Neuroprotective effect of L-deprenyl on the expression level of the Mst1 gene and inhibition of apoptosis in rat-model spinal cord injury

Document Type : Original Article


1 Department of Anatomy, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran

2 Department of Genetics and Molecular Medicine, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran


Objective(s): After primary tissue damage as a result of spinal cord injury (SCI), there is a period of secondary damage, which includes several cellular and inflammatory biochemical cascades. As a novel pro-apoptotic kinase, Mst1 (serine/threonine kinase 4) promotes programmed cell death in an inflammatory disease model. This study aimed to evaluate Mst1 gene expression levels in rats with spinal cord injury treated with L- deprenyl. 
Materials and Methods: The rats were divided into control (contusion), laminectomy, sham-operated (contused rats received 1 ml normal saline intraperitoneal), and treatment (contused rats received 5 mg/kg of L-deprenyl intraperitoneal; once a day for 7 days). The BBB (Basso, Beattie, and Bresnahan) scales were performed to assess motor function following SCI. Rats were sacrificed 28 days after SCI and the spinal cord lesion area was removed. Apoptosis and cavity formation in the spinal cord were determined by H&E staining and TUNEL assay, respectively. The mRNA levels of the Mst1, Nrf2, Bcl-2, and PGC1α genes were analyzed using real-time quantitative PCR.
Results: The results showed significant improvement in motor function in the L- deprenyl group compared with the untreated group. Histological analysis showed a significant reduction in the number of tunnel-positive cells after injection of L-deprenyl, as well as a decrease in the volume of the cavity. In addition, L-deprenyl treatment increased the expression of the Nrf2, Bcl-2, and PGC1α genes, while reducing the expression of the Mst1 gene in the spinal nerves. 
Conclusion: These results suggest that L-deprenyl is a promising treatment for spinal cord injury.


