| Research Article |
Open Access |
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| Neuroprotective Effect of Curcumin with a Fixator of Absorption against
both Aluminium Neurotoxicity and Alzheimer’s Disease (Experimental
Studies in Mice) |
| Salima Douichene1*, Noureddine Djebli1, Moussa Ahmed2 and Kheira Zerrouki1 |
| 1Department of Biology, Faculty of SE & SNV, Laboratory of Micro Biology and Vegetal Biology (LMVB), Mostaganem University, Mostaganem, Algeria |
| 2Department of Biology, Faculty of Sciences, Tiaret University, Tiaret, Algeria |
| *Corresponding author: |
Salima Douichene
Department of Biology, Faculty of
SE & SNV
Laboratory of Micro Biology and Vegetal Biology (LMVB)
Mostaganem
University, Mostaganem, Algeria
E-mail: itadz@yahoo.fr |
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| Received June 13, 2012; Accepted August 10, 2012; Published August 13, 2012 |
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| Citation: Douichene S, Djebli N, Ahmed M, Zerrouki K (2012) Neuroprotective
Effect of Curcumin with a Fixator of Absorption against both Aluminium Neurotoxicity
and Alzheimer’s Disease (Experimental Studies in Mice). J Alzheimers Dis
Parkinsonism 2:107. doi:10.4172/2161-0460.1000107 |
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| Copyright: © 2012 Douichene S, et al. This is an open-access article distributed
under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the
original author and source are credited. |
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| Abstract |
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| The versatility of Aluminium chemistry will ensure its burgeoning use in the future in all manner of applications.
This very same chemistry will also ensure its increased biological availability in all biota including man. We do
not know enough about the biological chemistry of a chronic and sub-acute exposure to Aluminium to be able
to predict its impact on human health. Although the hypothesis of a link between Aluminium and Alzheimer's
Disease (AD) has been supported by several epidemiological studies, there is much controversy regarding these
findings and their interpretation. There is no doubt that the drugs discovered for neurodegenerative disease
are far from ideal, and are not more than acetyl cholinesterase inhibitors, in fact alternative medicine could be
moderate and protect nerves from neurotoxicity and AD. Curcumin is one of strong medicinal species that prove
antioxidative stress effect against reactions cascade of neurofibrillary tangles. Our aims is to clarify the role of
Curcumin with a fixative of absorption as a protective and therapeutic agent against neurodegenerative disorders
and Alzheimer's disease induced by AlCl3 on the pyramidal cells in cerebral cortex and hippocampus of albino
mice, by means of two experiences realized with a different exposition of albino mice; sub acute and chronic
exposure with a different manner (PO and IP) on Curcumin with a fixative of absorption and AlCl3. |
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| Keywords |
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| Curcumin; Neurodegenerative disorders; Alzheimer’s
disease; AlCl3 |
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| Introduction |
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| Aluminum (Al) is a common metal in the environment and one
of the most abundant in the terrestrial crust. Human exposure to Al
has been increasing over the last few decade’s [1]. This element appears
mainly in food products and in drinking water derived from both
natural sources and treatment methods Oxidative injury has been
suggested to contribute to neurodegenerative and metabolic disorders
[2]. Epidemiological studies have indicated a link between Al in
drinking water and AD and a variety of human and animal studies have
implicated learning and memory deficit after Al exposure [3,4]. |
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| Drugs currently available on the market include different inhibitors
of acetylcholine esterase and N-methyl D-aspartate (NMDA) receptor
antagonist. These drugs improve the function of still intact neurons, but
do not inhibit the ongoing degenerative process leading the neuronal
cell death [5]. |
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| Curcumin is a biological active component of turmeric (curcuma
longa) is used as a curry spice and herbal medicine for the treatment
of inflammatory conditions, cancer, AIDS, and other diseases [6].
