Volume 30, Issue 2 (June 2026)
Physiol Pharmacol 2026, 30(2): 168-184 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Mohammadzadeh M, Abdolmaleki Z, Karimi Goudarzi A, Bolandi S M. Combined effects of SH-SY5Y–derived exosomes
and glutamic acid on neuroinflammation and
functional recovery after MCAO in rats. Physiol Pharmacol 2026; 30 (2) :168-184
URL: http://ppj.phypha.ir/article-1-2462-en.html
URL: http://ppj.phypha.ir/article-1-2462-en.html
Abstract: (1488 Views)
Introduction: Ischemic stroke and brain trauma are significant causes of death and disability in the West. Inflammasomes induce pro-inflammatory cytokine production in cerebral ischemia, thereby driving inflammation. The current study investigates the effects of glutamic acid, N-methyl-D-aspartate receptor agonist, and SH-SY5Y Neuroblastoma-Derived Exosomes in the rat model of cerebral ischemia.
Methods: Sixty adult male Wistar rats were randomly assigned to five groups. After MCAO, rats were treated with glutamic acid, SH-SY5Y neuroblastoma-derived exosomes, and both. Neurological deficits were assessed using the Garcia scale 7 days after ischemia. Neuronal cell death was evaluated using cresyl violet staining. We examined gene and protein levels of BDNF, NLRP3, and NMDAR in brain tissue using real-time PCR and immunohistochemistry. TTC staining was used to determine cerebral infarct volume in brain tissue, and ELISA tests to measure IL-1β and IL-18 levels.
Results: Cresyl violet staining demonstrated that glutamic acid+exosomes significantly reduced neuronal degeneration. Garcia’s test results showed that glutamic acid, independently or in combination with exosomes, significantly improved sensory and motor functions. The results of IHC and Real-Time PCR clearly indicated that the expression of the BDNF gene and protein, as well as the NMDAR gene, increased, whereas the expression of the NLRP3 gene and protein, and the Caspase-1 protein, decreased. ELISA results showed a significant drop in IL-1β and IL-18 levels in MCAO groups treated with glutamic acid, exosomes, or both.
Conclusion: The use of glutamic acid with SH-SY5Y neuroblastoma-derived exosomes exerts neuroprotective benefits by enhancing neurotrophic factors and diminishing apoptosis in the MCAO model.
Methods: Sixty adult male Wistar rats were randomly assigned to five groups. After MCAO, rats were treated with glutamic acid, SH-SY5Y neuroblastoma-derived exosomes, and both. Neurological deficits were assessed using the Garcia scale 7 days after ischemia. Neuronal cell death was evaluated using cresyl violet staining. We examined gene and protein levels of BDNF, NLRP3, and NMDAR in brain tissue using real-time PCR and immunohistochemistry. TTC staining was used to determine cerebral infarct volume in brain tissue, and ELISA tests to measure IL-1β and IL-18 levels.
Results: Cresyl violet staining demonstrated that glutamic acid+exosomes significantly reduced neuronal degeneration. Garcia’s test results showed that glutamic acid, independently or in combination with exosomes, significantly improved sensory and motor functions. The results of IHC and Real-Time PCR clearly indicated that the expression of the BDNF gene and protein, as well as the NMDAR gene, increased, whereas the expression of the NLRP3 gene and protein, and the Caspase-1 protein, decreased. ELISA results showed a significant drop in IL-1β and IL-18 levels in MCAO groups treated with glutamic acid, exosomes, or both.
Conclusion: The use of glutamic acid with SH-SY5Y neuroblastoma-derived exosomes exerts neuroprotective benefits by enhancing neurotrophic factors and diminishing apoptosis in the MCAO model.
Type of Manuscript: Experimental research article |
Subject:
Neurophysiology/Pharmacology
References
1. Belov Kirdajova D, Kriska J, Tureckova J, Anderova M. Ischemia-triggered glutamate excitotoxicity from the perspective of glial cells. Frontiers in Cellular Neuroscience 2020; 14: 51. [DOI:10.3389/fncel.2020.00051]
2. Bordi F, Pietra C, Ziviani L, Reggiani A. The glycine antagonist GV150526 protects somatosensory evoked potentials and reduces the infarct area in the MCAo model of focal ischemia in the rat. Experimental Neurology 1997; 145: 425-433. [DOI:10.1006/exnr.1997.6442]
3. Cai Z, Lin S, Rhodes P G. Neuroprotective effects of N-acetylaspartylglutamate in a neonatal rat model of hypoxia-ischemia. European Journal of Pharmacology 2002; 437: 139-145. [DOI:10.1016/S0014-2999(02)01289-X]
4. Casals J B, Pieri N C, Feitosa M L, Ercolin A, Roballo K, Barreto R S, et al. The use of animal models for stroke research: a review. Comparative Medicine 2011; 61: 305-313.