1. Shah M, Peterson C, Yilmaz E, Halalmeh DR, Moisi M. Current
advancements in the management of spinal cord injury: A comprehensive review of literature. Surg Neurol Int 2020; 11:2-7.
2. Lee JY, Chung H, Yoo YS, Oh YJ, Oh TH, Park S, et al. Inhibition of apoptotic cell death by ghrelin improves functional recovery after spinal cord injury. Endocrinology 2010; 151:3815-3826.
3. Cinar B, Fang PK, Lutchman M, Di Vizio D, Adam RM, Pavlova N, et al. The pro-apoptotic kinase Mst1 and its caspase cleavage products are direct inhibitors of Akt1. EMBO J 2007; 26:4523-4534.
4. Rawat SJ, Chernoff J. Regulation of mammalian Ste20 (Mst) kinases. Trends Biochem Sci 2015; 40:149-156.
5. Huot P, Fox SH, Brotchie JM. Monoamine reuptake inhibitors in Parkinson’s disease. Parkinsons Dis 2015; 2015:609428.
6. Naoi M, Maruyama W, Yagi K, Youdim M. Anti-apoptotic function of L-(-)deprenyl (Selegiline) and related compounds. Neurobiology (Bp) 2000; 8:69-80.
7. Naoi M, Maruyama W, Inaba-Hasegawa K. Revelation in the neuroprotective functions of rasagiline and selegiline: the induction of distinct genes by different mechanisms. Expert Review of Neurotherapeutics 2013; 13:671-684.
8. Nikfar A, Abdanipour A, Gholinejad M. Anti-apoptotic effect of selegiline as monoamine oxidase inhibitor on rat hippocampus derived neural stem cells in oxidative stress condition. J Adv Med Biomed Res 2017; 25:41-56.
9. David JA, Rifkin WJ, Rabbani PS, Ceradini DJ. The Nrf2/Keap1/ARE Pathway and Oxidative Stress as a Therapeutic Target in Type II Diabetes Mellitus. J Diabetes Res 2017; 2017:15.
10. Koundouros N, Poulogiannis G. Phosphoinositide 3-kinase/akt signaling and redox metabolism in cancer. Front Oncol 2018; 8:160-169.
11. Murakami S, Motohashi H. Roles of Nrf2 in cell proliferation and differentiation. Free Radic Biol Med 2015; 88:168-178.
12. Basso DM, Beattie MS, Bresnahan JC. Graded histological and locomotor outcomes after spinal cord contusion using the NYU weight-drop device versus transection. Exp Neurol 1996; 139:244-256.
13. Michel RP, Cruz-Orive LM. Application of the Cavalieri principle and vertical sections method to lung: estimation of volume and pleural surface area. J Microsc 1988; 150:117-136.
14. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25:402-408.
15. Mosley YC, HogenEsch H. Selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination. BMC Res Notes 2017; 10:689-696.
16. Zhang N, Yin Y, Xu SJ, Wu YP, Chen WS. Inflammation & apoptosis in spinal cord injury. Indian J Med Res 2012; 135:287-296.
17. Zhou X, He X, Ren Y. Function of microglia and macrophages in secondary damage after spinal cord injury. Neural Regen Res 2014; 9:1787-1795.
18. Inaba-Hasegawa K, Shamoto-Nagai M, Maruyama W, Naoi M. Type B and A monoamine oxidase and their inhibitors regulate the gene expression of Bcl-2 and neurotrophic factors in human glioblastoma U118MG cells: different signal pathways for neuroprotection by selegiline and rasagiline. J Neural Transm (Vienna) 2017; 124:1055-1066.
19. Tatton WG, Wadia JS, Ju WY, Chalmers-Redman RM, Tatton NA. (-)-Deprenyl reduces neuronal apoptosis and facilitates neuronal outgrowth by altering protein synthesis without inhibiting monoamine oxidase. J Neural Transm Suppl 1996; 48:45-59.
20. Abdanipour A, Jafari Anarkooli I, Shokri S, Ghorbanlou M, Bayati V, Nejatbakhsh R. Neuroprotective effects of selegiline on rat neural stem cells treated with hydrogen peroxide. Biomed Rep 2018; 8:41-46.
21. Maruyama W, Naoi M. Neuroprotection by (-)-deprenyl and related compounds. Mech Ageing Dev 1999; 111:189-200.
22. Unal I, Gursoy-Ozdemir Y, Bolay H, Soylemezoglu F, Saribas O, Dalkara T. Chronic daily administration of selegiline and EGb 761 increases brain’s resistance to ischemia in mice. Brain Res 2001; 917:174-181.
23. Magyar K, Szende B. (-)-Deprenyl, a selective MAO-B inhibitor, with apoptotic and anti-apoptotic properties. Neurotoxicology 2004; 25:233-242.
24. Xu W, Chi L, Xu R, Ke Y, Luo C, Cai J, et al. Increased production of reactive oxygen species contributes to motor neuron death in a compression mouse model of spinal cord injury. Spinal Cord 2005; 43:204-213.
25. Zhang N, Yin Y, Xu SJ, Wu YP, Chen WS. Inflammation & apoptosis in spinal cord injury. Indian J Med Res 2012; 135:287-296.
26. Dulin JN, Karoly ED, Wang Y, Strobel HW, Grill RJ. Licofelone modulates neuroinflammation and attenuates mechanical hypersensitivity in the chronic phase of spinal cord injury. J Neurosci 2013; 33:652-664.
27. Czabotar PE, Lessene G, Strasser A, Adams JM. Control of apoptosis by the BCL-2 protein family: implications for physiology and therapy. Nat Rev Mol Cell Biol 2014; 15:49-63.
28. Yuan F, Xie Q, Wu J, Bai Y, Mao B, Dong Y, et al. MST1 promotes apoptosis through regulating Sirt1-dependent p53 deacetylation. J Biol Chem 2011; 286:6940-6945.
