Amelioration of amyloid beta (Aβ1-40) neurotoxicity by administration of silibinin; a behavioral and biochemical assessment

Document Type : Original Article


1 Department of Anatomy, School of Medicine, Ilam University of Medical Sciences, lam Iran

2 Biotechnology and Medicinal Plants Research Center, School of Medicine, Ilam University of Medical Sciences, Ilam, Iran

3 Department of Physiology, School of Medicine, Ilam University of Medical Sciences, Ilam Iran



Objective(s): Alzheimer’s disease (AD), the most common cause of dementia, is one of the leading causes of morbidity and death in the world. Currently, treatment mostly used to slow down the disease progression. Herbal remedies are considered by many in the community as a natural and safe treatment with fewer side effects. Silibinin, the active ingredient of Silybum marionum, has anti-oxidant, neurotrophic and neuroprotective characteristics. Therefore, here, the effect of different doses of Silibinin extract on oxidative stress and expression of neurotrophic factors was investigated. 
Materials and Methods: Forty eight male Wistar rats were randomly divided into sham, lesion; Aβ1-40 injection, lesion-treatment; Aβ1-40 injection followed by different doses of silibinin (50, 100, 200 mg / kg) through gavage and lesion-vehicle group; Aβ1-40 injection + vehicle of silibinin. Morris water Maze (MWM) was done 28 days after the last treatment. Hippocampal tissue was removed for biochemical analysis. Production of nitric oxide (NO) and reactive oxygen species (ROS), expression of BDNF/VEGF and cell viability were measured using Griess, fluorimetry, Western blotting and MTT techniques.  
Results: Different concentrations of silibinin improved behavioral performance in animals. Higher doses of Silibinin could improve memory and learning function through MWM. Also, increasing the concentration of silibinin resulted in decreased ROS and NO production in a dose-dependent manner.
Conclusion: Consequently, silibinin may act as a potential candidate for alleviating symptoms of AD.


