SITI ANNISA DEVI TRUSDA

BIOCHEMISTRY DEPARTMENT

FREE RADICALS AND

ANTIOXIDANTS
Oxygen molecule
 Ways of filling the outer shell:
 Gaining or losing electrons
 Sharing its electron
 Maximum stability
 Normally, bonds don’t split in away that
leaves a molecule with an odd, unpaired
electron
 Weak bond splitfree radicals formed
 very unstable, react quickly with other
compound
 Free radicals are highly reactive due to the
presence of unpaired electron(s)
 Free radicals involving oxygen = reactive
oxygen species (ROS)
 Oxygen centered free radicals contain two
unpaired electrons in the outer shell
Set off chain reaction
 Free radicals can affect tissues, lipids,
proteins and DNA
 Cell damagedisease/aging, increased over
time
Functions of Free Radicals

 Some free radicals arise normally during

metabolism
 The body’s immune system’s cells purposefully
create them to neutralize viruses and bacteria
 Normally, the body can handle free radicals,
but if antioxidant are unavailable, or if the
free-radical production becomes excessive,
damage can occur
 free radical damage accumulate with age
Types : Endogenous and Exogenous
Types of endogenous free radicals

 Most important: Reactive oxygen species:

 Oxygen : triplet (3O2), singlet (O2), hydroxyl radical
(OH), nitric oxide (NO), hypochlorous acid (HOCl),
hydrogen peroxide (H2O2), and the superoxide radical
(O2-)
 Carbon centered free radicals (from an oxidizing radical
on an organic molecule)
 Hydrogen centered radicals (from attack of the H atom)
 Sulfur centered radical (from oxidation of
glutathionethiyl radical)
Exogenous free radicals

 Drugs
 Antibiotics that depend on bound metals for
activity : nitrofurantoin
 Antineoplastic agent : bleomycin, anthracyclines,
adriamycin, methotrexate
 Penicillamine, phenylbutazone, sulphasalazine
may inactivate protease and deplete vit Clipid
peroxidation
Exogenous free radicals

 Radiation
 Radiotherapy
 Electromagnetic radiation (X rays, gamma rays)
and particulate radiation (electrons, photons,
neutrons, alpha and beta particles)primary
radicals by transferring their energy to cellular
components such as water
secondary reactions with dissolved oxygen or
with cellular solutes
Exogenous free radicals

 Tobacco smoking
 Aldehydes, epoxides, peroxides, etc
 Relatively stable radicals in the tar phase
 Elevated amounts of neutrophils
↑ concentration of free radicals
Exogenous free radicals

 Inorganic particles (via inhalation)

 Asbestos  pulmonary fibrosis
 Silica phagocytosed by pulmonary macrophage
ruptureproteolytic enzyme and chemotactic
mediatorsROS ↑
Exogenous free radicals

 Gases:
 Ozone (not free radical but a very powerful
oxidising agent). Contain 2 unpaired electrons,
degrades to OHfree radicals
 Ozone can generate lipid peroxidation
 Others
 Fever, excess glucocorticoid th/, hyperthyroidism
 Other chemical air pollutans, pestisides, solvents,
anaesthetics, exhaust fume
Free Radicals and Disease
Antioxidants, Nature and Chemistry

 The role of antioxidants is to detoxify reactive

oxygen intermediate (ROI) in the body
 A substance that when present in low
concentrations relative to the oxidizable
substrate significantly delays or reduces
oxidation of the substrate
 Combat oxidation
 Antioxidant “sacrifice” itself to become
oxidized
 One antioxidant for one free radicals the
amount of antioxidant is not unlimited
 Important to replenish antioxidant
Antioxidant System

 Endogenous antioxidant systems:

 1. Superoxide dismutase (SOD), which
catalyses the conversion of O2- to H2O2 and
H2O-
 2. Catalase, which then converts H2O2 to
H2O and O2
 3. Glutathione peroxidase, which reduces
H2O2 to H2O
Endogenous Antioxidant

