Neem Leaf Supplement Ameliorates Depressive Like Behaviour in Alzheimer’s Disease Model in Adult Male Wistar Rats

Adejoke Elizabeth Memudu *

Department of Anatomy Faculty of Basic Medical Science Edo State University Uzairue, Edo State, Nigeria.

Fatima Ayinda Anzaku

Department of Anatomy, Faculty of Basic Medical Sciences, Bingham University; P.M.B 005, Karu, Nasarawa State, Nigeria.

Grace Mchibuma Jibaniya

Department of Anatomy, Faculty of Basic Medical Sciences, Bingham University; P.M.B 005, Karu, Nasarawa State, Nigeria and Department of Anatomy, Faculty of Basic Medical Sciences, Federal University Wukari, Taraba State, Nigeria.

Rukky Precious Adanike

Harbor UCLA Medical Center, Torrance, CA, United States of America.

*Author to whom correspondence should be addressed.


Aim of the Study: Currently, reports linked neuropathological changes in Alzheimer’s Disease (AD) to be a risk factor in depression. There is a need to develop natural therapeutics, with strong antioxidant property like Neem leaf, to avert AD’s neuropathological and behavioural changes using animal model induced by neurotoxin aluminium chloride. This study is aimed at ascertaining depressive like disorders in AD model while evaluating mechanism through which Neem averts AD neurodegeneration in the fronto-cerebellar cortex and possible depressive like behaviour.

Methodology: Twenty (20) adult male Wistar rats used were grouped (n=5) viz: control group (A), neurodegenerative model given aluminium (B), 200mg/kg Neem leaf supplement (C) and 200mg/kg Neem leaf treated AD model (D). Neurobehavioural changes for reward memory and depressive like behaviour were evaluated using Y-maze (open arm reward test) and the tail suspension test (TST).  The frontal and cerebellar cortices were excised, fixed and processed for  Haematoxylin and Eosin stain (H and E), Cresyl fast violet (CFV) stain for Nissl bodies and astrocytes immunohistochemistry using glial fibrillary acidic protein (GFAP). Behavioural test data were analyzed using ANOVA and test for significance done @ p<0.05.

Results: Aluminum induced neurodegeneration similar to AD pathology characterized by loss of neurons, chromatolysis, proliferation of astrocytes and decline in cognitive function in addition to exhibiting depressive like behaviour seen in a decrease number of reward arm entries, increase in time spent to reach reward arm and immobility time in the TST attributed to loss of cognitive function relative to loss of neurons integrity in the fronto-cerebellar cortex. However, neem leaf supplementation mitigates against AD model neuropathological and neurobehavioural  presentations resulting in an improved neurocognition, neuron survival or repair, decline in astrocytes proliferation and decline in depressive like behavior as compared to Aluminum induced AD model.

Conclusion: Neem leaf alleviates depressive like behaviour associated with neurocognitive impairment through the interaction between neuron-astrocytes to protect neurons against aluminum induced neuroinflamamation and strengthens neural circuit for cognitive function.

Keywords: Depression, astrocytes, chromatolysis, neem leaf supplement, neuroinflammation, Alzheimer's disease, memory impairment, fronto-cerebellar cortex

How to Cite

Memudu , A. E., Anzaku , F. A., Jibaniya , G. M., & Adanike , R. P. (2024). Neem Leaf Supplement Ameliorates Depressive Like Behaviour in Alzheimer’s Disease Model in Adult Male Wistar Rats. Asian Journal of Research in Medical and Pharmaceutical Sciences, 13(1), 31–46.


Download data is not yet available.


Pan PY, Bölte S, Kaur P, Jamil S, Jonsson U. Neurological disorders in autism: A systematic review and meta-analysis. Autism. 2020;20:1362361320951370.

Mogle J, Hill N.L, Bhargava S et al. Memory complaints and depressive symptoms over time: A construct-level replication analysis. BMC Geriatr. 2020; 20:57

Maramis MM, Mahajudin MS, Khotib J. Impaired cognitive flexibility and working memory precedes depression: A rat model to study depression. Neuropsychobiology. 2021;80:225–233

Valverde AP, Camargo A, Rodrigues ALS. Agmatine as a novel candidate for rapid-onset antidepressant response. World J Psychiatr. 2021;11(11):981-996 [PMID: 34888168 DOI: 10.5498/wjp.v11.i11.981]

Lee TH, Choi JH, Shin BC, Shin HE, Son HR. Correlation between dementia and depressive disorder. KJFP. 2018;8:327-330.

Fang Y, Ding X, Zhang Y et al. Fluoxetine inhibited the activation of A1 reactive astrocyte in a mouse model of major depressive disorder through astrocytic 5-HT2BR/β-arrestin2 pathway. J Neuroinflammation. 2022;19:23.

