Moringa oleifera lam. Leaf Extract Preserves Spatial Memory and Hippocampal Microstructure in Aluminium Chloride-Induced Neurotoxicity in Adult Wistar Rats
DOI:
https://doi.org/10.47081/njn2025.16.1/004Keywords:
Aluminium, Hippocampus, Immunohistochemistry, Moringa oleifera, HistologyAbstract
Aluminium is a widely available and subtly consumed metal, whose actions on human health include neurotoxicity and cognitive decline, among others. It is important to protect against these aluminium effects through antioxidants, as aluminium utilizes the oxidative pathway to exert its effect. Due to the abundance of antioxidants in Moringa oleifera (M. oleifera), this study investigated its neuroprotective potential against aluminium chloride (AlCl3)-induced hippocampal intoxication in Wistar rats. The phytochemical screening and toxicity (LD50) of M. oleifera leaf ethanol extract (MO) were evaluated. Thirty adult male Wistar rats (150–220 g) were then assigned into six groups (n = 5): control, AlCl3 (100 mg/kg), M. oleifera low dose (MOLD, 250 mg/kg), M. oleifera high dose (MOHD, 1,000 mg/kg), concomitant AlCl3 + MOLD, and AlCl3 + MOHD. These administrations were oral and lasted for 21 days. On day 22, spontaneous alternation behaviour (SAB) was tested in the T-maze, the animals sacrificed, and the brains processed for histology and immunoreactivity. The phytochemicals of MO included flavonoids, phenols, alkaloids, saponins, and tannins as the major constituents, while its LD50 was greater than 5,000 mg/kg. There was significantly (p < 0.05) less SAB, hippocampal cornu ammonis (CA) 3 chromatolysis, and decreased neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expressions in the AlCl3 group. Concomitant treatment with MOLD and MOHD did not significantly improve SAB, and NSE and GFAP expressions, but preserved Nissl distribution. MO protected the hippocampus against AlCl3 intoxication by improving SAB, and modulating Nissl distribution, NSE, and GFAP expressions, which supports its antioxidant potential.
Downloads
References
Abdel-Rahman Mohamed, A., Metwally, M.M.M., Khalil, S.R., Salem, G.A. and Ali, H.A. (2019) Moringa oleifera extract attenuates the CoCl2 induced hypoxia of rat’s brain: Expression pattern of HIF-1α, NF-kB, MAO and EPO. Biomed Pharmacother. 109:1688-1697. doi:10.1016/j.bio pha.2018.11.019
Abijo, A.Z., Adeeyo, O.O., Komolafe, O.A., Saka, O.S., and Abodunrin, V.K. (1970) Effects of Moringa oleifera on the developing cerebrum of young wistar rats. Anat J Afr. 8(1):1336-1341. doi:10.4314/aja.v8i1.182596
Adedapo, A.A., Falayi, O.O. and Oyagbemi, A.A. (2015) Evaluation of the analgesic, anti-inflammatory, anti-oxidant, phytochemical and toxicological properties of the methanolic leaf extract of commercially processed Moringa oleifera in some laboratory animals. J Basic Clin Physiol Pharmacol. 26:491-499. doi:10.1515/jbcpp-2014-0105
Ademiluyi, A.O., Aladeselu, O.H., Oboh, G. and Boligon, A.A. (2018) Drying alters the phenolic constituents, antioxidant properties, α‐amylase, and α‐glucosidase inhibitory properties of Moringa (Moringa oleifera) leaf. Food Sci Nutr. 6:2123–2133. doi:10.1002/fsn3.770
Adighije, N.K., Ekerete, I.A. and Ekong, M. (2020) Effect of Moringa oleifera Lam. leaf extract against aluminium chloride induced hippocampal histology and serum enzyme activities in adult Wistar rats. Int J Med Surg Sci, 61-74. doi:10.32457/ijmss.v7i2.503
Akpanyung, E.O., Nwaokonko, D.U., Ekong, M.B. and Ekpo, M.M. (2018) Evaluation of the protective effect of moringa oleifera leaf extract against aluminium induced liver damage in male albino Wistar rats. Int J Sci. 4:20-30. doi:10.18483/ijSci.1520
Ashfaq, M., Basra, S.M.A. and Ashfaq, U. (2012) Moringa: A Miracle Plant for Agro-forestry. J Agric Soc Sci. 8(3):115-122.