29. Lee JY, Kim HS, Choi HY, Oh TH, Ju BG, Yune TY. Valproic acid attenuates blood–spinal cord barrier disruption by inhibiting matrix metalloprotease‐9 activity and improves functional recovery after spinal cord injury. J Neurochem 2012; 121:818-829.
30. Yuan Z, Lehtinen MK, Merlo P, Villén J, Gygi S, Bonni A. Regulation of neuronal cell death by MST1-FOXO1 signaling. J Biol Chem 2009; 284:11285-11292.
31. Lehtinen MK, Yuan Z, Boag PR, Yang Y, Villén J, Becker EB, et al. A conserved MST-FOXO signaling pathway mediates oxidative-stress responses and extends life span. Cell 2006; 125:987-1001.
32. Watabe M, Kakeya H, Onose R, Osada H. Activation of MST/Krs and c-Jun N-terminal kinases by different signaling pathways during cytotrienin A-induced apoptosis. J Biol Chem 2000; 275:8766-8771.
33. Cheung WL, Ajiro K, Samejima K, Kloc M, Cheung P, Mizzen CA, et al. Apoptotic phosphorylation of histone H2B is mediated by mammalian sterile twenty kinase. Cell 2003; 113:507-517.
34. Ardestani A, Paroni F, Azizi Z, Kaur S, Khobragade V, Yuan T, et al. MST1 is a key regulator of beta cell apoptosis and dysfunction in diabetes. Nat Med 2014; 20:385-397.
35. Zhang M, Tao W, Yuan Z, Liu Y. Mst-1 deficiency promotes post-traumatic spinal motor neuron survival via enhancement of autophagy flux. J Neurochem 2017; 143:244-256.
36. Zhang M, Tao W, Yuan Z, Liu Y. Mst‐1 deficiency promotes post‐traumatic spinal motor neuron survival via enhancement of autophagy flux. J Neurochem 2017; 143:244-256.
37. Lee JK, Shin JH, Hwang SG, Gwag BJ, McKee AC, Lee J, et al. MST1 functions as a key modulator of neurodegeneration in a mouse model of ALS. Proceedings of the National Academy of Sciences 2013; 110:12066-12071.
38. Xiao L, Chen D, Hu P, Wu J, Liu W, Zhao Y, et al. The c-Abl-MST1 signaling pathway mediates oxidative stress-induced neuronal cell death. J Neurosci 2011; 31:9611-9619.
39. Shang Y, Yan Y, Chen B, Zhang J, Zhang T. Over‐expressed MST1 impaired spatial memory via disturbing neural oscillation patterns in mice. Genes, Brain and Behavior 2020:e12678.
40. Anilkumar U, Prehn JH. Anti-apoptotic BCL-2 family proteins in acute neural injury. Front Cell Neurosci 2014; 8:281-286.
41. Zhang W, Cheng L, Hou Y, Si M, Zhao YP, Nie L. Plumbagin Protects Against Spinal Cord Injury-induced Oxidative Stress and Inflammation in Wistar Rats through Nrf-2 Upregulation. Drug Res (Stuttg) 2015; 65:495-499.
42. McBride HM, Neuspiel M, Wasiak S. Mitochondria: more than just a powerhouse. Current biology 2006; 16:R551-R560.
43. Lin J, Handschin C, Spiegelman BM. Metabolic control through the PGC-1 family of transcription coactivators. Cell metabolism 2005; 1:361-370.
44. Scarpulla RC. Nuclear control of respiratory chain expression by nuclear respiratory factors and PGC-1-related coactivator. Annals of the New York Academy of Sciences 2008; 1147:321-334.
45. Tummala KS, Kottakis F, Bardeesy N. NRF2: Translating the Redox Code. Trends Mol Med 2016; 22:829-831.
46. Kobayashi A, Kang MI, Okawa H, Ohtsuji M, Zenke Y, Chiba T, et al. Oxidative stress sensor Keap1 functions as an adaptor for Cul3-based E3 ligase to regulate proteasomal degradation of Nrf2. Mol Cell Biol 2004; 24:7130-7139.
47. Nakaso K, Nakamura C, Sato H, Imamura K, Takeshima T, Nakashima K. Novel cytoprotective mechanism of anti-parkinsonian drug deprenyl: PI3K and Nrf2-derived induction of antioxidative proteins. Biochem Biophys Res Commun 2006; 339:915-922.
48. Xiao H, Lv F, Xu W, Zhang L, Jing P, Cao X. Deprenyl prevents MPP(+)-induced oxidative damage in PC12 cells by the upregulation of Nrf2-mediated NQO1 expression through the activation of PI3K/Akt and Erk. Toxicology 2011; 290:286-294.
49. Wahdan SA, Tadros MG, Khalifa AE. Antioxidant and antiapoptotic actions of selegiline protect against 3-NP-induced neurotoxicity in rats. Naunyn Schmiedebergs Arch Pharmacol 2017; 390:905-917.
50. Inaba-Hasegawa K, Akao Y, Maruyama W, Naoi M. Type A monoamine oxidase is associated with induction of neuroprotective Bcl-2 by rasagiline, an inhibitor of type B monoamine oxidase. J Neural Transm (Vienna) 2012; 119:405-414.
51. Lin HI, Lee YJ, Chen BF, Tsai MC, Lu JL, Chou CJ, et al. Involvement of Bcl-2 family, cytochrome c and caspase 3 in induction of apoptosis by beauvericin in human non-small cell lung cancer cells. Cancer Lett 2005; 230:248-259.
52. Chiou SH, Ku HH, Tsai TH, Lin HL, Chen LH, Chien CS, et al. Moclobemide upregulated Bcl-2 expression and induced neural stem cell differentiation into serotoninergic neuron via extracellular-regulated kinase pathway. Br J Pharmacol 2006; 148:587-598.
53. Abdanipour A, Tiraihi T, Noori-Zadeh A, Majdi A, Gosaili R. Evaluation of lovastatin effects on expression of anti-apoptotic Nrf2 and PGC-1alpha genes in neural stem cells treated with hydrogen peroxide. Mol Neurobiol 2014; 49:1364-1372.