Polyphenol Ellagic Acid-Targeting To Brain: A Hidden
Treasure
Sidharth
Mehan, Ramandeep Kaur, Shaba Parveen, Deepa Khanna, Sanjeev Kalra
Sidharth
Mehan, Ramandeep Kaur, Shaba Parveen, Deepa Khanna, Sanjeev Kalra, Rajendra Institute Of Technology & Sciences,
Sirsa-125055, Haryana, India
Correspondence to: Sidharth Mehan, Associate
Professor, Department of Pharmacology, Rajendra Institute of Technology &
Sciences, 4th Mile Stone, Hissar Road,Sirsa-125055, Haryana, India
Email: sidh.mehan@gmail.com
Telephone: +918059889909
Received: March 11,
2015
Revised: May 16, 2015
Accepted: May 18, 2015
Published online: September 1, 2015
ABSTRACT
Alzheimer¡¯s
disease is a severe neurodegenerative disorder that gradually results in loss
of memory and impairment of cognitive functions in the elderly.Thedeposition of
amyloid plaques is the primary event that leads to an oxidative and
inflammatory reactions, neurofibrillary tangle formation, and ultimately
neuronal death. The most prominent losses occur with cholinergic neurons that
plays an important role in the formation of memory and cognitive
functions.Insulin receptor numbers also altered, thusglucosemetabolism is
defected in AD. Polyphenol ellagic acid (EA) possesses anti-oxidant,
anti-inflammatory, anti-carcinogenic, anti-diabetic and cardioprotection
activities.Activities of EA that are linked with protection of neuronal
abnormalities like anti-depressant, anti-anxiety, anti-nociception. Ellagic
acid inhibits ¦Â-secretase enzymatic activities, thus prevents the main
pathologic hallmark of AD. There are varieties of herbal extracts and
phytochemicals formulations extensively used to provide symptomatic relief to
AD patients. Together these results our hypothesis that EA may prevent the
neuronal dysfunctions as well as further work is also required to examine the
pharmacological properties of EA in the protection of specially
neurodegenerative disorder like AD. Here, we discuss how we can use the
knowledge to improve treatment strategies of EA in AD and to explore the
various signaling pathways involved in the progression of neuronal death.
© 2015 ACT. All
rights reserved.
Key words: Alzheimer¡¯s disease; Ellagic acid; Neuro-inflammation;
Amyloid plaques
Mehan S, Kaur R,
Parveen S, Khanna D, Kalra S. Polyphenol Ellagic Acid-Targeting To Brain: A
Hidden Treasure. International Journal of Neurology
Research 2015; 1(3):
141-152 Available from: URL: http://www.ghrnet.org/index.php/ijnr/article/view/1107
INTRODUCTION
Alzheimer¡¯s disease is a severe neurodegenerativedisease that gradually
results in loss of memory andimpairment of cognitive functions in the elderly[1].
According to Alzheimer¡¯s Association, Alzheimer¡¯s disease (AD) is affecting an
estimated 5.2 million Americans in 2014, including 5.0 million over age 65[2].
AD include extracellular amyloid deposition in senile plaques and
intra-neuronal neurofibrillary tangles of hyperphosphorylated
microtubule-associated tau protein[3-6]. The extracellular
deposition of amyloid plaques is a major event that progress an inflammatory
reaction, intracellular neurofibrillary tangle formation, and ultimately cause
neuronal death[7,8]. The neuronal loss produces insufficiencies in
several neurotransmitter systems[8]. However, the most prominent
losses occur with cholinergic neurons in the basal forebrain, which project
into the hippocampus and neocortex, the brain structures that play an important
role in memory and cognitive function[9,10]. The loss of cholinergic
neurons results in up to 90% reduction in the activity of an enzyme, choline
acetyltranferase (ChAT), needed for the synthesis of the neurotransmitter
acetylcholine[11]. Treatment strategies have focused on boosting
acetylcholine, by the development of cholinesterase inhibitors.
Acetylcholinesterase (AChE) inhibitors work by reversibly bindingto the choline
binding subsite of acetylcholinesterease, thus preventing degradation of
acetylcholine[12]. Themechanisms of neuronal cell loss in AD have
not yet beenfully elucidated, but increased oxidative stressandinflammationare
considered important mediators of neuronal damage in AD[13]. The
brains of individuals with AD had increasedlevels of lipid peroxidation
products such as 4-hydroxynonenalor 2-propenal, and enhanced lipid
peroxidationwas also detected in the cerebrospinal fluid and plasma
ofindividuals with AD[14,15]. In addition, oxidativedamage to
membrane proteins, cellular mitochondrial and nuclear DNA isalso an important
event in AD pathology[1].