5. Cavelle D, Greenamyre J T. Selective vulnerability of the CA1 region of hippocampus to the indirect excitotoxic effects of malonic acid. Neuroscience Letters 1995; 192: 29-32. [DOI:10.1016/0304-3940(95)11600-2]
6. Chen L, Deng H, Cui H, Fang J, Zuo Z, Deng J, et al. Inflammatory responses and inflammation-associated diseases in organs. Oncotarget 2018; 9: 7204-7218. [DOI:10.18632/oncotarget.23208]
7. Cheng N T, Kim A S. Intravenous thrombolysis for acute ischemic stroke within 3 hours versus between 3 and 4.5 hours of symptom onset. The Neurohospitalist 2015; 5: 101-109. [DOI:10.1177/1941874415583116]
8. Constantino L C, Tasca C I, Boeck C R. The role of NMDA receptors in the development of brain resistance through pre-and postconditioning. Aging and Disease 2014; 5: 430.
9. Dhawan J, Benveniste H, Luo Z, Nawrocky M, Smith S D, Biegon A. A new look at glutamate and ischemia: NMDA agonist improves long-term functional outcome in a rat model of stroke. Future Neurology 2011; 6: 823-834. [DOI:10.2217/fnl.11.55]
10. Fauré J, Lachenal G, Court M, Hirrlinger J, Chatellard-Causse C, Blot B, et al. Exosomes are released by cultured cortical neurones. Molecular and Cellular Neuroscience 2006; 31: 642-648. [DOI:10.1016/j.mcn.2005.12.003]
11. Feigin V L, Stark B A, Johnson C O, Roth G A, Bisignano C, Abady G G, et al. Global, regional, and national burden of stroke and its risk factors, 1990-2019: a systematic analysis for the Global Burden of Disease Study 2019. The Lancet Neurology 2021; 20: 795-820. [DOI:10.1016/S1474-4422(21)00252-0]
12. Feng Y-S, Tan Z-X, Wang M-M, Xing Y, Dong F, Zhang F. Inhibition of NLRP3 inflammasome: a prospective target for the treatment of ischemic stroke. Frontiers in Cellular Neuroscience 2020; 14: 155. [DOI:10.3389/fncel.2020.00155]
13. Franke M, Bieber M, Kraft P, Weber A N R, Stoll G, Schuhmann M K. The NLRP3 inflammasome drives inflammation in ischemia/reperfusion injury after transient middle cerebral artery occlusion in mice. Brain, Behavior, and Immunity 2021; 92: 223-233. [DOI:10.1016/j.bbi.2020.12.009]
14. Garcia J H, Wagner S, Liu K-F, Hu X-j. Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rats: statistical validation. Stroke 1995; 26: 627-635. [DOI:10.1161/01.STR.26.4.627]
15. Gupta Y K, Chauhan A. Potential of immunosuppressive agents in cerebral ischaemia. The Indian Journal of Medical Research 2011; 133: 15.
