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Fatty acid as antioxidant

Prevention of Oxidative Stress by Omega-3 Fatty Acids in the Brain:

Neural membranes contain glycerophospholipids, which contain a saturated fatty acid at the sn-1
position of glycerol moiety, whereas the sn-2 position bears a polyunsaturated fatty acid
(arachidonic acid, ARA, 20:4, n-6) or docosahexaenoic acid, DHA (22:6, n-3). ARA belongs to
the omega-6 family whereas DHA falls under the omega-3 family of essential fatty acids,
respectively. Vegetable and fish oils are the most common sources of omega-6 and omega-3
fatty acids, respectively. Fatty acids belonging to omega-6 and omega-3 family not only provide
neural membranes with their physical characteristics, such as acyl chain order and fluidity,
stability, phase behavior, elastic compressibility, ion permeability, fusion, rapid flip-flop and
packing, but also function as signaling molecules by supplying lipid mediators, which are formed
by the oxidation process. An appropriate ratio of ARA to DHA in the brain promotes
development, ameliorates cognitive functions, and provides protection against neurological
diseases by enhancing repairing processes. In addition, ARA and DHA also stimulate gene
expression, boost synaptogenesis, neurogenesis, and induce and prevent oxidative stress, a
process that results from an unbalance between prooxidant and antioxidant systems in the brain.
Neurological oxidative stress is either induced by the failure of cell antioxidant (buffering)
mechanisms or overproduction of reactive oxygen species (ROS), which are oxygen-based
radicals formed in most mammalian cells through the activities of enzymes involved in the
mitochondrial electron transport chain, epoxygenase (EPOX), lipoxygenase (LOX) or
cyclooxygenase (COX), NAD (P) H oxidases or uncoupled nitric oxide synthase (NOS), and
peroxidases, among others. They are normal by-products of healthy cellular metabolic processes
and are known to play physiologically useful roles in cell signaling; for example, as part of the
immunity-oriented “oxidative burst”. Low levels of oxidative stress are needed for cell functions.
The role of oxygen in cell survival is linked to its high redox potential, which makes it an
excellent oxidizing agent capable of accepting electrons easily from reduced substrates. High
levels of oxidative stress are central disruptor of neural cell homeostasis. It is well known that
brain represents only*2 % of the total body mass and yet accounts for more than 25 % glucose
and 20 % of the total consumption of oxygen. In addition to high oxygen utilization, two more
reasons make the brain most susceptible to oxidative damage. First is its modest antioxidant

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defense mechanism and second is the presence of high levels of lipids, which account for 60–65
% of brain dry weight. Thus, the brain is particularly vulnerable to oxidative stress. The major
sources of ROS in brain are uncontrolled ARA cascade, mitochondrial respiratory chain,
xanthine/xanthine oxidase, myeloperoxidase, and activation of NADPH oxidase. Long-term
consumption of Western diet, which is enriched in saturated fats, ARA, cholesterol, and high
simple sugars results in the incorporation of ARA in neural membrane phospholipids
(phosphatidylinositol and phosphatidylcholine). ARA is released and oxidized by phospholipase
A2 and cyclooxygenases-2 (PLA2 and COX-2) respectively. Nonenzymic oxidation of
arachidonic acid results in the generation of high levels of ROS, increased expression of
inflammatory cytokines (TNF-α, IL1β, and IL-6), and elevated levels of n-6 fatty acid-derived
proinflammatory lipid mediators (eicosanoids and platelet-activating factor) throughout the body
including brain. Nonenzymic peroxidation of ARA is a radical-initiated and autocatalytic
process. Free radicals are molecules that contain an unpaired electron in their outer orbit. Free
radicals are associated with many neurochemical activities of cells such as signal transduction,
gene transcription, and regulation of soluble guanylate cyclase activity. Humans are constantly
exposed to free radicals not only generated from the man-made environment, but also from
natural resources such as radon, cosmic radiation, as well as cellular metabolisms (respiratory
burst, enzyme reactions). The most common reported cellular free radicals are hydroxyl (OH ),
superoxide (O2 − ) , and nitric monoxide (NO ) [1, 2]. ARA is highly susceptible to radical-
mediated oxidation due to the abstraction of the bis-allylic methylene hydrogen by other radical
species. The resulting conjugated pentadienyl fatty acid radical (L ) can further react with
oxygen to give a dienyl peroxyl radical (LOO ). This intermediate abstracts a radical from
another lipid generating a lipid hydroperoxide (LOOH) and a secondary radical. The lipid
hydroperoxide (LOOH) undergoes rearrangement and cleavage reactions that result in the
formation of lipoxidation end products. This process generates several products, such as 4-
hydroxynonenal, malondialdehyde, acrolein, isoprostane, isofuran, and isoketal along with
excessive production of ROS. High levels of ROS not only cause more lipid peroxidation and
oxidation of neural membrane protein and DNA, but also produce impairment in normal neural
cell functions leading to neural cell death. Modifications of neural membrane protein and DNA
were originally considered solely as markers of oxidative insult. However, recently the
modifications of proteins and DNA by lipid peroxidation products are recognized as a new