The first
neurotransmitter defect discovered in AD involved acetylcholine[16]
(Francis et al, 1999). As cholinergic function isrequiredfor short-term
memory, the cholinergic deficitin AD was also believed to be responsiblefor
much of the short-term memory deficit[16,17]. Clinical drug trials
in patients with AD have focused on drugs that augmentlevels of acetylcholine
in the brain to compensatefor the loss ofcholinergic function[18].
These drugs have included acetylcholine precursors, muscarinic agonists,
nicotinic agonists, andacetylcholinesterase inhibitors[9]. Herbal
drugs are alsopre-clinically as well as clinically tested and shown to be very
effective as Galantamine[19,20], Ginkgo biloba[21,22],
Bacopa monnieri (brahmi)[23,24], Celastrus paniculatus[25,26],
Curcumin[27-29], coffee[30-32] and Lavender essential oil[33].
Many naturally
occurring compounds have been proposed as potential therapies to slow or
prevent the progression of AD, mostly by acting as antioxidants[34],
but also with some direct anti-amyloid actions[35]. Recent studies
have suggested the positive effects of dietary antioxidants as an aid in
potentially reducing somatic cell and neuronal damage by free radicals[36].
The beneficial health effects of plant-derived products have been largely
attributed to polyphenolic compounds, as well as vitamins, minerals and dietary
fibers[37]. Polyphenols occur as products of the plant secondary
metabolism in response to biotic and abiotic stress[38,39]. Polyphenols
are the main compounds of green tea and there are a class of polyphenolic
flavonoids known as catechins (the most abundant component) which comprise of (−)-epigallocatechin-3-gallate
(EGCG), (−)-epigallocatechin (EGC), (−)-epicatechin-3-gallate (ECG) and (−)-epicatechin
(EC) respectively. Green tea polyphenols may play a protective effect on
cultured rat primary prefrontal cortical neurons against A¦Â-induced
cytotoxicity and AKT is involved in the green tea polyphenols-induced
protective effects[40]. Epigallocatechin-3-gallate protects neuronal
cells by disrupting amyloid-¦Â fibril formation; bioactive polyphenols which
disrupt fibril aggregation may confer additional neuroprotection when compared
against antioxidant activity alone[35]. Grape seed polyphenol
extract (GSPE) has a significant potential for therapeutic development by
neutralizing phosphoepitopes and disrupting fibrillary conformation leading to
disintegration of paired helical filaments (PHFs)[41]. LMN diet rich
in polyphenols, dry fruits and cocoa, was able to decrease behavioral
deterioration caused by aging and Tg2576 genotype and to delay the A¦Â plaque
formation. These results corroborate the increasing importance of polyphenols
as human dietary supplements in amelioration of the cognitive impairment during
aging and neurological disorders such as AD[42]. At present, there
is mounting evidence that alterations in dietary intake could be beneficial for
preventing and treating oxidative disorders and neurodegenerative illnesses
such as AD, which could offer an alternative or supplementary therapy.
Natural
polyphenols have pleiotropic activity including antioxidant properties.
Particularly, natural polyphenols significantly attenuated cognitive
impairments and amyloid-beta burden. Natural polyphenolic compounds exhibit
their antioxidant effect by quenching free radical species and/or promoting
endogenous antioxidant capacity. Thus, the antioxidant properties certainly
contribute to their neuroprotective effects[43].
Ellagic acid
(EA), is a powerful non-flavonoid polyphenol compound with various
pharmacological actions and said to be a degraded product of certain plant
tannins[44] (Table 1). Ester bonds hydrolysis of ellagitannins with
acids or bases produce hexa-hydroxydiphenic acid (HHDP), which rearranges
spontaneously to produce ellagic acid[45]. EA is present in fruit,
food and beverages but berries are potential dietary source of quercetin or
ellagic acid. High concentration of ellagic acid is found in cloudberry
(60mg/100g), blackberry (42.4mg/100g), strawberry (19.8/100g) red raspberry
(17.9mg/100g), cranberry (12mg/100g), boysenberry (30mg/kg), marion blackberry
(32mg/kg), evergreen blackberry (21mg/kg)[45-47], while, blueberry
(0.9mg/100g), walnut (590µg/g) and pecans (330µg/g) have low concentration of
ellagic acid[48,49]. It is also found in seeds of red raspberry
(8.7mg/kg) and black raspberry (6.7 mg/kg)[45]. Juice of red
muscadine grapes and fresh arils of pomegranate also contains ellagic acid in
concentration of 10.2 mg/100g and 15.3 mg/L respectively[48,50].
Various extracts are also rich in ellagic acid as hot water extract of guava
contains 36.68 mg/g and muscadine grape cultivas extract contains 49.7 mg/kg
while, ethanolic extract of longan seeds have 1.6mg/g of ellagic acid in
concentration[45,51]. Ellagic acid is present either in free form or
as conjugate of sulphate ester, glucuronide, and glutathione[45,52].