Yuan C, Chen H, Wang Y, Schneider JA, Willett WC, Morris MC. Dietary carotenoids related to risk of incident Alzheimer Dementia (AD) and brain AD neuropathology: A community-based cohort of older adults. Am J Clin Nutr. 2021;113(1):200–208. Available:

Ellouze I, Sheffler J, Nagpal R, Arjmandi B. Dietary patterns and alzheimer’s disease: An updated review linking nutrition to neuroscience. Nutrients. 2023;15(14): 3204. Available:

Yu O, Jung B, Go H, et al. Association between dementia and depression: a retrospective study using the Korean National Health Insurance Service-National Sample Cohort database BMJ Open 2020; 10:e034924.

Lanza C, Sejunaite K, Steindel C, Scholz I, Riepe MW. Cognitive profiles in persons with depressive disorder and Alzheimer’s disease. Brain Communications. 2020;2 (2):fcaa206.

Solch RJ, Aigbogun JO, Voyiadjis AG, Talkington GM, Darensbourg RM, O’Connell S, Pickett KM, Perez SR, Maraganore DM. Mediterranean diet adherence, gut microbiota, and Alzheimer’s or Parkinson’s disease risk: A systematic review. J. Neurol. Sci. 2022; 434:120166.

Exely Vickers T. Journal of medical case reports. “Elevated brain aluminium and early onset of alzheimer’s disease in an individual occupationally exposed to aluminium: A Case Report. 2014;8:41.

Akinrinade ID, Memudu AE, Ogundele OM. Fluoride and Aluminium disturb neuronal morphology, transport functions, cholinesterase, lysosomal, and cell cycle activity. Pathophysiology 2015;(22):105 -115.

Memudu Adejoke Elizabeth, Pantong Samson and Osahon Roli Itohan. Histomorphological evaluations on the frontal cortex extrapyramidal cell layer following administration of N-Acetyl cysteine in aluminum induced neuro-degeneration rat model. Metab Brain Dis. 2020;35:829–839. Available:

Brenner S. Aluminum may mediate Alzheimer's disease through liver toxicity, with aberrant hepatic synthesis of ceruloplasmin and ATPase7B, the resultant excess free copper causing brain oxidation, beta-amyloid aggregation and Alzheimer disease. Med Hypotheses. 2013; 80(3):326-327

Belovicova K, Bogi E, Csatlosova K, Dubovicky M. Animal tests for anxiety-like and depression-like behavior in rats. Interdisciplinary Toxicology. 2017;10(1): 40–43. Available:

Raghavendra M, Achara SB, Rituparna M, Kumar S. Role of aqueous extract of Aindica leaves in an experimental model of Alzheimer’s disease in rats. International Journal of Applied Basic Medical Research. 2013l;3(1):37-47. DOI:10.4103/2229-516x.112239

Fakunle PB, Ajibade AJ, Oyewo EB, Adeyemi OH. A study of some effects of aqueous extract of neem (Azadirachta Indica) Leaves on the lead acetate induced neurotoxicity in the superficial layers of superior colliculus of adult wistar rats ( Rattus norvegicus );2012.

Sujarwo W/, Keim AP, Caneva G, Toniolo C, Nicoletti M. Ethnobotanical uses of neem leaves in Bali (indonesia) and the Indian subcontinent in relation with historical background and phytochemical properties. Journal of Ethnophharmacology. 2016;189:186-93. DOI: 10.1016/j.jep.2016.05.014

Sinha JD, Nandaha DSK, Jaiswal N, Vasudeva A, Prabha TS, Singh PU. Antibacterial effect of Azadirachtaindica (Neem) or Curcuma longa (turmeric) against enterococcus faecal;is compared with that of 5% sodium hypochlorite or 2% chlorohexidine in vitro. Bull Tokoyo Dent Coll. 2017;58(2):103-109. DOI:10.2209/tdcpublication.2015-0029

Datta A, Fernado LN, Grun IU, Kwasniewski MT. Comparison of two absorbentbased de-bittering procedures for neem a Tea-effect on polyphrnols, anti-oxidant capacity, color and volatile profile. Plant Foods Human Nutrition. 2017;72 (1):88-95. DOI:10.1007/s11130-016-0505-9

Sáiz-Vázquez O, Gracia-García P, Ubillos-Landa S, Puente-Martínez A, Casado-Yusta S, Olaya B, Santabárbara J. Depression as a risk factor for alzheimer’s disease: A systematic review of longitudinal meta-analyses. J. Clin. Med. 2021;10:1809

National research council. Guide for the care and use of laboratory animals. (8th Edition). Washington, DC: The National Academies Press.2011;123-124.

Buraimoh AA, Ojo SA, Hambolu JO. Effects of oral administration of aluminum chloride on the histology of the hippocampus of wistar rats. Current Research Journal of Biological Sciences. 2011;3(5):509-515. ISSN: 2041-0778.

American psychological association, animal research and ethics in 2012: American Psychological Association (APA) guidelines for ethical conduct in the care and use of animals;2012.

National research council. Guidelines for the care and use of mammals in neuroscience and behavioral research. Washington, DC: The National Academies Press;2003.

