The Effects of D-aspartate on Neurosteroids, Neurosteroid Receptors, and Inflammatory Mediators in Experimental Autoimmune Encephalomyelitis

Author(s): Mahdi Goudarzvand, Yaser Panahi, Reza Yazdani, Hosein Miladi, Saeed Tahmasebi, Amin Sherafat, Sanaz Afraei, Kosar Abouhamzeh, Mahnaz Jamee, Kawthar Jasim Mohammad Rida Al-Hussieni, Hamed Mohammadi, Ali Mohebbi, Nikoo Hossein-Khannazer, Majid Zaki-Dizaji, Maria Maddalena Di Fiore, Antimo D'Aniello, Gholamreza Azizi*

Journal Name: Endocrine, Metabolic & Immune Disorders - Drug Targets
Formerly Current Drug Targets - Immune, Endocrine & Metabolic Disorders

Volume 19 , Issue 3 , 2019

Become EABM
Become Reviewer
Call for Editor

Graphical Abstract:


Objective: Experimental autoimmune encephalomyelitis (EAE) is a widely used model for multiple sclerosis. The present study has been designed to compare the efficiencies of oral and intraperitoneal (IP) administration of D-aspartate (D-Asp) on the onset and severity of EAE, the production of neurosteroids, and the expression of neurosteroid receptors and inflammatory mediators in the brain of EAE mice.

Methods: In this study, EAE was induced in C57BL/6 mice treated with D-Asp orally (D-Asp-Oral) or by IP injection (D-Asp-IP). On the 20th day, brains (cerebrums) and cerebellums of mice were evaluated by histological analyses. The brains of mice were analyzed for: 1) Neurosteroid (Progesterone, Testosterone, 17β-estradiol) concentrations; 2) gene expressions of cytokines and neurosteroid receptors by reverse transcription polymerase chain reaction, and 3) quantitative determination of D-Asp using liquid chromatography-tandem mass spectrometry. Further, some inflammatory cytokines and matrix metalloproteinase-2 (MMP-2) were identified in the mouse serum using enzyme-linked immunosorbent assay kits.

Results: Our findings demonstrated that after D-Asp was administered, it was taken up and accumulated within the brain. Further, IP injection of D-Asp had more beneficial effects on EAE severity than oral gavage. The concentration of the testosterone and 17β-estradiol in D-Asp-IP group was significantly higher than that of the control group. There were no significant differences in the gene expression of cytokine and neurosteroid receptors between control, D-Asp-IP, and D-Asp-Oral groups. However, IP treatment with D-Asp significantly reduced C-C motif chemokine ligand 2 and MMP-2 serum levels compared to control mice.

Conclusion: IP injection of D-Asp had more beneficial effects on EAE severity, neurosteroid induction and reduction of inflammatory mediators than oral gavage.

Keywords: Experimental autoimmune encephalomyelitis, matrix metalloproteinase-2, D-aspartate, neurosteroids, TNF-α, IL-6, CCL-2.

