transplant 2021; 30: 1–9. 
34. Aharoni-Simon M, Ben-Yaakov K, Sharvit-Bader M, Raz D, Haim Y, Ghannam W, et al. Oxidative stress facilitates exogenous mitochondria internalization and survival in retinal ganglion precursor-like cells. Sci Rep 2022; 12: 5122-5130. 
35. Lin M-W, Fang S-Y, Hsu J-YC, Huang C-Y, Lee P-H, Huang C-C, et al. Mitochondrial transplantation attenuates neural damage and improves locomotor function after traumatic spinal cord injury in rats. Front Neurosci 2022; 16:800883
36. Tower J. Programmed cell death in aging. Ageing Res Rev 2015; 23: 90-100. 
37. Kwak H-B. Effects of aging and exercise training on apoptosis in the heart. J Exerc Rehabil 2013; 9: 212-219. 
38. Wang X, Bonventre J, Parrish A. The aging kidney: Increased susceptibility to nephrotoxicity. Int J Mol Sci 2014; 15:15358–15376. 
39. Jeong SY and Seol DW. The role of mitochondria in apoptosis. BMB Rep 2008; 41: 11–22. 
40. Abu-Qare AW and Abou-Donia MB. Biomarkers of apoptosis: Release of cytochrome c, activation of caspase-3, induction of 8-hydroxy-2’-deoxyguanosine, increased 3-nitrotyrosine, and alteration of p53 gene. J Toxicol Environ Health B Crit Rev 2001; 4: 313–332. 
41. Nechushtan A, Smith CL, Lamensdorf I, Yoon SH, Youle RJ. Bax and Bak coalesce into novel mitochondria-associated clusters during apoptosis. J Cell Biol 2001; 153: 1265–1276. 
42. Acehan D, Jiang X, Morgan DG, Heuser JE, Wang X, Akey CW. Three-dimensional structure of the apoptosome: Implications for assembly, procaspase-9 binding, and activation. Mol Cell 2002; 9: 423–432. 
43. Ghavami S, Shojaei S, Yeganeh B, Ande SR, Jangamreddy JR, Mehrpour M, et al. Autophagy and apoptosis dysfunction in neurodegenerative disorders. Prog Neurobiol 2014; 112: 24–49. 
44. Agostini M, Tucci P, Melino G. Cell death pathology: Perspective for human diseases. Biochem Biophys Res Commun 2011; 414: 451–455. 
45. Bachis A, Cruz MI, Nosheny RL, Mocchetti I. Chronic unpredictable stress promotes neuronal apoptosis in the cerebral cortex. Neurosci Lett 2008; 442: 104–108. 
46. Fan C, Song Q, Wang P, Li Y, Yang M, Yu SY. Neuroprotective effects of ginsenoside-rg1 against depression-like behaviors via suppressing glial activation, synaptic deficits, and neuronal apoptosis in rats. Front Immunol 2018; 9: 1-19. 
47. McCully JD, Cowan DB, Pacak CA, Toumpoulis IK, Dayalan H, Levitsky S. Injection of isolated mitochondria during early reperfusion for cardioprotection. Am J Physiol Hear Circ Physiol 2009; 296: 94–105. 
48. McCully JD, Cowan DB, Emani SM, del Nido PJ. Mitochondrial transplantation: From animal models to clinical use in humans. Mitochondrion 2017; 34: 127–134. 
49. Ma H, Jiang T, Tang W, Ma Z, Pu K, Xu F, et al. Transplantation of platelet-derived mitochondria alleviates cognitive impairment and mitochondrial dysfunction in db/db mice. Clin Sci 2020; 134: 2161–2175. 
50. Xie Q, Zeng J, Zheng Y, Li T, Ren J, Chen K, et al. Mitochondrial transplantation attenuates cerebral ischemia-reperfusion injury: Possible involvement of mitochondrial component separation. Braun R, editor. Oxid Med Cell Longev 2021; 2021: 1–21.