Ellagic acid being a weak acid gets ionized at physiological pH, form poorly
soluble complexes with calcium and magnesium ions in the intestine and bind to
intestinal epithelium, thereby it is poorly absorbed[52,53]. Cerda
et al., (2005) showed the presence of metabolites of ellagic acid in bile and
urine suggesting that its absorption take place in stomach[55].
Bioavailability of this phenolic components from plants is low due to its
occurrence in glycosidic form[56] but bioavailability studies
conducted by Aiyer et al (2008) revealed that ellagic acid availability
is more as compare to chlorogenic acid which, may be due to its absorption in
upper part of the gut, in the stomach and small intestine[57]. After
being absorbed, EA is metabolized by phase II enzymes, such as glucuronosyl
transferases and sulfotransferases[53]. Ellagic acid is released
from ellagitannins in jejunum, and is metabolize to yield urolithin D.
Chemically urolithin D is tetrahydroxydibenzopyran-6-one, more lipophilic and
well absorbed than EA. Urolithin D is further metabolize to urolithin C
(trihydroxydibenzopyran-6-one), urolithin A (dihydroxydibenzopyran-6-one) and
urolithin B (monohydroxydibenzopyran-6-one) respectively. The removal of
hydroxyl groups from metabolites increases the lipophilicity and thereby their
absorption as compare to ellagic acid[52]. Thus the presence of
urolithin plays an important role in absorption. Further, the presence of
urolithin A, C, and D metabolites in bile show an active entro-hepatic
circulation, this cause low clearance of metabolites. Only urolithin A shows
its presence in feces[52]. The half life of ellagic acid is 8.4¡À1.8
hrs with 50% plasma protein binding[58]. Solubility studies
conducted by Bala et al., (2005) demonstrated that Ellagic acid was soluble in
various pharmaceutically acceptable solvents including triethanolamine,
polyethylene glycol 400 and N-methyl pyrrollidone (NMP). Ellagic acid showed
poor solublity in water (9.7µg/mL) and due to its acidic nature it is soluble
in phosphate buffer while, highly soluble in methanol[59]. Ellagic
acid is soluble at physiological pH and it remain insoluble at pH less than 7,
while at pH more than 7, ellagic acid chelates with divalent cations such as Zn2+,
Ca2+, Fe2+, Cd2+, Cu2+ which,
contributes in its anti-carcinogenic property[60].
Ellagic acid
plays an essential role in explaining the sensory properties of fruits, food,
beverages which exhibit this phytoconstituent. The major contribution of
berries as antioxidant is due to ellagic acid[46]; antioxidant
effects of pomegranate juice was due to ellagic acid[50] likewise,
Gauva leaf extract exhibit antioxidant potential due to ellagic acid[51].
Favarin et al, (2013)[61] and Corbett et al, (2010)[62]
evaluateanti-inflammatory potential of ellagic acid in animal models. Several in
vivo and in vitro studies mention ellagic acid as anti-proliferative[60,63,64].
Further studies designed to found ellagic acid¡¯s cardiopreotective role prove
that it is a novel therapeutic strategy to manage myocardial infarction and
atherosclerosis[65,66] (Figure 1). The present review mainly
summarize the signaling mechanisms involved in the pathogenesis of Alzheimer¡¯s
and find out the better way to overcome that destructive neuronal changes in AD
brain via ellagic acid treatment.
ELLAGIC ACID AND AMYLOID BETA
(A¦¢) PEPTIDE
Neuropathological hallmarks of AD are accumulation of extracellular
¡®¡®senile plaques¡± containing amyloid ¦Â-peptide (A¦Â) fibrils and intracellular
¡®¡®neurofibrillary tangles¡± containing hyper-phosphorylated tau protein[67].
Neurologicalevents that contribute to AD progression include amyloid-beta (A¦Â)
aggregation, acetylcholine depletion, and oxidative stress[68].
Amyloid beta-protein (A¦Â) is the major component of senile plaques and
cerebrovascular amyloid deposits in individuals with Alzheimer¡¯s disease[1].
A¦Â fibril formation is a multi-step process that is preceded by oligomerization
and aggregation of monomeric A¦Â, and it involves conformational change of the
peptide from alpha-helical to beta-pleated sheet structure[69].
Recent evidences suggest that solubleoligomers of A¦Â in the brain are
neurotoxic and a major risk factor for the onset and progression of cognitive
deterioration in AD[70]. A¦Â is known to increase free radical
production in neuronal cells[1].
Possible
benefit of EAtreatment was demonstrated in Sprague-Dawley rats, infused with A¦Â
followed by control solution had significantly impaired spatial working and
reference memory, where the EA treatments restored to normal condition. This
study not only demonstrated detrimental effects of a single A¦Â mixed peptide
infusion, but also some beneficial effects of multiple EA treatments in
reducing these A¦Â-induced deficits. Further, A¦Â-treatment decreased working and
reference memory capabilities while EA treatment restored those capabilities.