Results show that a low dose of curcumin significantly suppressed
the inflammatory reactions, oxidative damage and plaque burden and
decreased the amount of insoluble amyloid [7]. |
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| To assess a spectrum of learning and memory functions a battery of
tests is needed. These tests are selected to assess behavior and memory.
The present studies were undertaken to assess the neuroprotective
effect of curcumin administrated with Al represented by behavior and
memory tests, confirmed by the histological study of mice’s brains. |
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| Materials and Methods |
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| Chemicals |
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| AlCl3+6H2O were purchased from Merck Chemical Co (Germany). |
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| Curcumin and the other chemicals were purchased from Sigma-
Aldrich Chemical Co. |
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| Animals and treatment |
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| Fifty five healthy adult mice weighting from 18-22 g were obtained
from Pasteur institute Algeria. They were maintained at 24 ± 5°C with
a 12 h light/dark cycle, and have been given a commercial pellet diet
(ONAB Algeria) and fresh drinking water ad libitum. |
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| The mice were randomly divided into four groups; each group
containing seven mice (for each experience: neurotoxicity, Alzheimer’s
model): control group, neurotoxicity and Alzheimer model,
intoxicated/Alzheimer treated groups and the control treated groups.
AlCl3 dissolved in distilled water administrated orally (100 mg/kg) for
the intoxicated/Alzheimer’s model groups, and intoxicated/Alzheimer’s
treated groups, with a D-Galactose IP (200 mg/kg) for the Alzheimer’s
model given for 11 weeks; in parallel of curcumin administration (45
mg orally-200 mg/kg IP) respectively for the intoxicated treated group
and Alzheimer disease animal model. The control treated groups
received the same doses of curcumin (45-200 mg/kg). |
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| Behavioral tests |
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| Functional behavioral assessment is required as part of testing the nervous status (manifest determinations). These guidelines apply
to animals in special tests, including those with locomotors activity,
curiosity (holes test), anxiety/curiosity (maze on +), anxiety (black/
white room) and Persolt test. |
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| The assessment of animal memory using different types of mazes
has been used in neurosciences [8]. Several models have been proposed
recently, mainly trying to evaluate accuracy of choice between the
alternatives presented in the same day of the session, instead of looking
for the accumulated learning through successive days of training. |
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| The Morris water maze is widely used to study spatial memory and
learning. Animals are placed in a pool of water that is colored opaque
with powdered non-toxic tempera paint, where they must swim to a
hidden escape platform. |
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| Histological analysis |
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| Mice from both studies were sacrificed with an overdose of Chloral. |
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| After the step-through avoidance test, the brains of mice were
removed right after sacrifices. Removed brains were then impressed
in 10% neutral buffered formaldehyde for 24 hours. Serial coronal
paraffin sections were cut at 4 μm thickness for hematoxylin and eosin
(H&E) and Congo red staining. |
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| Statistical analysis |
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| The experimental results were reported as mean SEM. Statistical
analysis was performed using SPSS Software. Analysis of variance
(ANOVA) and LSD test were used to compare the experimental groups
with the controls. One-way ANOVA p value using the post hoc Fischer’s
LSD test. P-value<0.05 was considered signifiant. |
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| Results and Discussion |
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| Behavioral tests |
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| Performance of mice in the step through passive avoidance training
and testing show a remarkable difference between the intoxicated
treated/Alzheimer treated and the intoxicated/Alzheimer groups; the
test of maze plus was assessed by the time spending in a covered arm
squares noted as score per time of 20 mn, 5 mn each phases showed high
activity for the intoxicated treated comparatively with the intoxicated
group (Figure 1). |
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|
Figure 1: Result of maze with two arms test raised in first and second steep. |
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| Memory tests |
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| When non-spatial memory test preferably conditional the results
obtained during the three experimental tests show that mice intoxicated
take much longer to reach the food in the arm lit unlike control mice,
treated intoxicated and that put a very short time to get informed on
the arm (Figure 2). |
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Figure 2: Result of non-spatial memory test preferably conditional A: mice
poisoned (I1) by AlCl3 (100 mg/kg VO), poisoned and treated (IT1, IT2) by
curcumin (60mg/kg and 45mg/kg) orally treated (Tr1) by curcumin (45 mg/
kg VO) compared with controls for 11 weeks.