16. Huang L, Hua L, Zhang X. The Exosomal MicroRNA Profile Is Responsible for the Mesenchymal Stromal Cell Transplantation-Induced Improvement of Functional Recovery after Stroke in Rats. Neuroimmunomodulation 2022; 29: 151-160. [DOI:10.1159/000518637]
17. Huo L, Du X, Li X, Liu S, Xu Y. The emerging role of neural cell-derived exosomes in intercellular communication in health and neurodegenerative diseases. Frontiers in Neuroscience 2021; 15: 738442. [DOI:10.3389/fnins.2021.738442]
18. Ismael S, Zhao L, Nasoohi S, Ishrat T. Inhibition of the NLRP3-inflammasome as a potential approach for neuroprotection after stroke. Scientific Reports 2018; 8: 5971. [DOI:10.1038/s41598-018-24350-x]
19. Jeanblanc J, Coune F, Botia B, Naassila M. Brain-derived neurotrophic factor mediates the suppression of alcohol self-administration by memantine. Addiction Biology 2014; 19: 758-769. [DOI:10.1111/adb.12039]
20. Kotlęga D, Peda B, Zembroń-Łacny A, Gołąb-Janowska M, Nowacki P. The role of brain-derived neurotrophic factor and its single nucleotide polymorphisms in stroke patients. Neurologia i Neurochirurgia Polska 2017; 51: 240-246. [DOI:10.1016/j.pjnns.2017.02.008]
21. Kuriakose D, Xiao Z. Pathophysiology and treatment of stroke: present status and future perspectives. International Journal of Molecular Sciences 2020; 21: 7609. [DOI:10.3390/ijms21207609]
22. Latz E, Xiao T. Stutz AJNRI. Activation and Regulation of the Inflammasomes. Nature Reviews Immunology 2013; 13: 397-411. [DOI:10.1038/nri3452]
23. Lau C G, Zukin R S. NMDA receptor trafficking in synaptic plasticity and neuropsychiatric disorders. Nature Reviews Neuroscience 2007; 8: 413-426. [DOI:10.1038/nrn2153]
24. Lee B-R, Kim J-H, Choi E-S, Cho J-H, Kim E. Effect of young exosomes injected in aged mice. International Journal of Nanomedicine 2018: 5335-5345. [DOI:10.2147/IJN.S170680]
25. Liu H-D, Li W, Chen Z-R, Hu Y-C, Zhang D-D, Shen W, et al. Expression of the NLRP3 inflammasome in cerebral cortex after traumatic brain injury in a rat model. Neurochemical Research 2013; 38: 2072-2083. [DOI:10.1007/s11064-013-1115-z]
26. Liu Q, Li S, Dupuy A, Mai H l, Sailliet N, Logé C, et al. Exosomes as new biomarkers and drug delivery tools for the prevention and treatment of various diseases: current perspectives. International Journal of Molecular Sciences 2021a; 22: 7763. [DOI:10.3390/ijms22157763]
27. Liu X, Fan L, Li J, Bai Z, Wang Y, Liu Y, et al. Mailuoning oral liquid attenuates convalescent cerebral ischemia by inhibiting AMPK/mTOR-associated apoptosis and promoting CREB/BDNF-mediated neuroprotection. Journal of Ethnopharmacology 2023; 317: 116731. [DOI:10.1016/j.jep.2023.116731]
28. Liu X, Zhang M, Liu H, Zhu R, He H, Zhou Y, et al. Bone marrow mesenchymal stem cell-derived exosomes attenuate cerebral ischemia-reperfusion injury-induced neuroinflammation and pyroptosis by modulating microglia M1/M2 phenotypes. Experimental Neurology 2021b; 341: 113700. [DOI:10.1016/j.expneurol.2021.113700]
29. Lu X, Dong J, Zheng D, Li X, Ding D, Xu H. Reperfusion combined with intraarterial administration of resveratrol-loaded nanoparticles improved cerebral ischemia-reperfusion injury in rats. Nanomedicine: Nanotechnology, Biology and Medicine 2020; 28: 102208. [DOI:10.1016/j.nano.2020.102208]
30. Muir K W, Lees K R. Clinical experience with excitatory amino acid antagonist drugs. Stroke 1995; 26: 503-513. [DOI:10.1161/01.STR.26.3.503]
31. Nazarian S, Abdolmaleki Z, Torfeh A, Shirazi Beheshtiha S H. Mesenchymal stem cells with modafinil (gold nanoparticles) significantly improves neurological deficits in rats after middle cerebral artery occlusion. Experimental Brain Research 2020; 238: 2589-2601. [DOI:10.1007/s00221-020-05913-9]
32. Olivares D, K Deshpande V, Shi Y, K Lahiri D, H Greig N, T Rogers J, et al. N-methyl D-aspartate (NMDA) receptor antagonists and memantine treatment for Alzheimer’s disease, vascular dementia and Parkinson’s disease. Current Alzheimer Research 2012; 9: 746-758. [DOI:10.2174/156720512801322564]