Also consistent with the hypotheses, the measure of ROS activity (MDA content
via the TBARS assay) in the hippocampus was significantly increased in animals
treated with A¦Â and only control aCSF (Artificial cerebrospinal fluid) solution
whenA¦Â-treated animals were infused with Ellagic Acid, they showed MDA levels
that were significantly lower than the A¦Â+aCSF group and comparable to the aCSF
animals[68].
Pomegranate
juice (PJ) consisted of polyphenols (phenolic acids and flavonoids). Phenolic
acids included 115 ppm ellagic acid and 5ppm gallic acid. Flavonoids included
1880 ppm hydrolysable tannins (e.g., gallotannins, ellagitannins, punicalagin)
and 369 ppm anthocyanins and their glycosides (e.g., cyanidin, delphinidin,
pelargonidin). Tg2576/APPsw mice, commonly used as a model of A¦Â deposition and
associated AD-like pathology, were given pomegranate juice in their drinking
water. PJ improved behavioral performance of APPsw mice in the water maze. PJ
decreased A¦Â and amyloid levels in the hippocampus. When their learning
behavior was assessed PJ-treated mice exhibited improvements on cued and
spatial learning tasks as well as faster overall swim speeds. Additionally,
plaque load (both A¦Â and fibrillar A¦Â/amyloid) and soluble A¦Â42 were
significantly reduced in the hippocampus[71].
In-vitro
activity of human recombinant ¦Â-secretase (BACE1) was studied using a
fluorogenic substrate based on the cleavage site for the enzyme in the Swedish
mutation of amyloid precursor protein. Amyloid plaques are formed by overproduction
and subsequent aggregation of the 40-42 amino acid peptide,¦Â-amyloid (A¦Â). ¦Â-amyloid
is produced by cleavage of the membrane-anchored amyloid precursor protein
(APP) by the ¦Â-site APP-cleaving enzyme (BACE1), also known as ¦Â-secretase,
which is subsequently cleaved by ¦Ã-secretase to form either A¦Â 40 or A¦Â 42. The
longer form of A¦Â is implicated as causative in AD through association with
early onset AD. In vitro studies have shown that A¦Â42 aggregates into
fibrils faster than A¦Â40 and it is considered to be more toxic than A¦Â40.
Non-competitive inhibitors of BACE1 were identified from natural products,
including polyphenolics: ellagic acid and punicalagin from pomegranate husk.
Pomegranate husk inhibited BACE1 with an EC50 of 1.19 µg.mL-1,which
could be partly explained by the inhibitory activity of ellagic acid with an EC50
value of 0.85µg/mL (2.8µM). Ellagic acid is heterocyclic polyphenolics with
extensive peripheral substitution by hydroxyl groups, which may account for
inhibitory activity. Thus, Ellagic acid is a strong inhibitor[72].
EA prevented A¦Â
neurotoxicity by promoting A¦Â aggregation into fibrils with significant
oligomer loss. Thus EA could significantly reduce A¦Â42-induced neurotoxicity
toward SH-SY5Y cells[67].
Walnuts
(Juglans regia L.) are an excellent source of ¦Á-linolenic acid (plant-based
omega-3 fatty acid) and have a high content of antioxidants such as flavonoids,
phenolic acid (ellagic acid), melatonin, gamma tocopherol and selenium.
Walnutextract can counteract A¦Â-induced oxidative stress and associated cell
death[1]. EA inhibit ¦Â-secretase (BACE1) in vitro, thus inhibiting A¦Â-fibrillation[73].
Therefore, ellagic acid can be a potential therapeutic treatment that inhibits
amyloid beta-protein (A¦Â), which is the major component of senile plaques and
amyloid deposits in AD patients.
ELLAGIC ACID AND
NEURO-OXIDATION
During respiration, mitochondria produce semi-reduced oxygen species,
superoxide and hydrogen peroxide that cause the oxidative stress[74].
Brain is susceptible to oxidative damage because of various reasons such as
ease of peroxidation of brain membrane as it is enrich in peroxidizable fatty
acids, high consumption of oxygen and glucose, and areas enriched in ferrous
ascorbate/pro-oxidant in comparison to other body parts and anti oxidant
capacity is lower than oxidant possibilities such as catalase activity, which
is 10-20% of liver and heart[75]. H2O2 is
generated by almost all sources of oxidative stress. DNA is damaged by H2O2
via fenton reaction in presence of oxygen and transition metal ions[76,77].
During neurodegenerative diseases, the various oxidative reactions involved
that leads to neuronal cell death[78]. Some resources such as
vitamins, lipoic acid, antioxidant enzymes and redox sensitive protein
transcriptional factors,help to compensate oxidative stress in neuronal cells[79-83].
