Antioxidants
• Compounds that prevent oxidative damage to cellular
components by decreasing oxidative stress
• Neutralise the reactive species formed during the various
metabolic processes.
• These compounds may be synthesized in the body or obtained
from the diet.
• Even when present in low concentration relative to an
oxidizable substrate, significantly delay or reduce the oxidation
of the substrate.
• Capable of protecting biomolecules from damaging oxidation
reactions by neutralising free radicals and other reactive species.
MECHANISM OF ACTION:
a) Chain breaking mechanism in which the anti-
oxidant donates an electron to the free radical
b) Removal of reactive oxygen and nitrogen species by
quenching chain-initiating catalysts.
The antioxidants which stop the chain reaction are
termed primary antioxidants.
The antioxidants which eliminate the intiation or
reduce the oxygen level and prevent the formation
of reactive species are termed secondary
antioxidants.
CLASSIFICATION
on the basis of their composition and physio-chemical
properties.
1) ENZYMES:
Superoxide dismutase (SOD), Catalase, glutathione
peroxidase (GPx) etc.
SOD catalyses the transformation of superoxide radical
into H₂O₂.
O₂•⁻+ O₂ •⁻ + 2H⁺ SOD H₂O₂ + O₂
Catalase dismutates H₂O₂ to water and molecular oxygen
2H₂O₂ CATALASE 2H₂O + O₂
GPx reduces lipid peroxides (ROOH) to a stable and non-
toxic molecule, hydroxyl fatty acid (ROH).
2GSH + ROOH GPx GSSG + H₂O + ROH
2) VITAMINS:
– Vit. E (tocopherol) is the major antioxidant which breaks
the chain of lipid peroxidation and scavenges lipid
peroxides, O₂•⁻ and OH•.
– Vit A (β-CAROTENE) binds to transition metals and
scavenges OH•, O₂•⁻ and peroxy radicals.
– Asorbic acid (Vit. C) directly scavenges O₂•⁻ , OH• and
H₂O₂. It also contributes to regeneration of tocopherol.
3) MISCELLANEOUS:
– Other molecules which act as major antioxidants are
thiols (eg. Glutathione), albumin, ceruloplasmin,
transferrin, haptoglobin, bilurubin, uric acid and some
polyphenols.
a) THIOLS:
– Thiols are compounds containing a sulphydryl (-SH) GROUP
Attached to a C-atom
– Thiols exist in vivo in three forms
(1) The free form, R-SH
(2) As homodisulphides, R-S-S-R OR
(3) As heterodisulphide, R₁-S-S-R₂ .
– Some sulphur containing antioxidants are cysteine,
methionine, taurine, lipoic acid and N-acetyl cysteine
– Other thiols are methanethiol, ethanethiol, coenzyme A
and 2-mercaptoindole.
– Thiols interact by redox and disulphide exchange and they
comprise a dynamic system called the redox thiol status.
GLUTATHIONE :
– Glutathione (γ-glutamyl-cystinyl-glycine) is a ubiquitous
sulphydryl antioxidant.
– It is soluble and found mainly in the cell cytosol and other
aqueous phases of the living system.
– It exists in two forms. The antioxidant reduced glutathione
(GSH) and the oxidized form, glutathione disulphide
(GSSG).
– GSH plays an important rule in elimination of lipid
peroxides (ROOH).
– The reaction is catalysed by glutathione peroxidase.
2GSH + ROOH GPx GSSG + H₂O + ROH
– The disulphide is reduced to gsh by glutathione
reductase.
GSSG + NADPH + H⁺ GR 2GSH + NADP
– GSH depletion can trigger suicide of the cell by a process
called apoptosis.
FUNCTIONS:
– The cellular functions linked to the reducing power of
GSH are cytoprotection ; protein and DNA synthesis; cell
growth and division; reductive metabolism; leukotriene
synthesis; regulation of sulphydryl enzymes and
conjugation of xenobiotics.
B) ALBUMIN:
– Albumin is a small globular protein with a molecular weight
of 66.3 kDa.
– It is the most important protein found in the plasma and
accounts for approximately one half of the plasma protein
mass.
– It is highly soluble in water due to its high net negative
charge at physiological pH .
– Albumin can provide antioxidant protection by functioning
as a serum peroxidase in the presence of reduced
glutathione which is an intracellular antioxidant
– Albumin is a metal binding protein and acts as a preventive
antioxidant.
– It can also exert its antioxidant activity through its -SH
group.
Role of oxidative stress
in diseases
CANCER
Cancer is a multistage process defined by at least
three stages: initiation, promotion, and
progression.
Oxidative stress interacts with all three stages of
this process.
During the initiation stage, ROS may cause DNA
damage by introducing gene mutations and
structural alterations of the DNA.
• In the promotion stage, ROS can contribute to
abnormal gene expression, blockage of cell- to
cell communication, and modification of
second messenger systems, thus resulting in
an increase of cell proliferation or a decrease
in apoptosis of the initiated cell population.
• Finally, oxidative stress may also participate in
the progression stage of the cancer process by
adding further DNA alterations to the initiated
cell population.
ATHEROSCLEROSIS
• Oxidative stress plays an important role in the
pathogenesis of atherosclerosis especially by
promoting the oxidative modification of low-
density lipoprotein (LDL). Oxidation of LDL is
one of the earliest events in atherogenesis
and NADPH oxidase has been demonstrated
to be critically involved in this process by
acting either directly or indirectly as a
precursor of ROS.
• ROS also act as early inducers of autophagy.
Autophagy is a pathway for bulk degradation
that helps in eliminating long-lived proteins,
macromolecular aggregates, and damaged
intracellular organelles.
• AUTOPHAGY of oxidized LDL by macrophages
leads to their transformation to foam cells
• Aggregation of foam cells further leads to
plaque formation
DIABETIC COMPLICATIONS
• Oxidative stress plays a pivotal role in the development
of diabetes complications, both microvascular and
cardiovascular.
• The metabolic abnormalities of diabetes cause
mitochondrial superoxide overproduction in endothelial
cells of both large and small vessels, as well as in the
myocardium.
• This increased superoxide production causes increased
formation of AGEs (advanced glycation end products)
which are involved in the pathogenesis of complications.
• It also directly inactivates two critical anti-
atherosclerotic enzymes, endothelial nitric
oxide synthase and prostacyclin synthase.
• Increased intracellular reactive oxygen species
(ROS) cause defective angiogenesis in
response to ischemia and activate a number
of proinflammatory pathways.
• Overexpression of superoxide dismutase in
transgenic diabetic mice prevents diabetic
retinopathy, nephropathy, and
cardiomyopathy.