B: mice Alzheimer’s model
(alz) induced by AlCl3 (100 mg/kg Oral)+D-Gal (200 mg/kg IP) Alzheimer
treated (Alz.trt) by curcumin (200 mg/kg) IP, and treated (Tr1) by curcumin
(200 mg/kg) IP for 30 days. |
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| Histological studies |
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| H&E staining shows that there are typical neuropathological
changes in the hippocampus of intoxicated/Alzheimer’s model. In
the control groups the neurons were full and arranged tightly, the
nuclei were light stained. By comparison in the model group mice
the cytoplasm of neuron were shrunken, the nuclei were side moved
and dark stained, neurofibrillary degeneration and neurons loss were
observed in hippocampus. Curcumin administration shows moderated
neuropathological changes. The neurons recovered their characteristic
shape, with prolonged neurofibrillary tangles (Figure 3). Congo red
staining shows the beginning of injuries induced by Aluminium,
characterized by assembling of nuclei resulting from cytoplasm lysis, and a slowly reaction of neurodegenerative mechanism for the
Alzheimer treated model. The control cell wasn’t stained with the
Congo red (Figure 4). |
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Figure 3: Microscopic study of cerebral cortex performed by staining (HE)
in mice poisoned with AlCl3 orally (100 mg/kg) and intoxicated treated mice
(IT) with curcumin for 11 weeks with dose (45 mg/kg) (T) cerebral cortex
of control mice (G×400). (I) cerebral cortex characterized by a decrease in
cell density and neuronal degeneration (G×400), (IT1) shows a decrease of
edema and vacuolation in intoxicated treated (G×400). |
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Figure 4: Microscopic study of nervous tissue staining by (RC) left (G×400)
and right (G×1000); Alzheimer mice (alz) control mice (T), Alzheimer
treated group (alz.trt) and the treated group. (alz) presents a collection
of nuclei resulting from the lysis and cytoplasmic, deposition of amyloid in
homogeneous form, (alz.trt) presents an early gathering of nuclei swelling
but no amyloid deposition, (T) and (Trt) have not taken the Congo red
staining, normal cellular distribution. |
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| Discussion |
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| In the present study, the effects of Al exposure were investigated
to describe the associated behavioral (Figures 1 and 2) and brain
modifications. Neurodegenerative disease is characterized by
progressive pathological changes in the brain (Figures 3 and 4) that
translate into clinical signs of decline in cognitive abilities (memory),
functional abilities, mood, behavior, and finally physical changes. The
pathological changes in the Alzheimer's brain include deterioration
and loss of neurons (nerve cells) leading to brain atrophy [8]. |
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| Dietary antioxidants cooperate with the body enzymes to protect
the brain from free radical damage [9]. Curcumin (of curcuma longa)
is used to improve mental concentration and memory capabilities. It
has free radical scavenging and lipid peroxidation activities [10,11].
Also it is a potent antioxidant with low or no side effects, it increases
antioxidant enzymes, hypoglycemic, antibacterial and antifungal
properties [12,13]. The antioxidant activity of coriander could protect
liver from oxidative damage [14] and from lead induced testis oxidative
damage. |
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| In this investigation the effect of curcumin with over load of
aluminum chloride to mice lead to reduction of neurotoxicity and
Alzheimer's disease appeared as shrunken decreasing of pyramidal
cells, reduced effect of decreasing number of the pyramidal cells. These
brain moderation changes induced by curcumin against aluminum
chloride administration were due to reducing oxidative damage which
contribute to disease pathogenesis and were in accordance with the aim
of this study which is to prove the protective effect of curcumin against aluminium neurotoxicity [15], and also Alzheimer’s disease by means
of curcumin administration with Aluminium chloride [16]. |
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| Mice were poisoned with Aluminium chloride (AlCl3) orally at 100
mg/kg/day in drinking water, another group poisoned with the same
manner but treated with curcumin orally (45 mg/kg/day) for 11weeks.