Ballaz, S.J., and Rebec, G.V. (2019) Neurobiology of vitamin C: Expanding the focus from antioxidant to endogenous neuromodulator. Pharmacol Res. 146: 104321. doi:10.1016/j.phrs.2019.104321
Bharosay, A., Bharosay, V.V., Varma, M., Saxena, K., Sodani, A., and Saxena, R. (2012) Correlation of brain biomarker neuron specific enolase (NSE) with degree of disability and neurological worsening in cerebrovascular stroke. Ind J Clin Biochem 27:186–190. doi:10.1007/ s12291-011-0172-9
Bondan, E.F., Martins, M.D.F.M. and Viani, F.C. (2013) Decreased astrocytic GFAP expression in streptozotocin-induced diabetes after gliotoxic lesion in the rat brainstem. Arq Bras Endocrinol Metab. 57:431–436. doi:10.1590/ S0004-27302013000600004
Bruce, R. (1985) An up-and-down procedure for acute toxicity testing. Fundam Appl Toxicol. 5:151–157. doi: 10.1016/0272-0590(85)90059-4
Bryliński, Ł., Kostelecka, K., Woliński, F., Duda, P., Góra, J., Granat, M., et al. (2023) Aluminium in the human brain: routes of penetration, toxicity, and resulting complications. Int J Mol Sci. 24:7228. doi:10.3390/ijms24087228
Butterfield, D.A. and Lange, M.L.B. (2009) Multifunctional roles of enolase in Alzheimer’s disease brain: Beyond altered glucose metabolism. Journal of Neurochemistry 111, 915–933. doi:10.1111/j.1471-4159.2009.06397.x
Byrne, J.H. (2014) From Molecules to Networks: An Introduction to Cellular and Molecular Neuroscience. 3rd ed. San Diego: Elsevier Science & Technology.
Çabuş, N., Oğuz, E., Tufan, A. and Adıgüzel, E. (2015) A histological study of toxic effects of aluminium sulfate on rat hippocampus. Biotec Histochem. 90:132–139. doi: 10.3109/10520295.2014.965277
Cho, K.S., Lim, Y., Lee, K., Lee, J., Lee, J.H., and Lee, I.-S. (2017) Terpenes from forests and human health. Toxicol Res. 33:97-106. doi:10.5487/TR.2017.33.2.097
Chu, X., Zhou, S., Sun, R., Wang, L., Xing, C., Liang, R., et al. (2018). Chrysophanol relieves cognition deficits and neuronal loss through inhibition of inflammation in diabetic mice. Neurochem Res 43:972-983. doi:10.1007/s11064-018-2503-1
Deacon, R.M.J. and Rawlins, J.N.P. (2006) T-maze alternation in the rodent. Nat Protoc 1:7–12. doi: 10.1038/nprot.2006.2
Ding, M., Haglid, K.G. and Hamberger, A. (2000) Quantitative immunochemistry on neuronal loss, reactive gliosis and BBB damage in cortex/striatum and hippocampus/amygdala after systemic kainic acid administration. Neurochem Int. 36:313-318. doi: 10.1016/S0197-0186(99)00139-4
Dinkova-Kostova, A.T. and Kostov, R.V. (2012) Glucosinolates and isothiocyanates in health and disease. Trends Mol Med 18:337–347. doi:10.1016/j.molmed.2012. 04.003
Ekong, M.B., Ekpo, M.M., Akpanyung, E.O. and Nwaokonko, D.U. (2017) Neuroprotective effect of Moringa oleifera leaf extract on aluminium-induced temporal cortical degeneration. Metab Brain Dis. 32:1437–1447. doi: 10.1007/s11011-017-0011-7
Evans, W.C. (2009) Trease and Evans Pharmacognosy., 16th Edn. London: Saunder Publisher.
