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Brooke Duncan

Honors Option FSHN 350-001

Fall 2014

The terms oxidative stress, antioxidants, and free radicals have become

informal chemical terms that relate to the environmental, physical, and chemical

strain placed on our bodies. According to Harvard School of Public Health,

antioxidants have become increasingly popular since the 1990s when there was

surge of research in place to determine if there was biological benefits and illness

prevention when foods with increased amounts of antioxidants were used to impede

biological oxidation. Since then, millions of dollars have been placed into the

marketing and consumption of antioxidants (Harvard School or Public Health, 2014).

Before analyzing the supporting evidence of the biological benefits of antioxidants,

the terms free radical, oxidative stress, and antioxidants must be defined.

The general definition of a free radical is a molecule that has an unpaired

electron. Typically organic molecules contain an even number of electrons with each

electron displaying a magnetic spin that is opposite of its partner. Free radicals have

only one electron in their valence shell, and therefore this electron will have only

one directional spin. The single electron makes the free radical a very unstable

molecule.

In Biological free radicals, the main molecule involved is oxygen (O2). O2

contains two unpaired electrons and by definition should be a very reactive

compound; however, because both of the electrons of oxygen have the same

directional spin the compound remains relatively stable. If oxygen accepts another

electron (reduced) then the molecule contains only one unpaired electron, which is

highly unstable (fblt.cz, 2014). This produces a molecule called superoxide.

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Superoxide is a free radical, as well as one of many compounds called Reactive

Oxygen Species (ROS). Superoxide is naturally formed in the body inside the

mitochondria of cells. The electron transport chain and the coenzymes involved can

reduce oxygen and produce superoxide (Gropper & Smith, 2013). Superoxide has

the ability to help fight bacteria in the body, but it also has the ability to aid in the

production of other ROS compounds. If another electron is added to superoxide than

a compound called Hydrogen Peroxide (H2O2) is produced. Hydrogen Peroxide is not

a free radical, but it is an ROS. It has the ability to diffuse across cell membranes

and cause damage (Gropper & Smith, 2013). If Hydrogen Peroxide is able to react

with superoxide, a compound called a hydroxyl radical is produced. A hydroxyl

radical is the most powerful of free radicals and is also considered an ROS. Hydroxyl

radicals will quickly begin to attack molecules throughout the body by taking

electrons. Hydroxyl radicals are most commonly the cause of well-known biological

damage caused by free radicals such as: denaturing of proteins, DNA damage, and

lipid peroxidation. All of which can lead to more extensive diseases and illnesses.

Oxidative stress is the buildup of these ROS compounds. It can be caused by

pollution, radiation, UV exposure, high levels of oxygen, and physiological

preservation of the body. The ROS compounds then begin to cause damage to the

molecules, cells and systems of the body. Oxidative Stress is not something that can

easily be managed in todays world. Small amounts of oxidative stress produced

naturally by our body can be managed by the bodys natural antioxidants; however,

with a universe that continues to accumulate oxidative stress sources, other means

of protection are necessary.

Antioxidants are chemical nutrients that are able to donate electrons. In

theory, antioxidants are able to give electrons to the free radicles and ROS that
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dont contain an even pair. By doing this the free radical or ROS becomes

inactivated and is unable to continue to damage cells. Not all antioxidants are the

same, and they have individual properties and chemical makeup. Based off of the

theory that antioxidants are able to replenish electrons there have been much

experimental research done to validate the use of supplemental antioxidants.

A majority of the research that has been completed and supportive of the

beneficial use of antioxidants has been performed in vitro. This means that little

supportive evidence of antioxidants has been found using live human models. The

majority of testing that has occurred using human models indicated no inhibition of

ROS production or destruction. There is evidence that foods containing antioxidants

such as vegetables and fruits may be related to lower incidences of illness and

disease; however, there is a possibility that this is merely a correlation and not a

causation. This correlation could also be the result of high levels of other beneficial

nutrients such as phytochemicals (Ristow, 2014). In some studies using a large

number of test subjects the results indicated that not only do antioxidants cause

very little biological benefits, but they may even cause negative effects on the body.