The animals of the control group received drinking water only during
this period, and the control treated were received curcumin in distilled
water (45 mg/kg). Results showed that curcumin moderated the Al
effect on anxiety/curiosity of treated intoxicated animals (Figure 1).
The same results were shown for the subchronic exposure to AlCl3 in mice Alzheimer’s model increase anxiety in opposite of the results
obtained for the Alzheimer treated group. |
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| Based on these results, it seems likely that enhanced in exploratory
behavior in treated poisoned mice during the whole board test and
locomotor activity may reflect the less anxiety response of an animal
to an unfamiliar environment. The test was based on the adversive
properties of the open field and in which antioxidant treatment -induced
ease of exploratory activity is compared between treated poisoned and
poisoned groups. This model permits simple and quick evaluation of
the anxious behavior and its modification by pharmacological agents.
In our present study forced swimming test showed a significantly
increased immobility time of Al-treated mice. This test is a common
behavioral test for assessing depression in which animals have given up
the hope of escape and depression remains controversial [17], drugs with
antioxidant activity reduce the time during which the animals remain
immobile [18]. These observations were important for the evaluation of
the cognitive tests in since impaired physiological functions or changed
motor performance may conflict with learning and memory tests [19].
Most learning paradigms that require configural associations require a
fully functioning hippocampus; however learning paradigms that can
be solved using only elemental associations can be solved without input
from this structure [20]. The memory task can test the hippocampus in learning and memory. Results of the conditional memory documented
that Al-treated mice showed no impairment in learning and memory
in reference version of the test (Figure 2). |
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| There is no animal model available that can mimic all the cognitive,
behavioral and histopathological abnormalities observed in patient
with Alzheimer disease [20]. |
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| Our histopathological study of chronic exposure shows obvious
difference between the treated poisoned groups and the poisoned, not very normal as a control and control treated, (Figure 3) but can
also explain a deceleration of amyloid reaction, unlike the sub chronic
exposure that wasn’t very clear with H&E staining, which Congo red
staining that showed (Figure 4) the amyloid formation and apoptosis
for the poisoned group (Alzheimer’s model) less important in treated
Alzheimer’s model [21]. |
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| Conclusion |
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| The distribution of aluminum in the tissues, as well as for the
gavage infusion depends on the chemical species used and route of
administration. We can therefore say that the route and duration
of administration, and other interactions result in little damage in
different tissues whose concentration depends on the chemical form. |
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| Turmeric is being studied for the prevention of neurotoxicity
and Alzheimer's, or to slow disease progression. Studies in humans
have been launched, after promising trials in animals, but it’s less
bioavailability and it’s fast eliminated by the liver detoxification
pathways, obliged there administration with piperine exist in Pepper
nigrum, inhibitor of the key enzyme involved in detoxification, which
helps maintain a high rate of curcumin (Curcuma longa) longer |
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| The aim of this research was to evaluate the preventive effect of
curcumin with a fixator of absorption on chronic neurotoxicity of
aluminum, as well as Alzheimer's disease induced (subacute/chronic
sub), through the use of biological models in behavioral, memory, and
histological status of nerves. |
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| References |
| |
- Silva VS, Cordeiro JM, Matos MJ, Oliveira CR, Gonçalves PP (2002) Aluminum accumulation and membrane fluidity alteration in synaptosomes isolated from rat brain cortex following aluminum ingestion: effect of cholestrol. Neurosci Res 44: 181-193.