Polyphenols,
the nutritional antioxidants can modulate this defence system[78].
Ellagic acid prevents oxidationin liver,heart, kidney testicular and
spermatozoal damages associated with oxidation[84-89].
Terminalia
Chebula contain polyphenols such as quercetin, gallic acid and ellagic
acids[90]. T. Chebula extract has protective effectsagainst
in-vitro experiment where oxygen-glucose deprivation followed by reoxygenation
(OGD-R) ischemia and hydrogen peroxide (H2O2) induced
cell death on PC12 cells. T. chebula extract showed the
diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity and reduced H2O2-induced
MDA production where DPPH radical scavenging activity of T. chebula is
mostly related to the hydroxyl group in ellagic acid[91]. Moreover,
Ellagitannins produces hexa-hydroxydiphenic acid (HHDP) on ester bonds
hydrolysis with acids or bases, which rearranges spontaneously to produce
Ellagic acid[45]. Ellagitannins showed remarkable neuroprotective
activity via inhibition on the H2O2-induced oxidative
damage in NG108-15 hybridoma cell line[92].
Diabetic
neuropathy and encephalopathy are common life threatening complications of diabetes
mellitus[93]. Ellagic acid exhibits neuro-protective effects against
oxidative damage in streptozotocin-induced diabetic rats. Ellagic acid has the
ability to reduce the oxidative damage to the diabetic rat sciatic and brain
tissues. Malonaldehyde (MDA), total oxidant status (TOS), oxidative stress
index (OSI), and nitric oxide (NO) levels in brain and sciatic tissues were
found to be significantly reduced in the ellagic acid-treated diabetic group
compared to the untreated diabetic group in these tissues[94].
Ellagic acid possesses non-enzymatic antioxidant activity such as scavenging
free radicals, and enzymatic antioxidant activitysuch as increasing protein
level of antioxidant enzymesSuperoxide Dismutase (SOD), Catalase
(CAT),Glutathione and Glutathione Peroxidase(GSH-PX)[86].
The rat
pheochromocytoma line PC12 provides a useful model system for investigating
neuronal cell injury.Tert-Butyl hydroperoxide(t-BHP) may act through formation
of peroxynitrate (ONOO-) and hydrogen peroxide (H2O2)
through hydroxyl radicals, their effects in PC12 cells are cytotoxicity via
lipid peroxidation and free radical generation[95]. EA has strong
free radical scavenging and protective activities towards oxidative damage
through t-BHP and FeSO4 in PC12 cell culture of neuronal origin.
FeSO4, suspected to produce superoxide radicals, induced comparable
changes in viability and GSH content and enhanced lipid peroxidation as
measured by MDA production and ROS production. ROS formation and enhanced lipid
peroxidation are not the direct cause of cell death in PC12 cells, while loss
of GSH much better correlates with cytotoxicity, where EA significantly
restored the antioxidant defense modulation. EA can also chelate metal ions and
prevent iron- and copper- catalysed formation of ROS[96].
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a persistent environmental
contaminant that has been found to produce adverse effects in brain. TCDD
caused significant increases in superoxide anion production in cerebral cortex
and hippocampus[97]. EA provided protection against TCDD-induced
superoxide anion production in brain tissues. Ellagic acidresults in
significant reduction in the production of superoxide anion in cerebral cortex
and hippocampus when administered simultaneously with TCDD, as compared with
corresponding regions of the TCDD-treated animals. EA administered in
combination with TCDD, the compounds resulted in significant reductions in
TBARS production in cerebral cortex and hippocampus, as compared with the corresponding
regions in the TCDD-treated group. EA result in significant reductions in
TCDD-induced DNA single-strand breaks in cerebral cortex and hippocampus[98].
On the basis of above review, Ellagic acid can be a capable therapeutic
treatment in AD patients, where various oxidative reactions involved that leads
to neuronal cell death.
ELLAGIC ACID AND
NEUROINFLAMMATION
Alzheimer disease (AD) brain is characterized by extracellular plaques
of amyloid ¦Â (A¦Â) peptide with reactive microglia[99]. Cognitive impairment
in neurodegenerative diseases such as Alzheimer¡¯s disease may be partly due to
long-term exposure and increased susceptibility to inflammatory insults[100].
The association between inflammation and AD manifests itself through (1) the
presence of activated microglia surrounding senile plaques and (2) higher
levels of inflammatory mediators in the brains of AD patients versus
age-matched controls[5,101,102].
Oxidative
damage due to the ROS/RNS play the vital role to initiate a wide range of
intracellular stress signaling processes that culminate in excessive cytokine
such as inducible COX-2 and iNOS and adhesion molecule as intracellular
adhesion molecule (ICAM)-1 and E- andP-selectins upregulation[91,103].