Exley, C. and Vickers, T. (2014) Elevated brain aluminium and early onset alzheimer’s disease in an individual occupationally exposed to aluminium: a case report. J Med Case Reports 8:41. doi: 10.1186/1752-1947-8-41
Falowo, A.B., Mukumbo, F.E., Idamokoro, E.M., Lorenzo, J.M., Afolayan, A.J. and Muchenje, V. (2018) Multi-functional application of Moringa oleifera Lam. in nutrition and animal food products: A review. Food Res Int. 106:317-334. doi:10.1016/j.foodres.2017.12.079
Farhat, S.M., Mahboob, A., Iqbal, G. and Ahmed, T. (2017) Aluminum-induced cholinergic deficits in different brain parts and its implications on sociability and cognitive functions in mouse. Biol Trace Elem Res. 177:115-121. doi:10.1007/s12011-016-0856-3
Ferreira, P.M.P., Farias, D.F., Oliveira, J.T.D.A. and Carvalho, A.D.F.U. (2008) Moringa oleifera: Bioactive compounds and nutritional potential. Rev Nutr. 21:431-437. doi:10.1590/S1415-52732008000400007
Flora, S.J.S. and Pachauri, V. (2011) Moringa (Moringa oleifera) seed extract and the prevention of oxidative stress, In: Nuts and Seeds in Health and Disease Prevention. Elsevier. 775-785. doi:10.1016/B978-0-12-375688-6.10092-1
Fulgenzi, A., Vietti, D., and Ferrero, M.E. (2014) Aluminium involvement in neurotoxicity. BioMed Res Int. 2014:1–5. doi:10.1155/2014/758323
Guo‐Ross, S.X., Yang, E.Y., Walsh, T.J. and Bondy, S.C. (1999) Decrease of glial fibrillary acidic protein in rat frontal cortex following aluminum treatment. J Neurochem. 73:1609–1614. doi:10.1046/j.1471-4159.1999.0731609.x
Haque, A., Polcyn, R., Matzelle, D. and Banik, N.L. (2018) New insights into the role of neuron-specific enolase in neuro-inflammation, neurodegeneration, and neuroprotect ion. Brain Sci. 8:33. doi:10.3390/brainsci80 20033
Harborne, J.B. (1984). Phytochemical Methods: A Guide to Modern Techniques of Plant Analysis. Dordrecht: Springer.
Jayasree, T.M., Kishore, K.K., Rupa, L M., and Dixit, RR. (2012) Evaluation of hypoglycemic and antihyperglycemic effect of freshly prepared aqueous extract of Moringa oleifera leaves in normal and diabetic rabbits. J Chem Pharm Res. 4(1):249-253
Kaur, K., Narang, R.K. and Singh, S. (2022) AlCl3 induced learning and memory deficit in zebrafish. NeuroToxicology. 92:67-76. doi:10.1016/j.neuro.2022.07.004
Krupińska, I. (2020) Aluminium drinking water treatment residuals and their toxic impact on human health. Molecules. 25:641. doi:10.3390/molecules25030641
Kuznetsova, I.A., Areshidze, D.A. and Kozlova, M.A. (2017) The influence of different aluminium compounds on the hippocampal morphofunctional state and conditioning in mice. Toxicol Environ Health Sci. 9:215–221. doi: 10.1007/s13530-017-0323-3
Lee, K.H., Cha, M. and Lee, B.H. (2020) Neuroprotective effect of antioxidants in the brain. Int J Mol Sci. 21(19):7152. doi:10.3390/ijms21197152
Li, D., Liu, X., Liu, T., Liu, H., Tong, L., Jia, S., et al. (2020) Neurochemical regulation of the expression and function of glial fibrillary acidic protein in astrocytes. Glia. 68:878–897. doi: 10.1002/glia.23734
Lin, H., Zhu, H., Tan, J., Wang, H., Wang, Z., Li, P., et al. (2019) Comparative analysis of chemical constituents of moringa oleifera leaves from China and India by ultra-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry. Molecules 24:942. doi:10.3390/molecules24050942
Liu, C., Li, Y., Lein, P. J. and Ford, B.D. (2012) Spatiotem poral patterns of GFAP upregulation in rat brain following acute intoxication with diisopropylfluorophosphate (DFP). Curr Neurobiol. 3:90–97.