Roozbeh, M.; Mohammadpour, H.; Azizi, G.; Ghobadzadeh, S.; Mirshafiey, A. The potential role of iNKT cells in experimental allergic encephalitis and multiple sclerosis. Immunopharmacol. Immunotoxicol., 2014, 36(2), 105-113.
Fard, N.A.; Azizi, G.; Mirshafiey, A. The potential role of T helper cell 22 and IL-22 in immunopathogenesis of multiple sclerosis. Innov. Clin. Neurosci., 2016, 13(7-8), 30-36.
Azizi, G.; Haidari, M.R.; Khorramizadeh, M.; Naddafi, F.; Sadria, R.; Javanbakht, M.H.; Sedaghat, R.; Tofighi Zavareh, F.; Mirshafiey, A. Effects of imatinib mesylate in mouse models of multiple sclerosis and in vitro determinants. Iran. J. Allergy Asthma Immunol., 2014, 13(3), 198-206.
Azizi, G.; Goudarzvand, M.; Afraei, S.; Sedaghat, R.; Mirshafiey, A. Therapeutic effects of dasatinib in mouse model of multiple sclerosis. Immunopharmacol. Immunotoxicol., 2015, 37(3), 287-294.
Afraei, S.; D’Aniello, A.; Sedaghat, R.; Ekhtiari, P.; Azizi, G.; Tabrizian, N.; Magliozzi, L.; Aghazadeh, Z.; Mirshafiey, A. Therapeutic effects of D-aspartate in a mouse model of multiple sclerosis. J. Food Drug Anal., 2017, 25(3), 699-708.
Goudarzvand, M.; Afraei, S.; Yaslianifard, S.; Ghiasy, S.; Sadri, G.; Kalvandi, M.; Alinia, T.; Mohebbi, A.; Yazdani, R.; Azarian, S.K.; Mirshafiey, A.; Azizi, G. Hydroxycitric acid ameliorates inflammation and oxidative stress in mouse models of multiple sclerosis. Neural Regen. Res., 2016, 11(10), 1610-1616.
Azizi, G. Mirshafiey, A.; Imatinib mesylate: An innovation in treatment of autoimmune diseases. Recent Pat. Inflamm. Allergy Drug Discov., 2013, 7(3), 259-267.
Afraei, S.; Azizi, G.; Zargar, S.J.; Sedaghat, R.; Mirshafiey, A. New therapeutic approach by G2013 in experimental model of multiple sclerosis. Acta Neurol. Belg., 2015, 115(3), 259-266.
D’Aniello, A.; Di Fiore, M.M.; Fisher, G.H.; Milone, A.; Seleni, A.; D’Aniello, S.; Perna, A.F.; Ingrosso, D. Occurrence of D-aspartic acid and N-methyl-D-aspartic acid in rat neuroendocrine tissues and their role in the modulation of luteinizing hormone and growth hormone release. FASEB J., 2000, 14(5), 699-714.
D’Aniello, S.; Somorjai, I.; Garcia-Fernandez, J.; Topo, E.; D’Aniello, A. D-Aspartic acid is a novel endogenous neurotransmitter. FASEB J., 2011, 25(3), 1014-1027.
Di Fiore, M.M.; Santillo, A.; Chieffi Baccari, G. Current knowledge of D-aspartate in glandular tissues. Amino Acids, 2014, 46(8), 1805-1818.
Santillo, A.; Pinelli, C.; Burrone, L.; Chieffi Baccari, G.; Di Fiore, M.M. D-Aspartic acid implication in the modulation of frog brain sex steroid levels. Gen. Comp. Endocrinol., 2013, 181, 72-76.
Burrone, L.; Santillo, A.; Pinelli, C.; Chieffi Baccari, G.; Di Fiore, M.M. Induced synthesis of P450 aromatase and 17beta-estradiol by D-aspartate in frog brain. J. Exp. Biol., 2012, 215(Pt 20), 3559-3565.
Di Fiore, M.M.; Santillo, A.; Falvo, S.; Longobardi, S.; Chieffi Baccari, G. Molecular mechanisms elicited by d-aspartate in leydig cells and spermatogonia. Int. J. Mol. Sci., 2016, 17(7), pii: E1127.
Santillo, A.; Falvo, S.; Chieffi, P.; Burrone, L.; Chieffi Baccari, G.; Longobardi, S.; Di Fiore, M.M. D-aspartate affects NMDA receptor-extracellular signal-regulated kinase pathway and upregulates androgen receptor expression in the rat testis. Theriogenology, 2014, 81(5), 744-751.
Di Fiore, M.M.; Santillo, A.; Falvo, S.; Chieffi Baccari, G.; Venditti, M.; Di Giacomo Russo, F.; Lispi, M.; D’Aniello, A. Sex hormone levels in the brain of d-aspartate-treated rats. C. R. Biol., 2018, 341(1), 9-15.
Falvo, S.; Di Fiore, M.M.; Burrone, L.; Chieffi Baccari, G.; Longobardi, S.; Santillo, A. Androgen and oestrogen modulation by D-aspartate in rat epididymis. Reprod. Fertil. Dev., 2016, 28(12), 1865-1872.
Harris, V.K.; Bell, L.; Langan, R.A.; Tuddenham, J.; Landy, M.; Sadiq, S.A. Fetuin-A deficiency protects mice from Experimental Autoimmune Encephalomyelitis (EAE) and correlates with altered innate immune response. PLoS One, 2017, 12(4), e0175575.
Fontanarosa, C.; Pane, F.; Sepe, N.; Pinto, G.; Trifuoggi, M.; Squillace, M.; Errico, F.; Usiello, A.; Pucci, P.; Amoresano, A. Quantitative determination of free D-Asp, L-Asp and N-methyl-D-aspartate in mouse brain tissues by chiral separation and multiple reaction monitoring tandem mass spectrometry. PLoS One, 2017, 12(6), e0179748.