Neuroinflammatory processes have been shown to have a fundamental role in the
pathogenesis of neurodegenerative disorders like Alzheimer¡¯s disease (AD)[104].
Ellagicacid
inhibits upregulation of COX-2 and reduces IL-6 and increase IL-10 levels in
0.1N HCl induced acute lung injury in mice modelvia its anti-inflammatory
mechanism[61]. Ellagic acid decrease TNF-¦Á, inhibit COX-2 & iNOS
and block the activation of NF-¦ÊB pathway in trinitrobenzene sulfonic acid
induced chronic colonic inflammation in rats[105]. Ellagic acid also
reduce the expressions of NF-¦ÊB, COX-2, iNOS, TNF-¦Á and IL-6 in
1,2-Dimethylhydrazine-induced rat colon carcinogenesis[106].
Moreover ellagic acid inhibits TNF-¦Á induced endothelial activation and
expression of ICAM-1 and VCAM-1 adhesion molecules in Human aorta endothelial
cells (HAEC)[107].
The activation
of the primary microglia is one of the major reasons of cellular inflammation
in ischemic condition. Terminalia chebula extract reduced LPS induced cell
death and inhibited the production of NO by BV2 cellsin Lipopolysaccharide
activated microglia cells (BV2 cell) and reduce cellular inflammation[91].
Pomegranates
extract feeding improved behavioral performance in APP/PS1 mice, which
correlated with decreased microgliosis, nuclear factor of activated T-cell c2
(NFATc2) activity, and brain TNF-¦Á concentration. Thus pomegranate extract
suppressed nuclear factor of activated T-cell c2 (NFATc2) expression,
attenuated brain cytokine concentrations and decreased A¦Â plaque load in the
amyloid precursor protein/presenilin 1 (APP/PS1) mice and decreased
microgliosis in the APP/PS1 mice.Pomegranate extract component, ellagic acid
attenuated microglial proinflammatory cytokine release in-vitro as ability to
inhibit NFAT activity and cytokine secretion in-vitrocell culture of C57BL/6
mouse brains microglia. Possible mechanism for these may as ellagic acid, is
absorbed and able to enter the brain to inhibit microglial activation via
attenuation of, particularly, NFAT activity or ellagic acid can be directly
taken up into cells or may be these large polyphenol molecules may also act
extra-cellularly to exert their anti-inflammatory action[99].
Blueberry
poly-phenols, such as ellagic acid reduced activation of the inflammatory
marker (OX-6), expression of the inflammatory cytokines IL-1¦Â and TNF-¦Á, the
transcription factor NF-B and increased expression in the neuroprotective trophic factor IGF-1
in centrally administered kainic acid inflammatory rat model where as ellagic
acid significantly reduced the impairment in their spatial learning and memory
abilities[48,100]. Shukitt-Hale et al, (2008) suggested that
possible mechanism of action of the blueberry polyphenols like ellagic acid
could be reduction of the deleterious effects of an inflammatory stimulus by
altering the expression of genes associated with inflammation, protecting the brain
and/or reducing its susceptibility to insults[100].
ELLAGIC ACID AND
ACETYLCHOLINESTERASE INHIBITION
Cholinergic dysfunction in neurodegenerative diseases can be the result
of reduction in Ach synthesis due to reduced choline acetyltransferase (ChAT)
or choline uptake, cholinergic neuronal and axonal abnormalities, and
degeneration of cholinergic neurons[108,109]. Acetyl cholinesterase
inhibitors have been extensively used for the symptomatic treatment of
Alzheimer¡¯s disease[110-112].
Walnut extract
and EA, one of its constituents, have been shown to inhibit the action of two
sites on AChE that are involved in A¦Â aggregation and acetylcholine
catabolism[68]. In vitro studies found that IC50 value of AchE inhibitory
activity of ellagic acid was 13.79 ¦Ìg/mL(45.63¦ÌM)[113]. EC25
value of Ellagic acid, from Cochlospermum angolensis Welw extracts was found to
be 2362.6 mg/ml (hydromethanolic extract) and 2235.5mg/mL (boiling water
extract), for AChE inhibitory assays[114]. Moreover by using brain
tissue showed a marked decrease in brain AChE activity in the presence of EA[68,114].
These results show the therapeutic efficacy of EA in AD patients.
ELLAGIC ACID AND INSULIN
AD causes alteration in insulin receptor numbers[115].
Recent studies suggested that glucose metabolism is defected during AD[116-118].
GLUT4 translocation from an intracellular pool to the cell surface is the
primary factor responsible for insulin induced glucose uptake in adipocytes and
skeletal muscles. This process can be studied in vitro by employing a chimeric
GLUT4 construct with an epitope at an extracellular domain so that the extent
of the epitope externalization directly matches with the amount of GLUT4
present on the plasma membrane[119]. Due to presence of ellagic acid
(EA) as an active component present in the methanol extract prepared from the
leaves of Terminalia Arjuna stimulate glucose transport in 3T3-L1
adipocytes and C2C12 myotubes.