There are several possible reasons as to why the results of these experiments do

not favor the beneficial use of antioxidants, but there is one reasoning that not only

explains these results, it also exposes the possibility of ROS producing beneficial

effects in the body. This process is called mitochondrial hormesis.

According to Ristow, ROS-the main target of antioxidants-either minimally

effect disease progression or may even help in some way to reduce the incidence of

chronic disease by promoting protective mechanisms within the cell under certain

circumstances (2014). One study as cited by Ristow, involve the caloric restriction

of Caenorhabditis elegans. When scientists decreased the caloric intake of

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Caenorhabditis the result was a buildup of ROS within the organisms and a decrease

in mortality rates. When antioxidants where given to the organisms in combination

with decreased calorie intake the results revealed that mortality rates increased.

This experiment and those with similar results could be because of a physiological

response called mitochondrial hormesis. Mitochondrial hormesis is a defense

mechanism that protects cells from damage termed to explain why small amounts

of ROS cause beneficial results but increased amounts can be destructive. This kind

of U-shaped response fosters the idea that ROS in small amounts will function as

signaling molecules. This response works in a very similar fashion as an immune

system/response in humans. The mitochondria are the primary location for the

synthesis of ROS. When the body undergoes different forms of oxidative stress the

mitochondria reacts by producing more ROS compounds. This then can trigger a

retrograde response that essentially communicates with the nucleus of cells to

undergo transcription of mRNA. This process produces more antioxidant enzymes;

more detox enzymes, and other compounds that prevent further oxidation of cells

(Ristow, 2014).

In some cases such as starvation or energy depletion, the mitochondria of

cells will produce more ROS to create a signal for the body to begin autophagy.

Autophagy is a term that means self-digestion (Gibson, 2014). When production of

ROS increases the damaged proteins, organelles, and other cellular structures can

undergo autophagy so that they can rid the body of the damage. This is a way for

the body to respond to its own internal alarm system, and provide the body with a

long-term protective shield (Hiroyuki & Finkel, 2014). With the knowledge of ROS

in the body working as a signaling system and producing defensive biological

mechanisms it suggests a possible cause as to why antioxidants have not been

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successful in past experiments. In theory, antioxidants inhibit the production of ROS

and the destruction caused by ROS. If antioxidants inhibit the protective aspects of

ROS than this offset of good and bad ROS levels could be the reason antioxidants

research is indicating either little or negative effects of antioxidants and suggests

that antioxidants may be preventing both the destructive and beneficial ROS from

working and in the end hindering the body from naturally resisting oxidative stress.

It is important to not forget that the effects of ROS employ a J-shaped

reaction and that high amounts of ROS in the body will damage cells and tissues.

Due to the negative effects of oxidants there is still continued experimental work

being done to vindicate the use and reputable benefits of antioxidants. The results

of these studies have indicated that there could potentially be some effect on

oxidative stress when exposed to specific antioxidants. According to one study a

mouse brain exposed to prions led to neurodegeneration and synthesis of ROS. The

brain was then exposed to antioxidant therapy using the antioxidant glutathione.

This antioxidant was used to inhibit the effects of NOX2 which is a part of an

enzyme system that contributes to the production of ROS. By inhibiting this enzyme

system there was neural protection (Hiroyuki & Finkel, 2014). Although the

antioxidant was found to be successful in this study, the beneficial effects began to

diminish as time increased. This study revealed that the benefits of antioxidants

may only be initially detected, and they must be used in a timely manner (in

relation to the cause of the ROS production). There was a second study focused on

examining the effects of glutathione and oxidative tissue damage of drosophillia.

