Module 4 Reading Materials (Supplementary)

Free radicals and antioxidants in human disease

Free radicals and endogenous antioxidant defense systems

Under physiological conditions, oxygen-derived free radicals or reactive oxygen species

(ROS) such as superoxide, hydrogen peroxide, and hydroxyl radicals are metabolic
byproducts, arising mainly from the mitochondrion (the powerhouse of the cell), in all
aerobic organisms including humans. By definition, a free radical is any atom, molecule
or compound that possesses at least one unpaired electron. Due to the high tendency
of the unpaired electron to acquire a partner electron, free radicals are highly reactive
and readily extract an electron from a “victim” molecule forming a chemically stable
product. In addition to energy metabolism, oxygen- and nitrogen-centered free radicals
are involved in other biochemical reactions, such as those involved in cytochrome
P450-mediated arachidonic acid and xenobiotic metabolism.

To cope with the excessive accumulation of potentially damaging free radicals, aerobic
organisms have evolved a host of antioxidant defenses that consist of both enzymatic
and non-enzymatic components. While antioxidant enzymes, including catalase,
superoxide dismutase, selenium-dependent glutathione peroxidase, glutathione
transferases, and glutathione reductase, can decompose or detoxify free radicals and
other oxidant species by virtue of their catalytic potential, enzymatic antioxidant
components also include proteins that are responsible for repairing oxidative damage in
cellular structures and macromolecules. Non-enzymatic antioxidants are small
molecules, such as ascorbic acid (vitamin C) and α-tocopherol (vitamin E), which can
scavenge free radicals by direct chemical interaction. Besides serving as free radical
scavengers, some naturally-occurring non-enzymatic antioxidants such as uric acid can
bind transition metal ions, thereby rendering them catalytically inactive in free radical-

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generating reactions. Likewise, glutathione, a tripeptide free radical scavenger, can also
participate in enzyme-catalyzed reactions involved in the detoxification of free radicals
and other reactive oxidant species.

Oxidative stress and human disease

Under physiological conditions, the human body has barely adequate antioxidant
defenses to counteract the normal rate of production of free radicals. However, there is
not a large excess of antioxidant defense capacity. As such, it is easy to tip the balance
in favor of free radicals, resulting in a condition of “oxidative stress”. Under conditions of
oxidative stress, the uncontrolled action of free radicals can cause oxidative damage to
carbohydrates, lipids, proteins, and nucleic acids. Most cells can tolerate a minor
degree of oxidative stress because they have repair systems that recognize, remove
and replace damaged molecules. In addition, cells can adaptively increase antioxidant
defenses in response to stress. However, the prolonged exposure of cells to increased
oxidative stress, as occurs in a number of disease states when an increase in free
radical production and/or impairment in antioxidant defense is known to occur, can
result in sustained oxidative damage with the disruption of cellular structural and
functional integrity of cells, tissues and organs. In this regard, ncontrolled free radical-
mediated processes have been implicated in more than 60 diseases affecting humans,
particularly disorders such as cancer, cardiovascular diseases, neurodegenerative
diseases and immuno-incompetence which increase both in incidence and severity with
increasing age.

Scientific rationale of antioxidant supplementation

As the “Free Radical Theory of Aging” has developed, antioxidants have become a
household name in many countries, and they have assumed an ever-increasing
awareness by the lay public. Both endogenous vitamin as well as plant-derived
(phytochemical) antioxidants are widely used as antioxidant supplements. The
promotion of antioxidant supplementation in preventive health often relies on data from

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epidemiological studies, which attempt to relate the occurrence of certain diseases in a
given population to antioxidant intake. Two early prospective epidemiological studies,
based on dietary questionnaires from female nurses and male health professionals,
suggested that the use of vitamin E supplements lowered the risk of coronary heart
disease in these two populations of females and males aged 34-59 and 40-75,
respectively. There is also strong epidemiological evidence that a diet of fruit and
vegetables can prevent a range of human cancers. It is presumed that dietary
supplementation with antioxidants such as vitamin C and E, β-carotene, etc., could
protect against the toxic or mutagenic effects of free radicals, which are generated
either endogenously in the body or by exogenous chemicals present in food, water or
the environment.

Although the lay public received such epidemiological findings with great enthusiasm, it
should be noted that questionnaires about diet are not a very reliable way of measuring
intake of antioxidants. Moreover, the main problem with epidemiology is that correlation
does not imply causation. Only intervention can prove the cause/consequence
relationship, and then only after very careful analysis of the results. To overcome this
criticism a clinical trial dealing with vitamin supplementation was undertaken in a rural
county of Linxian, China, which has high death rates from cancer of the esophagus and
stomach. A mixture of selenium, β-carotene, and vitamin E produced a significant
decrease in deaths from stomach cancer after 5 years, whereas supplementation with a
vitamin C-containing mixture did not. The effect of antioxidant supplementation on
cancer mortality in this poorly nourished population was convincing, but it is uncertain
which component(s) of the supplement were actually responsible for the effect.
Nevertheless, the results of this study do suggest the potential benefit of antioxidant
nutrients from dietary sources in the prevention of cancer. It is particularly intriguing in
this regard that many compounds occurring naturally in red wine, grapes, tea, coffee,
chocolate, soy, grains, onions, citric fruits, broccoli and tomato also possess antioxidant
activity and therefore may possess anti-cancer activity.

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However, results showing detrimental effects have also been obtained in some
antioxidant intervention trials. One Finnish study attracted a great deal of attention for
showing a deleterious effect of β-carotene supplementation. The results indicated that
vitamin E supplementation had no significant effect on the incidence of lung cancer in
males who were heavy cigarette smokers. Remarkably 3 years into the study, subjects
receiving β-carotene showed a significantly greater incidence of lung cancer than
controls. While the mechanism involved in the deleterious effect of β-carotene
supplementation remains unclear, the lesson is that antioxidants from non-dietary
sources at relatively high doses must be supplemented cautiously. In this example,
vitamin E (dl-α-tocopherol) supplementation was found to displace γ-tocopherol from
lipid membranes, thereby compromising the cell’s ability to trap the mutagenic
electrophiles effectively. Thus, supplementation with dl-α-tocopherol alone may
eventually lead to increases in the risk of cancers.

Oxidative processes – while frequently damaging - are also important in the metabolism
of endogenous compounds and also in cell signaling, etc. Excessive levels of
antioxidants may be detrimental by disrupting these essential processes in vivo.