AMPK
activation is known to stimulate glucose transport in adipocytes and muscles in
an insulin independent manner. EA also stimulated GLUT4 translocation occurs in
an AMPK dependent manner and involves extracellular signal-regulated kinase 1/2
(ERK1/2) and atypical protein kinase C ¦Æ/¦Ë (¦ÁPKC ¦Æ/¦Ë) activation[120].
Protein phosphorylation studies with EA suggest that its stimulatory effect on
GLUT4 translocation is achieved through stimulating AMPK ERK- ¦ÁPKC pathway.
Glucose transport stimulated by EA was inhibited by treatment with AMPK
inhibitor Compound C and MEK inhibitor PD98059. Application of compound C also
inhibits ERK1/2 phosphorylation to a similar extent, suggesting that AMPK lies
upstream to ERK in the EA signaling pathway[121].
Resistin, a
peptidesignaling molecule, is secreted by adipocytes and induced during
adipogenesis (the differentiation of preadipocytes to adipocytes). Resistin
neutralization enhance insulin-stimulated glucose uptake in adipocytes.
Resistin is thus a hormone that potentially links obesity to diabetes[122].
Therefore, reducing circulating resistin levels would be very beneficial for
ameliorating obesity-induced insulin resistance. The consumption of pomegranate
juice and ellagic acid, a main component of pomegranate, reduced serum resistin
levels in ovariectomized mice, the animal model having elevated serum resistin
levels[123]. Type 2 diabetes is potentially preventable by
suppressing elevated serum resistin levels[124,125].
In a study,
pomegranate fruit extract (PFE) is fed to ovariectomized mice as an animal
model with high resistin levels to investigate the effects of pomegranate on
resistin in vivo. Moreover, investigation of mechanism of the action of PFE
usingdifferentiated murine 3T3-L1 adipocytes revealed PFE, particularly EA,
significantly suppress resistin secretion by 3T3-L1 adipocytes without altering
the mRNA expression of resistin. Therefore, EA is a unique component derived
from food material that can regulate resistin by a novel mechanism[123].
Ellagic acid also present in aqueous
Jamun seed (Syzigium Cumini)extract (JSEt) as well as in alcoholic JSEt[126].
Oral administration of an aqueous Jamun seed extract (JSEt) caused a
significant decrease in lipids, thiobarbituric acid reactive substances (TBARS)
and an increase in catalase and superoxide dismutase in the brain of alloxan
induced diabetic rats[127]. Furthermore, EA can be a
promisingtherapeutic diet in AD patients where insulin resistance occurs, due
to imbalance of glucose transportation for other insulin dependent activation.
ELLAGIC ACID AND DEPRESSION
Depression is a common, debilitating, life-threatening illness with a
high incidence. Numerous antidepressant compounds presumably act via different
mechanisms involving the serotonergic, noradrenergic and dopaminergic systems[128].
The
serotonergic (5-HT4) receptor may play a role in memory and learning and 5-HT4
receptor activation has been suggested to modulate acetylcholine release and to
reduce amyloid-¦Â (A¦Â) accumulation[129]. GABA-mediated inhibitory
transmission can be strongly modulated by 5-HT: such modulation, in parallel
with 5-HT-glutamate interactions, has been observed in structures involved in
learning and memory, sensory processing, nociception and motor control[130].
In vitro
antidepressant potential of ellagic acid was found in MAO-A inhibition assay[114].
Further, ellagic acid produce anti-immobility effect in anti-depressant models
mediated through monoaminergic system (serotonergic and noradrenergic systems)[131].
Pretreatment with prazosin (an alpha-adrenoceptor antagonist) and p-CPA
(serotonin synthesis inhibitor) reverse the anti depressive effect of ellagic
acid where ellagic acid interacts with ¦Á-adrenoceptors and serotonergic
receptors to produce anti depressive effect[128].
Emblica
officinalis (EO) fruits mainly contain tannins and vitamin C like
substances in abundance and their chemical constituents include gallic acid,
ellagic acid, emblicanin A, emblicanin B, punigluconin and flavanoids[132].
The aqueous extract of the EO fruits contain 30.0% tannins and 10.0% Gallic
acid[133].
The
development of immobility when rodents were suspended by their tail during TST
(tail suspension test) and placed in an inescapable cylinder of water during
FST (forced swim test) reflects the cessation of their persistent
escape-directed behavior. EO reliably decreases the duration of immobility in
animals during these tests. This decrease in duration of immobility is
considered to have a good predictive value of ellagic acid in the evaluation of
potential antidepressant agents[133].