- Kawahara M, Kato-Negishi M (2011) Link between Aluminum and the Pathogenesis of Alzheimer's Disease: The Integration of the Aluminum and Amyloid Cascade Hypotheses. Int J Alzheimers Dis 8: 276393.
- Rondeau V, Iron A, Letenneur L, Commenges D, Duchêne F, et al. (2006) Analysis of the effect of aluminum in drinking water and transferrin C2 allele on Alzheimer's disease. Eur J Neurol 13: 1022-1025.
- Wu TY, Chen CP, Jinn TR (2011) Traditional Chinese medicines and Alzheimer's disease. Taiwan J Obstet Gynecol 50: 131-135.
- Pan R, Qiu S, Lu DX, Dong J (2008) Curcumin improves learning and memory ability and its neuroprotective mechanism in mice. Chin Med J (Engl) 121: 832-839.
- Sahin Kavakli H, Koca C, Alici O (2011) Antioxidant effects of curcumin in spinal cord injury in rats. Ulus Travma Acil Cerrahi Derg 17: 14-18.
- Turgut G, Kaptanoglu B, Turgut S, Enli Y, Genç O (2004) Effects of chronic aluminum administration on blood and liver iron-related parameters in mice. Yonsei Med J 45: 135-139.
- Exley C (2005) The aluminium-amyloid cascade hypothesis and Alzheimer disease. Subcell Biochem 38: 225-234.
- Xu Y, Ku B, Tie L, Yao H, Jiang W, et al. (2006) Curcumin reverses the effects of chronic stress on behavior, the HPA axis, BDNF expression and phosphorylation of CREB. Brain Res 1122: 56-64.
- Green RJ, Stanton ME (1989) Differential ontogeny of working memory and reference memory in the rat. Behav Neurosci 103: 98-105.
- Ho SC, Liu JH, Wu RY (2003) Establishment of the mimetic aging effect in mice caused by D-galactose. Biogerontology 4: 15-18.
- Tian J, Ishibashi K, Ishibashi K, Reiser K, Grebe R, et al. (2005) Advanced glycation endproduct-induced aging of the retinal pigment epithelium and choroid: a comprehensive transcriptional response. Proc Natl Acad Sci U S A 102: 11846-11851.
- Wei H, Li L, Song Q, Ai H, Chu J, et al. (2005) Behavioural study of the D-galactose induced aging model in C57BL/6J mice. Behav Brain Res 157: 245-251.
- Eskes R, Antonsson B, Osen-Sand A, Montessuit S, Richter C, et al. (1998) Bax-induced cytochrome C release from mitochondria is independent of the permeability transition pore but highly dependent on Mg2+ ions. J Cell Biol 143: 217-224.
- Kumar A, Dogra S, Prakash A (2009) Protective effect of curcumin (Curcuma longa), against aluminium toxicity: Possible behavioral and biochemical alterations in rats. Behav Brain Res 205: 384-390
- Ringman JM, Frautschy SA, Cole GM, Masterman DL, Cummings JL (2005) A potential role of the curry spice curcumin in Alzheimer’s disease. Curr Alzheimer Res 2: 131-136.
- Rebai O, Djebli NE (2008) Chronic Exposure to Aluminum Chloride in Mice: Exploratory Behaviors and Spatial Learning. Adv Biol Res 2: 26-33.
- Gardier AM, Bourin M (2001) Appropriate use of "knockout" mice as models of depression or models of testing the efficacy of antidepressants. Psychopharmacology (Berl) 153: 393-394.
- Borsini F, Meli A (1988) Is the forced swimming test a suitable model for revealing antidepressant activity? Psychopharmacology (Berl) 94: 147-160.
- Yamada K, Nabeshima T (2000) Animal models of Alzheimer’s disease and evaluation of anti-dementia drugs. Pharmacol Ther 88: 93-113.
- Fernandez-Flores A (2010) Comparative study of Congo red fluorescence and immunohistochemistry in cutaneous amyloidosis. Rom J Morphol Embryol 51: 683-686.
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