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mechanism of cell signaling with relevance to redox homeostasis, adaptive response, and
inflammatory resolution. Among the neural cells, astrocytes are most resistant to ROS attack
than neurons because they contain higher levels of reduced glutathione (GSH) content than other
neural cells. During scavenging of ROS, the reduced form of GSH is converted into oxidized
form of glutathione (GSSG). Thus, high levels of ROS generation promote disruption of redox
signaling in neural cells. The main impact of high levels of n-6 fatty acid consumption in
Western diet is the generation of high levels of superoxide in the mitochondria. Superoxide is a
relatively unstable intermediate and in large part is converted to hydrogen peroxide in the
mitochondria by superoxide dismutase. The newly formed hydrogen peroxide undergoes a
Haber–Weiss or Fenton reaction, generating a highly reactive hydroxyl radical, which can
oxidize mitochondrial proteins, DNA, and lipids and amplify the effects of the superoxide-
initiated oxidative stress. Molecular mechanisms associated with nuclear and mitochondrial
DNA (mtDNA) are not fully understood. However, it is suggested that promoter regions with
high guanine–cytosine contents in DNA are specifically vulnerable to oxidative damage.
Oxidative damage to promoter regions of DNA may cause changes in conformation leading to
loss of DNA affinity for transcription factors. Furthermore, oxidative damage to mitochondrial
DNA (mtDNA) may result in downregulation of genes related to respiratory chain leading to
impairment in energy metabolism. In addition, telomeres, regions of repetitive nucleotide
sequences at each end of a chromosome, are highly susceptible to oxidative stress because of
their high content of guanines. Increase in intracellular ROS levels is associated with
acceleration in the rate of telomere shortening. Oxidative stress-mediated progressive shortening
of telomeres leads to senescence, apoptotic cell death, or the oncogenic transformation of
somatic cells in various tissues. As stated above, ARA is released by the action of PLA2 on
neural membrane phospholipids. It is oxidized by COXs and LOXs into proinflammatory
eicosanoids (prostaglandins) and leukotrienes) or nonenzymically oxidized into lipid mediators
of arachidonic acid metabolism along with production of ROS. The generation of high ROS
activates redox-sensitive transcription factor that results in numerous downstream effects,
including the increased expression of proinflammatory cytokines leading to further increase in
ROS production. Production of ROS is also promoted by RAGE receptors, which are activated
by advanced glycation products (AGEs). AGEs are generated by an irreversible reaction through
the nonenzymatic, long-term glycosylation of proteins. They are strongly resistant to proteolytic

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processes and induce protein cross-linking. They inhibit the physiological functions of many
proteins and receptors promoting the transformation of soluble proteins into insoluble proteins
deposits, as well as activating the microglia through specific ligands for cell surface receptors.
The complex interplay between inflammatory mediators and markers for oxidative stress caused
by the long-term consumption of Western diet has been proposed to regulate the progression of
chronic neurodegeneration in neurodegenerative diseases. In addition, nitric oxide (NO)
synthesis occurs in mitochondria from the breakdown of arginine into citrulline by a family of
NADPH-dependent enzymes called mitochondrial nitric oxide synthases (mtNOS). Once
formed, NO inhibits respiration by binding to heme groups in the proteins of the electron
transport chain, including cytochrome c oxidase. Thus, NO is a known inhibitor of the
respiratory chain. NO competes directly with O2 at complex IV, reversibly retarding the
formation of this complex and inducing ROS generation. During high oxidative stress, NO reacts
rapidly with excess superoxide to form peroxynitrite, a powerful oxidizing and nitrating agent,
which irreversibly inhibits multiple complexes of the respiratory chain, as well as dismutase
enzymes. These processes not only lead to elevation in oxidative stress and increase in
mitochondrial inner membrane potential, but also induce alterations in calcium homeostasis.
Collective evidence suggests that oxidative stress and inflammation caused by distinct
biochemical cascades are processes, which are closely intertwined and generally function in
parallel, particularly in the brain, an organ especially prone to oxidative stress.