The ethanolic
extract of stem barks of Lafoensia pacari (PEtExt), which contains Ellagic acid
as major compoundalso possesses antidepressant-like effect[134,135].
Fruits of T.
Bellirica contain about 17% tannin and sitoterol, gallic acid ellagic acid,
ethyl gallate, galloyl glucose and chebulagic acid. The effect of aqueous and
ethanolic extracts of Terminalia bellirica was tested on depression in mice
using FST and TST. Both aqueous and ethanolic extract reversed reserpine
induced extension of immobility period of mice in FST and TST and elicited a
significant antidepressant like effect in mice by interaction with adrenergic,
dopaminergic and serotonirgic systems[136].
In addition to
cholinergic deficits, deficiencies in central GABAergic neural transmission may
also play a critical role in some of the clinical manifestations of AD[137].
Some studies evident for the dysregulation of genes related to the
glutamatergic and GABAergic transmitter systems in depression individuals. As
the ratio of excitatory-inhibitory neurotransmitter levels disturb, cytotoxic
damage to neurons and glia occurs. Significantly reduced numbers of neuroglia
is reported in major depressive disorder[138]. GABA concentrations
are lower in cerebrospinal fluid and plasma in unipolar depressed patients[139].
In a study ellagic acid also produces anti anxiety-like effect which was
antagonized by a non-competitive GABAA receptor antagonist and a benzodiazepine
site antagonist which shows modulation of GABAergic system by EA[140].
Receptorfor
advanced glycation endproducts (RAGE), is a tentative cell membrane receptor
for A¦Â, binds to A¦Â with high affinity[141]. GABA increases the
formation of soluble RAGE and decreases the levels of full-length RAGE,
therefore lowering A¦Â uptakeby RAGE[142,143]. These findings suggest
that GABA receptor modulation can be a potential treatment for AD. Moreover,
ellagic acid can modulate deficiencies in central GABAergic neural
transmission.
ELLAGIC ACID AND NOCICEPTION
Ellagicacid shows central and peripheral anti-nociceptive
effect.Possible mechanisms of anti-nociception activity of ellagic acid are
involvement of opioidergic system and L-arginine¨CNO¨C cGMP¨CATP sensitive K+ channels
pathwayin tail flick test and in writhing test Central nervous system may be
the primary site for EA to exert its analgesic effect and also the activation
of opioid receptors and/or an increment of endogenous opioids might be involved
in the antinociceptive effect of EA[144]. Ellagic acid has also
antinociceptive properties via cyclooxygenase (COX) inhibition. Ellagic acid
with ketorolac significantly shows antinociceptive activity in inflammatory
pain induced by carrageenan[145]. Moreover, EA can be a nascent
therapeutic herbal where excessive cytokine such as inducible COX-2 is up
regulated as in neuroinflammation in AD patients.
FUTURE PROSPECTIVE OF ELLAGIC
ACID IN AD BRAIN
The deposition of amyloid plaques is the primary event of AD that further
starts the other neuronal dysfunctions like inflammatory reaction,
neurofibrillary tangle formation, and ultimately neuronal death[7,8].
The neuronal loss produces insufficiencies in several neurotransmitter systems[8].
Amyloid beta-protein (A¦Â) is the major component of senile plaques and
cerebrovascular amyloid deposits in individuals with Alzheimer¡¯s disease[1].
During neurodegenerative diseases, the various oxidative reactions involved
that leads to neuronal cell death[78]. The presence of activated microglia
surrounding senile plaques and higher levels of inflammatory mediators are
manifestations of AD[5,102]. Ellagic acid, is a polyphenol, inhibit
the amyloid deposition and effective tool as an anti-oxidantand
anti-inflammatory. However, the most prominent losses occur with cholinergic
neurons in the basal forebrain, which play an important role in memory and
cognitive function[9,10] (Table 2) (Figure 2). Ellagic acid also
plays its role as it inhibits the acetyl cholinesterase enzyme thus it can minimize
the effect of deficiency of acetylcholine. Furthermore, EA can be a promising
therapeutic diet in AD patients where insulin resistance occurs, due to
imbalance of glucose transportation and other insulin dependent activation.
Because there are no effective neuroprotective therapies that delay the
progression of the AD, symptomatic treatment remains the cornerstone of medical
management. Need of today¡¯s is diets or drugs that modify age-related
degenerative disease and provide a strong symptomatic relief to AD patients.
There is major requirement to determine therapeutic potential for EA in cases
of AD with suitable biochemical markers. Drug and methodological manipulations
should be explored in future investigations.
CONFLICT OF INTERESTS
The authors have no conflicts of interest to declare.
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Peer reviewer:
Xiaohui Zhang, Professor, State Key Laboratory of Cognitive Neuroscience &
Learning, IDG/McGovern Institute for Brain Research, Beijing Normal University,
Beijing 100875, China.
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