Naddafi, F.; Reza Haidari, M.; Azizi, G.; Sedaghat, R.; Mirshafiey, A. Novel therapeutic approach by nicotine in experimental model of multiple sclerosis. Innov. Clin. Neurosci., 2013, 10(4), 20-25.
Errico, F.; Napolitano, F.; Squillace, M.; Vitucci, D.; Blasi, G.; de Bartolomeis, A.; Bertolino, A.; D’Aniello, A.; Usiello, A. Decreased levels of D-aspartate and NMDA in the prefrontal cortex and striatum of patients with schizophrenia. J. Psychiatr. Res., 2013, 47(10), 1432-1437.
Nuzzo, T.; Sacchi, S.; Errico, F.; Keller, S.; Palumbo, O.; Florio, E.; Punzo, D.; Napolitano, F.; Copetti, M.; Carella, M.; Chiariotti, L.; Bertolino, A.; Pollegioni, L.; Usiello, A. Decreased free daspartate levels are linked to enhanced d-aspartate oxidase activity in the dorsolateral prefrontal cortex of schizophrenia patients. NPJ Schizophr., 2017, 3, 16.
Errico, F.; Nistico, R.; Di Giorgio, A.; Squillace, M.; Vitucci, D.; Galbusera, A.; Piccinin, S.; Mango, D.; Fazio, L.; Middei, S.; Trizio, S.; Mercuri, N.B.; Teule, M.A.; Centonze, D.; Gozzi, A.; Blasi, G.; Bertolino, A.; Usiello, A. Free D-aspartate regulates neuronal dendritic morphology, synaptic plasticity, gray matter volume and brain activity in mammals. Transl. Psychiatry, 2014, 4, e417.
Azizi, G.; Khannazer, N.; Mirshafiey, A. The potential role of chemokines in alzheimer’s disease pathogenesis. Am. J. Alzheimers Dis. Other Demen., 2014, 29(5), 415-425.
Yousefi, B.; Jadidi-Niaragh, F.; Azizi, G.; Hajighasemi, F.; Mirshafiey, A. The role of leukotrienes in immunopathogenesis of rheumatoid arthritis. Mod. Rheumatol., 2014, 24(2), 225-235.
Javanbakht, M.H.; Sadria, R.; Djalali, M.; Derakhshanian, H.; Hosseinzadeh, P.; Zarei, M.; Azizi, G.; Sedaghat, R.; Mirshafiey, A. Soy protein and genistein improves renal antioxidant status in experimental nephrotic syndrome. Nefrologia, 2014, 34(4), 483-490.
Mirshafiey, A.; Simhag, A.; El Rouby, N.M.; Azizi, G. T-helper 22 cells as a new player in chronic inflammatory skin disorders. Int. J. Dermatol., 2015, 54(8), 880-888.
Yazdani, R.; Sharifi, M.; Shirvan, A.S.; Azizi, G.; Ganjalikhani-Hakemi, M. Characteristics of innate lymphoid cells (ILCs) and their role in immunological disorders (an update). Cell. Immunol., 2015, 298(1-2), 66-76.
Beck, J.; Rondot, P.; Catinot, L.; Falcoff, E.; Kirchner, H.; Wietzerbin, J. Increased production of interferon gamma and tumor necrosis factor precedes clinical manifestation in multiple sclerosis: do cytokines trigger off exacerbations? Acta Neurol. Scand., 1988, 78(4), 318-323.
Erta, M.; Quintana, A.; Hidalgo, J. Interleukin-6, a major cytokine in the central nervous system. Int. J. Biol. Sci., 2012, 8(9), 1254-1266.
Stelmasiak, Z.; Koziol-Montewka, M.; Dobosz, B.; Rejdak, K.; Bartosik-Psujek, H.; Mitosek-Szewczyk, K.; Belniak-Legiec, E. Interleukin-6 concentration in serum and cerebrospinal fluid in multiple sclerosis patients. Med. Sci. Monit., 2000, 6(6), 1104-1108.
Rey, M.; Coirini, H. Synthetic neurosteroids on brain protection. Neural Regen. Res., 2015, 10(1), 17-21.
Takao, T.; Flint, N.; Lee, L.; Ying, X.; Merrill, J.; Chandross, K.J. 17beta-estradiol protects oligodendrocytes from cytotoxicity induced cell death. J. Neurochem., 2004, 89(3), 660-673.
Noorbakhsh, F.; Baker, G.B.; Power, C. Allopregnanolone and neuroinflammation: a focus on multiple sclerosis. Front. Cell. Neurosci., 2014, 8, 134.
D’Aniello, A.; Di Cosmo, A.; Di Cristo, C.; Annunziato, L.; Petrucelli, L.; Fisher, G. Involvement of D-aspartic acid in the synthesis of testosterone in rat testes. Life Sci., 1996, 59(2), 97-104.
Azcoitia, I.; Leonelli, E.; Magnaghi, V.; Veiga, S.; Garcia-Segura, L.M.; Melcangi, R.C. Progesterone and its derivatives dihydroprogesterone and tetrahydroprogesterone reduce myelin fiber morphological abnormalities and myelin fiber loss in the sciatic nerve of aged rats. Neurobiol. Aging, 2003, 24(6), 853-860.

Rights & PermissionsPrintExport Cite as

Article Details

Year: 2019
Published on: 04 October, 2018
Page: [316 - 325]
Pages: 10
DOI: 10.2174/1871530318666181005093459
Price: $65

Article Metrics

PDF: 27