Free Radicals & Antioxidants: Gandham. Rajeev
Free Radicals & Antioxidants: Gandham. Rajeev
Free Radicals & Antioxidants: Gandham. Rajeev
Gandham. Rajeev
Email:[email protected]
A free radical is a molecule or molecular
fragment that contains one or more unpaired
electrons in its outer orbital.
Represented by a superscript dot, (R•).
Oxidation reactions ensure that molecular
oxygen is completely reduced to water.
The products of partial reduction of oxygen
are highly reactive, called Reactive oxygen
species or ROS
Free Radicals
Superoxide anion radical (O2 -•)
Hydroperoxyl radical (HOO•)
Hydrogen peroxide (H2O2)
Hydroxyl radical (OH•)
Lipid peroxide radical (ROO•)
Singlet oxygen ( 1O2)
Nitric oxide (NO•)
Peroxy nitrite (ONOO-•).
H2O2 & singlet oxygen are not free radicals
Because of their extreme reactivity, they are
included in the group of ROS.
Characteristics of ROS:
Extreme reactivity.
Short life span.
Generation of new ROS by chain reaction.
Damage to various tissues.
Generation of Free Radicals
Cellular Metabolism
During phagocytosis
Photolysis of O2 by light
Cigarette smoke
LH + OH- L- + H2O
Propagation phase:
Under aerobic conditions, the fatty acid
radical (L-) takes up oxygen to form peroxy
radical (LOO-).
The latter, in turn, can remove H-atom from
another PUFA (LH) to form lipid
hydroperoxide (LOOH).
LH + OH- L- + H2O
LOO- + LH LOOH + L-
The hydroperoxides are capable of further
stimulating lipid peroxidation as they can
form alkoxy (LO-) & peroxyl (LOO-) radicals.
Termination:
Lipid peroxidation proceeds as a chain
reaction until the PUFA gets oxidized.
Malondialdehyde (MDA)
The products of lipid peroxidation are unstable e.g.
carbonyls, esters, alkanes, alkenes, 2-alkenal ,2 ,4-
alkadienal, MDA.
Malondialdehyde (-CHO-CH2-CHO-) is a marker for
the assessment of lipid peroxidation.
MDA & other aldehydes react with thiobarbituric
acid & produce red-coloured products - thiobarbituric
acid reactive substances (TBARS) which can be
measured colorimetrically.
Generation of ROS by macrophages
During the course of phagocytosis,
inflammatory cells, particularly the
macrophages produce superoxide, by NADPH
oxidase.
This O2 radical gets converted to H2O2 & then
to hypochlorous acid (HCIO).
Superoxide radical along with hypochlorous
ions brings about bactericidal action.
The beneficial affects of the free radicals
generated by the body.
A large amount of O2 is consumed by
macrophages during their bactericidal
function, a phenomenon referred to as
respiratory burst.
Harmful effects of free radicals
Proteins: Free radicals cause oxidation of
sulfhydryl groups & modification of certain
amino acids (e.g. methionine, cysteine,
histidine, tryptophan).
ROS may damage proteins by
fragmentation, cross-linking & aggregation.
Lipids: PUFA are susceptible to damage by
free radicals.
Carbohydrates:
Oxidation of monosaccharides (e .g. glucose)
can produce H2O2 & oxoaldehyde.
Glycation increases the susceptibility of
proteins to the attack by free radicals. E.g. DM.
Nucleic acids:
Free radicals may cause DNA strand breaks,
fragmentation of bases & deoxyribose.
Clinical significance
Chronic Inflammation
Chronic inflammatory diseases such as
rheumatoid arthritis are self-perpetuated by
the free radicals released by neutrophils.
ROS induced tissue damage appears to be
involved in pathogenesis of chronic
ulcerative colitis, chronic glomerulonephritis,
Cardiovascular diseases (CHD):
Oxidized low density lipoproteins (LDL),
formed by the action of free radicals,
promote atherosclerosis & CHD.
Cancer:
Free radicals can damage DNA & cause
mutagenicity and cytotoxicity & play a key
role in carcinogenesis.
Respiratory diseases:
Direct exposure of lungs to 100% oxygen for a
long period is known to destroy endothelium
& cause lung edema.
This is mediated by free radicals.
ROS are also responsible for adult
respiratory distress syndrome (ARDS), a
disorder characterized by pulmonary edema.
Diabetes:
Destruction of islets of pancreas due to the
accumulation of free radicals is one of the
causes for the pathogenesis of IDDM.
Cataract:
Increased exposure to oxidative stress
contributes to cataract formation.
Male infertility: Free radicals reduce sperm
motility & viability & male infertility.
Aging process:
Reactive oxygen metabolites (ROM) play a
pivotal role in the degenerative brain
disorders such as Parkinsonism, Alzheimer's
dementia and multiple sclerosis.
Cumulative effects of free radical injury
cause gradual deterioration in aging process.
Antioxidants
Antioxidants in relation to lipid peroxidation:
Preventive antioxidants that will block the
initial production of free radicals e.g.
catalase, glutathione peroxidase.
Chain breaking antioxidants that inhibit the
propagative phase of lipid peroxidation e.g.
Superoxide dismutase, vitamin E, uric acid.
According to location:
Plasma antioxidants: e.g. β-carotene,
ascorbic acid, bilirubin, uric acid,
ceruloplasmin, transferrin.
Cell membrane antioxidants: e.g. α-
tocopherol.
Intracellular antioxidants: e.g. SOD,
catalase, glutathione peroxidase.
According to nature & action:
Enzymatic antioxidants:
E.g. SOD, catalase, glutathione peroxidase &
glutathione reductase.
Non-enzymatic antioxidants:
Nutrient: carotenoids, α-tocopherol, selenium &
vitamin C.
Metabolic: glutathione, ceruloplasmin, albumin,
bilirubin, transferrin, ferritin, uric acid.
Antioxidant enzymes
Superoxide dismutase:
It converts superoxide (O2) to hydrogen
peroxide & O2.
This is the first line of defense to protect cells
from the injurious effects of superoxide
Catalase:
H2O2 is metabolised by catalase.
Glutathione peroxidase:
It detoxifies H2O2 to H2O, while reduced
glutathione (G-SH) is converted to oxidized
glutathione (GS-SG).
The reduced glutathione can be regenerated
by glutathione reductase utilizing NADPH.
HMP shunt is the major source of NADPH.
References