Lobo, M.G., Hounsome, N. and Hounsome, B. (2018) Biochemistry of vegetables: secondary metabolites in vegetables - terpenoids, phenolics, alkaloids, and sulfur‐containing compounds. In: Handbook of Vegetables and Vegetable Processing. Siddiq, M. and Uebersax, M.A (Eds.). Wiley. pp. 47-82. doi:10.1002/9781119098935.ch3
Lopez-Teros, V., Ford, J.L., Green, M.H., Tang, G., Grusak, M.A., Quihui-Cota, L., et al. (2017) Use of a “super-child” approach to assess the vitamin a equivalence of Moringa oleifera leaves, develop a compartmental model for vitamin a kinetics, and estimate vitamin a total body stores in young Mexican children. J Nutr. 147:2356–2363. doi:10.3945/jn.117.256974
Mahdi, H.J., Khan, N.A.K., Asmawi, M.Z.B., Mahmud, R., and A/L Murugaiyah, V. (2018) In vivo anti-arthritic and anti-nociceptive effects of ethanol extract of Moringa oleifera leaves on complete Freund’s adjuvant (CFA)-induced arthritis in rats. Integr Med Res. 7:85–94. doi: 10.1016/j.imr.2017.11.002
McFarland, G., La Joie, E., Thomas, P. and Lyons-Weiler, J. (2020) Acute exposure and chronic retention of aluminum in three vaccine schedules and effects of genetic and environmental variation. J Trace Elem Med Biol. 58:126444. doi:10.1016/j.jtemb.2019.126444
Nampoothiri, M., Kumar, N., Venkata Ramalingayya, G., Gopalan Kutty, N., Krishnadas, N., and Mallikarjuna Rao, C. (2017) Effect of insulin on spatial memory in aluminum chloride-induced dementia in rats. NeuroReport. 28:540–544. doi:10.1097/WNR.0000000000000799
National Research Council (2011) Guide for the Care and Use of Laboratory Animals. 8th Edn. Washington, D.C.: National Academies Press. doi: 10.17226/12910
Nimse, S. B., and Pal, D. (2015) Free radicals, natural antioxidants, and their reaction mechanisms. RSC Adv. 5, 27986–28006. doi:10.1039/C4RA13315C
Nogami, M., Takatsu, A. and Ishiyama, I. (1998) Immunohistochemical study of neuron-specific enolase in human brains from forensic autopsies. Forensic Science Int. 94:97-109. doi: 10.1016/S0379-0738(98)00060-7
OECD (2002) Guidance Document on Acute Oral Toxicity Testing. OECD. doi:10.1787/9789264078413-en
Polcyn, R., Capone, M., Hossain, A., Matzelle, D., Banik, N. L. and Haque, A. (2017) Neuron specific enolase is a potential target for regulating neuronal cell survival and death: implications in neurodegeneration and regeneration. Neurosciences. 4:254-257. doi:10.20517/2347-8659.2017. 59
Santhi, K. and Sengottuvel, R. (2016) Qualitative and quantitative phytochemical analysis of Moringa concanensis Nimmo. Int J Curr Microbiol App Sci. 5:633-640. doi:10.20546/ijcmas.2016.501.064
Smith, M.E. and Eng, L.F. (1987) Glial fibrillary acidic protein in chronic relapsing experimental allergic encephalomyelitis in SJL/J mice. J Neurosci Res. 18:203-208. doi:10.1002/jnr.490180129
Steffek, A.E., McCullumsmith, R.E., Haroutunian, V. and Meador-Woodruff, J.H. (2008) Cortical expression of glial fibrillary acidic protein and glutamine synthetase is decreased in schizophrenia. Schizophr Res. 103:71-82. doi:10.1016/j.schres.2008.04.032
Stohs, S.J. and Hartman, M.J. (2015) Review of the safety and efficacy of Moringa oleifera. Phytother Res. 29:796–804. doi:10.1002/ptr.5325
Suttie, A.E., Bradley, A.E., Boorman, G.A., and Leininger, J.R. (2018) Boorman’s Pathology of the Rat: Reference and Atlas. 2nd edn. London: Academic Press, Elsevier.