The study was first performed in vitro and then again using live drosophilia. When

the study concluded, the in vitro experimental results indicated that glutathione

affects redox reactions within the cellular mechanism of the tissues, but when the
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study was done on live drosophila the same results were not found (Hiroyuki &

Finkel, 2014). This could mean that although glutathione plays a role in the redox of

cellular metabolism, this change found in the in vitro subject could be due to other

causes. With some evidence that does suggest that antioxidants do take part in the

protection against large amounts of ROS there must be recognition that when

applied correctly antioxidants could be influential in the future. It is important to

recognize that the studies that have supported antioxidants have been done on an

independent subject or in vitro, and that large group testing has not provided the

same results.

With conflicting results its hard to determine if antioxidants should be

encouraged, discouraged, or left to individual opinion. There are some factors that

should be considered when comparing results. The first is the size of the group

being tested. If a large group is studied the results appear drastic and definite due

to the large number of participants. However, a large group study is difficult to

control, and it is impossible to record the environment, health, and living quality of

every individual test subject. These minor influences could be the reason for a sway

in data. On the other hand, small study groups or individuals do not provide strong

statistical data, and individual results could dramatically affect the studys

conclusion (Unite For Sight).

There have been many studies on the benefits of antioxidants that have been

performed using large groups. A meta-analysis was completed to analyze the use of

supplemental vitamins and antioxidants and the effects on preventing

cardiovascular health using 324,653 subjects (Ristow M. , 2014). There was another

study completed using nearly 5,000 elderly people to determine the effect of

antioxidants such as vitamin C and E on the prevention of dementia (Ristow M. ,

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2014). The results of both of these studies indicated that antioxidants did not

provide beneficial protection against the identified diseases. With such large test

groups an experiment with complete accuracy is unattainable. The same can be

said for many of the individual and in vitro studies. The small studies that support

the use of antioxidants should be considered statistically insignificant, but often

times they provide distinguished results, and are used to produce profit rather than

definitive scientific data.

Another factor that could be affecting the experimental results of these

studies is the broad use of antioxidants to determine the effect of a very distinct

ailment. The study conducted in the use of antioxidants to prevent dementia used a

number of well-known antioxidants, but no results were found. The lack of results

may be caused by the functionality of the antioxidants tested. All antioxidants are

different in composition and function. For example, some antioxidants that

specifically attack free radicals and ROS in the body, while others attack enzymatic

complexes that catalyze the production of free radicals (Lobo, Patil, Phatak, &

Chandra, 2010). It is possible that because the functionality of antioxidants is not

fully understood their effects on health and disease cant be exclusively concluded

either. Currently, there is not a clear answer as to what method people should use in

preventing oxidative damage. The evidence that low levels of ROS can be beneficial

to the body is not strong enough to publicly endorse, and even if it was, the amount

of ROS in the body must be carefully maintained as large amounts are lethal to the

body. Antioxidants will continue to be pushed by manufacturers, but there is little

concrete evidence that proves they are beneficial to the body.

According to Finkel and Hiroyuki the best option may not be to prevent

oxidation, but instead to remove the damage that has been caused by the ROS.
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Based off of the evidence that ROS activates a response that initiates mitochondrial

autophagy it may be possible to expose cells to additional autophagy to reduce the

damage caused by oxidation (Hiroyuki & Finkel, 2014). The drug Rapamycin is often

given to people who have received tissue transplants. The drug has recently been

found to inhibit mTOR. The enzyme mTOR is a kinase that regulates the growth and

survival of cells. Researchers now believe that it may also play a role in the flux of

mitochondrial oxidation and the stimulation of autopagy (Cunningham, et al., 2007).

With this discovery researchers have continued to investigate the benefits of

Rapamycin in relation to oxidative stress, and have found when experimental mice

were given Rapamycin the life span of the mouse increased (Helmholtz Association

of German Research Centres, 2013). These new findings are still underdeveloped,

but the findings suggest that there may be future success in the pharmaceutical

approach to reduce the damage caused by oxidation.

For now, it is best for people to do what they can to reduce their exposure to

oxidative stress. People can certainly continue to promote the use of antioxidants,

but its important to recognize that the resolution to oxidative damage may be

dependent on a balance of ROS.

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