Main Subjects

1.    Mansergh FC, Wride MA, Rancourt DE. Neurons from stem cells: implications for understanding nervous system development and repair. Biochem Cell Biol 2000;78:613-268.
2.    C Qiu, M Kivipelto, E Von Strauss. Epidemiology of Alzheimer’s disease: occurrence, determinants, and strategies toward intervention. Dialogues Clin Neurosci 2009; 11: 111–128.
3.    Stix G. Alzheimer’s: Forestalling the darkness. Sci Am 2010;302:50-57.
4.    Zhang H, Ma Q, Zhang Yw, Xu H. Proteolytic processing of Alzheimer’s β‐amyloid precursor protein. J Neurochem 2012;120:9-21.
5.    Cunnane SC, Plourde M, Pifferi F, Bégin M, Féart C, Barberger-Gateau P. Fish, docosahexaenoic acid and Alzheimer’s disease. Prog Lipid Res 2009;48:239-256.
6.    Schmitt-Schillig S, Schaffer S, Weber C, Eckert G, Muller W. Flavonoids and the aging brain. J Physiol Pharmacol 2005;56:23-36.
7.    Liebl MP, Kaya AM, Tenzer S, Mittenzwei R, Koziollek-Drechsler I, Schild H, et al. Dimerization of visinin-like protein 1 is regulated by oxidative stress and calcium and is a pathological hallmark of amyotrophic lateral sclerosis. Free Radical Bio Med 2014;72:41-54.
8.    Bossy-Wetzel E, Schwarzenbacher R, Lipton SA. Molecular pathways to neurodegeneration. Nat Med 2004;10:S2-S9.
9.    Purdy P, Ericsson S, Dodson R, Sternes K, Garner D. Effects of the flavonoids, silibinin and catechin, on the motility of extended cooled caprine sperm. Small Ruminant Res 2004;55:239-243.
10.    Schulz V, Hänsel R, Tyler VE. Rational phytotherapy: a physician’s guide to herbal medicine: Psychology Press; 2001.
11.    Mateen S, Tyagi A, Agarwal C, Singh RP, Agarwal R. Silibinin inhibits human nonsmall cell lung cancer cell growth through cell‐cycle arrest by modulating expression and function of key cell‐cycle regulators. Mol Carcinog 2010;49:247-258.
12.    Baluchnejadmojarad T, Roghani M, Mafakheri M. Neuroprotective effect of silymarin in 6-hydroxydopamine hemi-parkinsonian rat: involvement of estrogen receptors and oxidative stress. Neurosci Lett 2010;480:206-210.
13.    Geed M, Garabadu D, Ahmad A, Krishnamurthy S. Silibinin pretreatment attenuates biochemical and behavioral changes induced by intrastriatal MPP+ injection in rats. Pharmacol Biochem Be 2014;117:92-103.
14.    Hou Y-C, Liou K-T, Chern C-M, Wang Y-H, Liao J-F, Chang S, et al. Preventive effect of silymarin in cerebral ischemia–reperfusion-induced brain injury in rats possibly through impairing NF-κB and STAT-1 activation. Phytomedicine 2010;17:963-73.
15.    Jangra A, Kasbe P, Pandey SN, Dwivedi S, Gurjar SS, Kwatra M, et al. Hesperidin and silibinin ameliorate aluminum-induced neurotoxicity: modulation of anti-oxidants and inflammatory cytokines level in mice hippocampus. Biol Trace Elem Res 2015;168:462-471.
16.    Paxinos G WC. The Rat Brain in Stereotaxic Coordinates. Academic, New York 1998.
17.    Bagheri M, Rezakhani A, Nyström S, Turkina MV, Roghani M, Hammarström P, et al. Amyloid beta1-40-induced astrogliosis and the effect of genistein treatment in rat: a three-dimensional confocal morphometric and proteomic study. PloS One 2013;8:e76526.
18.    Bagheri M, Rezakhani A, Roghani M, Joghataei MT, Mohseni SJJ. Protocol for three-dimensional confocal morphometric analysis of astrocytes. J Vis Exp 2015:e53113.
19.    Shankar GM, Walsh DMJMn. Alzheimer’s disease: synaptic dysfunction and Aβ. Mol Neurodegener 2009;4:1-13.
20.    Carrillo-Mora P, Luna R, Colin-Barenque L. Amyloid beta: multiple mechanisms of toxicity and only some protective effects? Oxid Med Cell Longev 2014;2014:795375. 
21.    dos Santos VV, Santos DB, Lach G, Rodrigues ALS, Farina M, De Lima TC, et al. Neuropeptide Y (NPY) prevents depressive-like behavior, spatial memory deficits and oxidative stress following amyloid-β (Aβ1–40) administration in mice. Behav Brain Res 2013;244:107-115.
22.    Raza SS, Khan MM, Ashafaq M, Ahmad A, Khuwaja G, Khan A, et al. Silymarin protects neurons from oxidative stress associated damages in focal cerebral ischemia: a behavioral, biochemical and immunohistological study in Wistar rats. J Neurol Sci 2011;309:45-54.
23.    Shen L, Liu L, Li X-Y, Ji H-F. Regulation of gut microbiota in Alzheimer’s disease mice by silibinin and silymarin and their pharmacological implications. Appl Microbiol Biot 2019;103:7141-7149.
24.    Lu P, Mamiya T, Lu L, Mouri A, Niwa M, Hiramatsu M, et al. Silibinin attenuates amyloid β25–35 peptide-induced memory impairments: implication of inducible nitric-oxide synthase and tumor necrosis factor-α in mice. J Pharmacol Exp Ther 2009;331:319-326.
25.    Jung UJ, Jeon MT, Choi MS, Kim SR. Silibinin attenuates MPP(+)-induced neurotoxicity in the substantia nigra in vivo. J Med Food 2014;17:599-605. 
26.    Lima Giacobbo B, Doorduin J, Klein HC, Dierckx R, Bromberg E, de Vries EFJ. Brain-Derived Neurotrophic Factor in Brain Disorders: Focus on Neuroinflammation. Mol Neurobiol 2019;56:3295-3312. 
27.    Nowacka M, Obuchowicz E. BDNF and VEGF in the pathogenesis of stress-induced affective diseases: an insight from experimental studies. Pharmacol Rep 2013;65:535-546. 
28.    Song X, Liu B, Cui L, Zhou B, Liu W, Xu F, et al. Silibinin ameliorates anxiety/depression-like behaviors in amyloid β-treated rats by upregulating BDNF/TrkB pathway and attenuating autophagy in hippocampus. Physiol Behav 2017;179:487-493.
29.    Yön B, Belviranlı M, Okudan NJJoB. The effect of silymarin supplementation on cognitive impairment induced by diabetes in rats. J Basic Clin Physiol Pharmacol 2019;30:1-9.