Onaolapo OJ, Abiodun OR, Akanji OO, Mosaku TJ, Onaolapo AY. Elevated plus maze and Y-maze behavioral effects of subchronic, oral low dose monosodium glutamate in swiss albino mice. IOSR Journal of Pharmacy and Biological Sciences. 2012;3(4):21-27. ISSN2278-3008

Cryan JF, Mombereau C, Vassout A. The tail suspension test as a model for assessing antidepressant activity: Review of pharmacological and genetic studies in mice. Neurosci Biobehav Rev2005;29:571-625

Can A, Dao DT, Terrillion CE, Piantadosi SC, Bhat S, Gould TD. The tail suspension test. Journal of Visualized Experiments .2012;(59):3769. DOI: 10.3791/3769

Carbone L, Elizabeth TC, Elizabeth MY, Diana BB, Krista AL, John MP, Jamie AA, Youngho S, Anisha DG, James DW. Assessing cervical dislocation as a human Euthansaia Method in Mice. J.Am.Assoc Lab Anim Sci. 2012;51(3):352-356

AVMA guidelines for the euthanasia of animals, 2013. AVMA Guidelines for the euthanasia of animals: 2013 Edition American Veterinary Medical Associationon Euthanasia. AVMA Guidelines; 2007

Bancroft JD, Gamble M. Theory and practice of Histological techniques. 6th Edition London Churchill Livingstone: 2008;374 – 375

Bancroft John D, Christopher Layton, Kim Suvarna S. Bancroft's theory and practice ofhistological techniques Oxford]: Churchill livingstone elsevier: 7th ed;2013.

Paxinos and Watson Brain Stereotaxis mapping (Paxinos and Watson, 2006)::; Paxinos, george, and Charles Watson. The rat brain in stereotaxic coordinates: hard cover edition. Access Online via Elsevier; 2006.

Giri D. Histopathology. “Hematoxylin and Eosin staining: principle, procedure and interpretation;2015.

Memudu AE, Adewumi. Alpha-lipoic acid ameliorates scopolamine-induced memory deficit and neurodegeneration in the cerebello-hippocampal cortex. Metabolic Brain Disease. 2021;36(7): 1729–1745.

DOI: 10.1007/s11011-021-00720-9

Buriamoh AA, Ojo SA. Effects of aluminum chloride exposure on the body weight of wistar rats. Annals of Biological Sciences. 2014;2(2):66-73. ISSN:2348-1927.

Berk M, Dean OM, Cotton SM, Jeavons S, Tanious M, Kohlmann K, Hewitt K, Moss K, Allwang C, Schapkaitz I, et al. The efficacy of adjunctive nacetylcysteine in major depressive disorder: A double-blind, randomized, placebo-controlled trial. J Clin Psychiatry. 2014;75(6):628–636.

Giancarlo A, Alessandra A, Giovanna B, Giulio V, Marina C, Luisa B, Francesco S NAcetylcysteine as an antioxidant and disulphide breaking agent: the reasons why, Free Radical Research. 2018; 52(7):751-762. DOI: 10.1080/10715762.2018.1468564

Schmutterer. The Neem tree and other Melaiaceous plants. Sources of unique natural products for integrated past management, medicine industry & other purposes; 2014.

Alzohairy MA. Therapeutics role of Azadirachtaindica (Neem) and their active constituent in disease prevention and treatment. Evidence-Based Complementary and Alternative Medicine. eCAM 2016:7382506. DOI:10.1155/2016/7382506

Blaylock R. Aluminum induced immunoexcitotoxicity in neurodevelopmental and neurode-generative disorders. Current Inorganic Chemistry. 2012;2(1):46-53. DOI:10.2174/1877944111202010046.

Jana M, Dasgupta S. Ghorpade A, Pahan K. Neuroimmune pharmacology- astrocytes, oligodendrites, and schwann cells. 2017;69-88.

Kunyu Li, Jiatong Li, Jialin Zheng, Song Qin. Reactive astrocytes in Neurodegenerative Diseases. Aging and disease. 2019;10(3):664-675

Paumier A, Boisseau S, Jacquier-Sarlin M, Pernet-Gallay K, Buisson AA, Albrieux M. Astrocyte–neuron interplay is critical for Alzheimer's disease pathogenesis and is rescued by TRPA1 channel blockade, Brain. 2022;145 (1):388–405

Dolotov OV, Inozemtseva LS, Myasoedov NF, Grivennikov IA. Stress-induced depression and alzheimer’s disease: Focus on astrocytes. International Journal of Molecular Sciences. 2022;23(9): 4999

Zhou X, Qian X, Li X, Fan Y, Liping W, Jie T. Astrocyte, a promising target for mood disorder interventions. Frontiers in Molecular Neuroscience. 2019;12.

Liao Y, Xing Q, Li Q et al. Astrocytes in depression and Alzheimer’s disease. Front. Med. 2021;15:829–841. Available:

Kamel ES, Mostafa N. The Egyptian Journal of Histology. Effects of aluminum chloride on the hippocampus of adult rats and the possible protective role of Nigella sativa: A histological and immune-histochemical study.