Suvarna, K.S., Layton, C. and Bancroft, J.D. (2019) Bancroft’s Theory and Practice of Histological Techniques., 8th edn. Elsevier. doi:10.1016/C2015-0-00143-5
Taïr, K., Kharoubi, O., Taïr, O.A., Hellal, N., Benyettou, I. and Aoues, A. (2016) Aluminium-induced acute neurotoxicity in rats: Treatment with aqueous extract of Arthrophytum (Hammada scoparia). J Acute Dis. 5:470-482. doi:10.1016/j.joad.2016.08.028
Venkatesh, K., Srikanth, L., Vengamma, B., Chandra sekhar, C., Sanjeevkumar, A., Mouleshwara Prasad, B., et al. (2013) In vitro differentiation of cultured human CD34+ cells into astrocytes. Neurol India. 61:383. doi: 10.4103/0028-3886.117615
Vergara-Jimenez, M., Almatrafi, M. and Fernandez, M. (2017) Bioactive components in Moringa oleifera leaves protect against chronic disease. Antioxidants. 6:91. doi: 10.3390/antiox6040091
Vermerris, W. and Nicholson, R. (2006) Phenolic compounds and their effects on human health. In: Phenolic Compound Biochemistry. Dordrecht: Springer. Pp.235–255. doi:10.1007/978-1-4020-5164-7_7
Villarruel-López, A., López-de La Mora, D.A., Vázquez-Paulino, O.D., Puebla-Mora, A.G., Torres-Vitela, M.R., Guerrero-Quiroz, L.A., et al. (2018) Effect of Moringa oleifera consumption on diabetic rats. BMC Complement Altern Med. 18:127. doi:10.1186/s12906-018-2180-2
Virk, S.A. and Eslick, G.D. (2015) Aluminum levels in brain, serum, and cerebrospinal fluid are higher in Alzheimer’s disease cases than in controls: A series of meta-analyses. J Alzheimers Dis. 47(3):629–638. doi:10.3233/JAD-150193
Wang, X., Cheng, D., Jiang, W. and Ma, Y. (2018) Mechanisms underlying aluminum neurotoxicity related to 14-3-3ζ protein. Toxicol Sci. 163:45–56. doi:10.1093/ toxsci/kfy021
Yang, X., Cao, Z., Zhang, J., Shao, B., Song, M., Han, Y., et al. (2018) Dendritic spine loss caused by AlCl3 is associated with inhibition of the Rac 1/cofilin signaling pathway. Environ Pollut. 243:1689–1695. doi:10.1016/j. envpol.2018.09.145
Yardimoğlu, M., İlbay, G., Dalçik, C., Dalçik, H., Sahi˙n, D. and Ateş, N. (2008) Immunocytochemistry of neuron specific enolase (NSE) in the rat brain after single and repeated epileptic seizures. Int J Neurosc. 118:981–993. doi:10.1080/00207450701769232
Yuan, C.-Y., Lee, Y.-J. and Hsu, G.-S.W. (2012) Aluminum overload increases oxidative stress in four functional brain areas of neonatal rats. J Biomed Sci. 19:51. doi: 10.1186/1423-0127-19-51
Zhang, Z., Ma, Z., Zou, W., Guo, H., Liu, M., Ma, Y., et al. (2019) The appropriate marker for astrocytes: comparing the distribution and expression of three astrocytic markers in different mouse cerebral regions. Biomed Res Int. 2019: 1-15. doi: 10.1155/2019/9605265
Zhou, J., Yang, W., Suo, D., Li, Y., Peng, L., Xu, L., et al. (2018) Moringa oleifera seed extract alleviates scopolamine-induced learning and memory impairment in mice. Front Pharmacol. 9:389. doi: 10.3389/fphar.2018. 00389
Downloads
Published
Issue
Section
License
Copyright (c) 2025 Published articles are licensed under Creative Commons Attribution CC BY 4.0, which permits use, sharing, adaptation, distribution, and reproduction in any medium or format, as long as appropriate credit is given to the original author(s) and the source(s).

This work is licensed under a Creative Commons Attribution 4.0 International License.
Copyrights for articles are retained by the authors, with first publication rights granted to the journal. Authors have rights to reuse, republish, archive, and distribute their own articles after publication. The journal/publisher is not responsible for subsequent uses of the work. This journal is licenced under a Creative Commons Attribution 4.0 License CC BY 4.0 (https://creativecommons.org/licenses/by/4.0/).