 The glutathione redox cycle is a central

mechanism for reduction of intracellular
hydroperoxides
 Source and Nature:
 Tetrameric protein, that confers catalytic activity
 Glutathione peroxidase reduces H2O2 to H2O by
oxidizing glutathione
 Re reduction of the oxidized form of glutathione
(GSSG) is then catalyzed by glutathione reductase
Exogenous antioxidant
 1. Lipid soluble vitamins, vit E and Vit A or
provit A
 2. Water soluble vitamin C
 3. Glutathione (GSH) a tripeptide molecule

 Enzymatic and non enzymatic antioxidant

systems are intimately linked to one another
Classification of major antioxidants
Antioxidant Vitamins

 Vitamin E, more appropriately described as an

antioxidantdoes not act as a co-factor for
enzymatic reactions
 Deficiency vit E slow developing symptoms
within decades e.g: atherosclerosis
 Overt symptoms of vit E deficiency found in fat
malabsorption, premature infants and patient on
total parenteral nutrition
 Funcion: prevent lipid peroxidation of the cell
membrane
 Tocopherol-OH can transfer hydrogen atom
with a single electron to free radical  removing
free radical before interacts with cell
membrane/protein/ lipid peroxidation
 Tocopherol-OH + free radical = tocopherol-O
(free radical)
 Vit C + tocopherol –O = semi-dehydro-ascorbat +
tocopherol-OH
 aggressive ROI eliminated, weak ROI
(dehydroascorbat) is formed
 tocopherol-OH generated
 Vit E stimulates immune response lower
incidence of infection when vit E levels are high
 Vit E inhibit cancer initiationenhance
immuno-competence
 Vit E inhibits conversion of nitrites in smoked,
pickled and cured foods to nitrosamines in
stomach
 Alpha tocopherol reduce Fe3+ to Fe 2+ (pro
oxidant)
Beta Carotene (Pro vit A)

Source and Nature

 Carotenoids are pigmented micronutrients
present in fruits and vegetables
 >600 carotenoid found in food : α carotene,β
carotene, lycopene, crocetin, canthaxantin,
zeaxanthin
 β carotene : 2 molecul vit A joined together
(retinol)
 Dietary beta carotene is converted to retinol in
intestine
MoA
 Quenching singlet oxygen,
 Scavenge free radical
 Protect cell membrane lipids
 Limited ability because autoxidation
Vitamin C (Ascorbic acid)

Source and Nature

 Citrus fruits, potatoes, tomatoes, green leafy
veg
 MoA
 Water soluble, chain breaking antioxidant:
scavenges free radicals and oxygen molecules
 Prevents formation of carcinogens from precursor
compounds
 Vit C more potent than alpha tocopherol in
inhibiting the oxidation of LDL
 Regeneration of membrane bound oxidized
vit E.
 Vit C supplementation leads to increased
plasma and tissue levels of vitamin E
Vitamin C and E correlation
Other antioxidants

 Glutathione (GSH): defence against dietary

xenobiotic
 CoQ10 (ubiquinone)
 Albumin and plasma protein (ceruloplasmin
and transferrin)
 Melatonin
 Uric Acid in small amount
 Drugs : xanthin oxidase and statin
Antioxidant Cost and Benefit
 1. Antioxidants are reducing agent that can also act as
pro-oxidant
 Vit C reduces H2O2, but will also reduce metal ionsfree
radicals
 Paradoxical

 2. Free radicals induce an endogenous response which

protects against exogenous radicals (and possibly other
toxic compounds)

 3. Another fact:
 Antioxidant supplements have no clear effect on the risk of
chronic diseases

Hormesis

 Generally favourable biological responses to low

exposures to toxins and other stressors
 Physiologically: exercise, acute stress and uric
acid, all of which in small amounts increase
lifespan, immunity and health, but in larger
amounts are harmful
 Activate the repair mechanism of the body
 Free radicals may also help by causing apoptosis
of damaged cellsreducing aging and cancer
causing cells
IMPORTANT POINTS

 Diet plays an important role in reducing

oxidative damage
 The role of dietary antioxidants is not limited
to its antioxidant chemicals, minerals and
vitamins alone, but to an as yet unknown
combination of all these
 Suppelmentation pills of individual
antioxidants have not been shown to prolong
life or improve health