Enzymic and Nonenzymic Lipid Mediators of ARA-Derived Metabolism and

Their Antioxidant and Anti-inflammatory Effects

Stimulation of glutamatergic, serotonergic, cholinergic, or dopaminergic receptors by their

agonists in the brain results in the activation of Ca2+-dependent PLA2 and release of arachidonic
acid. The released ARA is oxidized by COXs, LOXs, and EPOXs resulting in the formation of
prostaglandins (PGs), leukotriene (LTs), lipoxins (LXs), and thromboxanes (TXs), as well as
hydroxyl-eicosatetraenoic acid, epoxyeicosatetraenoic acids, and dihydroxyeicosatrienoic acids.
Most arachidonic acid-derived lipid mediators produce prooxidant, prothrombotic,
proaggregatory, and proinflammatory effects. However, arachidonic acid-derived LXs produce
anti-inflammatory effects. PGs, LTs, and TXs interact with their receptors and produce potent
effects on neuroinflammation, vasodilation, vasoconstriction, apoptosis, and immune responses.

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The expression of COX-2 is increased during acute inflammation and the ingestion of aspirin
leads to acetylation of COX-2, which blocks PG formation. Acetylated COX-2 is catalytically
active. It transforms ARA into 15(R)-HETE rather than PGs. 15(R)-HETE is utilized by 5-LOX
for transforming into 15(R)-LXA4. Aspirin-triggered 15-epi-LXs are *twofold more potent than
15(S)-LXs. ROS also attack on lipid hydro peroxides. This attack results in the synthesis of
isoprostanes (IsoPs) via β-cleavage of the peroxyl acid and subsequent molecular rearrangement.
IsoP contains D-, E-, and F-ringed structures similar to cyclooxygenase-generated
prostaglandins, except that their hydrocarbon chains are in the cis position in relation to the
pentane ring as opposed to the Trans position observed in prostaglandins. The formation of IsoP
is used as a “gold standard” to quantify cumulative oxidative stress in neurological diseases.
Neurochemical activities of IsoPs are poorly understood. However, they may form covalent
adducts with cysteine residues in proteins through Michael-type addition reactions.

Prevention of Oxidative Stress and Neuroinflammation by EPA and DHA

It is well known that in Western diet, the ratio between ARA and DHA is about 20:1. In contrast,
Paleolithic diet on which humans have lived and survived thousands of years contained the ratio
between ARA and DHA of 1:1. Due to the presence of high levels of ARA, long-term
consumption of Western diet results in the generation of high levels of ARA-derived enzymic
(PGs, LTs, and TXs) and nonenzymic (4-HNE, IsoP, MDA, etc.) lipid mediators, which promote
neuroinflammation and oxidative stress, along with apoptotic cell death. As stated above,
oxidative stress and neuroinflammation are closely intertwined processes that generally function
in parallel, particularly in the brain, an organ especially prone to both oxidative stress and
neuroinflammation. These processes are also closely associated with the pathogenesis of
neurotraumatic, neurodegenerative, and neuropsychiatric diseases. In addition, consumption of
Western diet induces obesity, which correlates with reduction in focal gray matter volume and
enlargement in white matter in the frontal lobe, altering learning, memory, and executive
function in humans and cognitive deficits in a rodent model. Furthermore, long-term
consumption of Western diet also decreases levels of BDNF in the hippocampus leading to
impairment in neurogenesis. Consumption of the Western diet also mediates significant decrease
in Nrf2 DNA-binding activity, reduces Nrf2 responsive pathway proteins (heme oxygenase-1
and NAD (P)H dehydrogenase, Quinone 1), and decreases the expression of Nrf2 protein

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expression as depressive symptoms. Collective evidence suggests that long-term consumption of

Western diet increases the chances of brain damage through oxidative processes along with
changes in neural cell homeostasis leading to metabolic abnormalities, behavioral disturbances,
and motor and cognitive impairments. Increase in fish consumption or inclusion of fish oil in the
diet not only decreases levels of ARA-derived enzymic and nonenzymic lipid mediators, but also
generates DHA- and EPA-derived lipid mediators (D-series Rvs, E-series Rvs, NPD1, and
MaRs). These omega-3 fatty acid-derived lipid mediators produce antioxidant and anti-
inflammatory effects and retard the onset and pathogenesis of neurotraumatic,
neurodegenerative, and neuropsychiatric diseases. It is also proposed that consumption of
omega-3 fatty acid-containing diet counteracts signals that respond to oxidative stress and set in
motion brain repair and neuroplasticity responses nand RvE2) and DHA-derived resolvins
(RvD1 and RvD2) produce potent antioxidants, anti-inflammatory, and pro-resolution effects.
They inhibit oxidative stress, retard excessive inflammation, and promote resolution by
enhancing clearance of apoptotic cells and debris from inflamed brain tissue. These properties
result in the beneficial effects of EPA and DHA in human health and neurotraumatic and
neurodegenerative diseases.