33. Pocock J M, Kettenmann H. Neurotransmitter receptors on microglia. Trends in Neurosciences 2007; 30: 527-535. [DOI:10.1016/j.tins.2007.07.007]
34. Rajani V, Sengar A S, Salter M W. Tripartite signalling by NMDA receptors. Molecular Brain 2020; 13: 23. [DOI:10.1186/s13041-020-0563-z]
35. Ramanathan M, Abdul K K, Justin A. Low dose of l-glutamic acid attenuated the neurological dysfunctions and excitotoxicity in bilateral common carotid artery occluded mice. Behavioural Pharmacology 2016; 27: 615-622. [DOI:10.1097/FBP.0000000000000256]
36. Safakheil M, Safakheil H. The effect of exosomes derived from bone marrow stem cells in combination with rosuvastatin on functional recovery and neuroprotection in rats after ischemic stroke. Journal of Molecular Neuroscience 2020; 70: 724-737. [DOI:10.1007/s12031-020-01483-1]
37. Salaudeen M A, Bello N, Danraka R N, Ammani M L. Understanding the pathophysiology of ischemic stroke: the basis of current therapies and opportunity for new ones. Biomolecules 2024; 14: 305. [DOI:10.3390/biom14030305]
38. Salazar-Colocho P, Del Río J, Frechilla D. Neuroprotective effects of serotonin 5-HT1A receptor activation against ischemic cell damage in gerbil hippocampus: Involvement of NMDA receptor NR1 subunit and BDNF. Brain Research 2008; 1199: 159-166. [DOI:10.1016/j.brainres.2007.12.032]
39. Shi J, Gao W, Shao F. Pyroptosis: Gasdermin-Mediated Programmed Necrotic Cell Death. Trends in Biochemical Sciences 2017; 42: 245-254. [DOI:10.1016/j.tibs.2016.10.004]
40. Song L, Pei L, Yao S, Wu Y, Shang Y. NLRP3 inflammasome in neurological diseases, from functions to therapies. Frontiers in Cellular Neuroscience 2017; 11: 63. [DOI:10.3389/fncel.2017.00063]
41. Tian T, Cao L, He C, Ye Q, Liang R, You W, et al. Targeted delivery of neural progenitor cell-derived extracellular vesicles for anti-inflammation after cerebral ischemia. Theranostics 2021; 11: 6507-6521. [DOI:10.7150/thno.56367]
42. Van Hemelrijck A, Hachimi-Idrissi S, Sarre S, Ebinger G, Michotte Y. Neuroprotective effect of N-acetyl-aspartyl-glutamate in combination with mild hypothermia in the endothelin-1 rat model of focal cerebral ischaemia. Journal of Neurochemistry 2005; 95: 1287-1297. [DOI:10.1111/j.1471-4159.2005.03450.x]
43. Vanni I, Alama A, Grossi F, Dal Bello M G, Coco S. Exosomes: a new horizon in lung cancer. Drug Discovery Today 2017; 22: 927-936. [DOI:10.1016/j.drudis.2017.03.004]
44. Wei H, Chen J, Wang S, Fu F, Zhu X, Wu C, et al. A nanodrug consisting of doxorubicin and exosome derived from mesenchymal stem cells for osteosarcoma treatment in vitro. International Journal of Nanomedicine 2019: 8603-8610. [DOI:10.2147/IJN.S218988]
45. Wu Q J, Tymianski M. Targeting NMDA receptors in stroke: new hope in neuroprotection. Molecular Brain 2018; 11: 1-14. [DOI:10.1186/s13041-018-0357-8]
46. Zhang J, Li S, Li L, Li M, Guo C, Yao J, et al. Exosome and exosomal microRNA: trafficking, sorting, and function. Genomics, Proteomics & Bioinformatics 2015; 13: 17-24. [DOI:10.1016/j.gpb.2015.02.001]
47. Zhang X, Sai B, Wang F, Wang L, Wang Y, Zheng L, et al. Hypoxic BMSC-derived exosomal miRNAs promote metastasis of lung cancer cells via STAT3-induced EMT. Molecular Cancer 2019; 18: 1-15. [DOI:10.1186/s12943-019-0959-5]
48. Zhang Y, Yu J, Liu J, Liu H, Li J. Effects of stem cell-derived exosomes on neuronal apoptosis and inflammatory cytokines in rats with cerebral ischemia-reperfusion injury via PI3K/AKT pathway-mediated mitochondrial apoptosis. Immunopharmacology and Immunotoxicology 2021; 43: 731-740. [DOI:10.1080/08923973.2021.1976794]
49. Zheng C, Qiao Z H, Hou M Z, Liu N N, Fu B, Ding R, et al. GLYX-13, a NMDA receptor glycine-site functional partial agonist, attenuates cerebral ischemia injury in vivo and vitro by differential modulations of nmda receptors subunit components at different post-ischemia stage in mice. Frontiers in Aging Neuroscience 2017; 9: 186. [DOI:10.3389/fnagi.2017.00186]
50. Zhu H, Hu S, Li Y, Sun Y, Xiong X, Hu X, et al. Interleukins and Ischemic Stroke. Frontiers in Immunology 2022; 13: 828447. [DOI:10.3389/fimmu.2022.82844]
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
