Christina Webb

Omega-3 and Inflammation

October 22,2013

Omega-3s Alone Are Not a Therapeutic Solution to Inflammation

Introduction
According to the CDC, the leading causes of death in the US include
Cardiovascular Disease (CVD), stroke, cancer and diabetes (Hoyert et. al,
2012). CVD, stroke, diabetes, and certain cancers like colitis are associated
with a high inflammatory state of the body. Inflammatory responses occur
when the body is exposed to stresses such as normal physiological stimuli or
injury. The bodys immune system is designed to produce inflammatory
responses for a short period of time to resolve the area of damage. In order
to prevent the body from damaging and digesting itself, inflammation is
tightly regulated. Inflammatory responses are often incurred through
signaling molecules, eicosanoids. Eicosanoids can be derived from essential
dietary PUFAs, omega-3 and omega-6. Omega-3s are considered antiinflammatory and omega-6s are generally considered inflammatory. The
American diet is rich in omega-6 and low in omega-3 with a ratio of about
16:1 respectively (Simopaulos, 2002). However, the optimal diet should
contain a ratio of four omega-6 to every one omega-3 (Simpaulos, 2002).
Considering the leading causes of death are due to inflammatory diseases
and inflammation related cancers, Omega-3 is currently being researched as
therapeutic solution to reduce inflammation.

Essential Fatty Acids

Omega-3: Omega-3s are a type of polyunsaturated fat that is essential
because the body cannot desaturate bonds after the delta 9 carbon. Marine

life, specifically cold water fish, are good sources of the Omega-3s,
Eiscosapentaenoic Acid (EPA) and Docosahexaenoic Acid(DHA). EPA (22:5)
can be converted into DHA via the Sprechers Shunt or can be oxidized by
enzymes cyclooxygenase (COX), lipoxygenase (LOX), and 5-LOX to produce
immune responses (Calder, 2010). COX is known to convert EPA into 3-series
prostaglandins (PG), which promote a low inflammatory response, and Eseries resolvins, which induce low anti-inflammatory responses and
inflammation resolving (Calder, 2010).

LOX is responsible for EPAs

conversion to E-Series resolvins. 5-LOX is responsible for EPAs conversion to

5-series Luekotrienes (LT) which act to promote a low pro-inflammatory state
(Calder, 2010). DHA (22:6) can be oxidized by COX and LOX. The COX and
LOX production from DHA results in D-series resolvins and protectins which
induce a potential anti-inflammatory and inflammation resolving state
(Calder, 2010). The Omega-3 Alpha-Linolenic Acid (ALA) is found in an
abundance of plants and plant oils. However, ALA (18:3) is the least
biologically active form of omega-3s because ALA has a 9% conversion rate
to EPA.
Omega-6: Omega-6 is a polyunsaturated fatty acid that is considered
essential because the human body cannot desaturate bonds after the delta 9
carbon. Linoleic Acid (LA) and Arachidonic Acid (AA) are common dietary
sources. Plant oils and plants such as avocados contain LA. LA is often
converted into AA. AA is consumed via animal sources, such as meat.
Omega-6 generates a series of reactions catalyzed by COX, 15-LOX, 12-LOX,

and 5-LOX. COX acts on AA, resulting in 2-series PGs such as PGd2, PGE2,
PGI2, PGF 2-alpha, and thromboxane A2 (Calder, 2010). 5-LOX produces 4series LTs LTE4 and LTB4 and produces 5-oxoETE. 12-LOX results in the
production of 12-hydroxyeicosatetranenoic acid (12-HETE) (Calder 2010).15LOX results in 15-HETE and Lipoxin A4 (Calder, 2010). All series of reactions
generate an environment conducive to high inflammatory reactions with the
exceptions of PGE2, thromboxane A2, and lipoxin A4 production (Calder,
2010).
COX Specificity
The type of stimulus triggering an immune response determines which set of
enzymes are utilized for eicosanoid production. For example, COX 1 is
triggered by normal physiological stimuli. COX-1 metabolizes AA to become
a certain prostanoids in the PGH2 family. PGH2 can be converted into
converted into thromboxane A4 which is responsible for decreasing platelet
aggregation (Greene et al, 2011). PGH2 can also be converted into PGE2
which is responsible from mucus secretion in the stomach (Greene et. al,
2011). Other prostanoids produced by COX 1 are also considered protective.
COX2 activity is stimulated when an infection or injury occur. COX2
metabolizes AA into inflammatory prostaglandins of the PGG2 family, which
results in a low pro-inflammatory state via the production of thromboxanes
and 3-series prostaglandins (Calder, 2010). These pro-inflammatory
prostaglandins and thromboxanes are responsible for innate immunity

responses such as pain, fever, and inflammation. The COX 2 enzyme is also
shown to inhibit apoptosis and promote cell proliferation (Zarghi et al, 2011).
Literature on PUFAs involvement in inflammation
Hurst et. al utilized cell cultures from individuals with osteoarthritis to study
the effects of omega-3 and omega-6 on COX-2 expression (Hurst et. al,
2009). Hurst and colleagues injected group one human cell cultures with 30
micrograms per mL of omega-3 and group 2 with 30 micrograms per mL of
omega-6. Later, the explants were washed and then injected with 10
nanograms per mL of interleukin-1 beta for human cultures 10 nanograms
per mL of interleukin-1 alpha for bovine cultures. After the injection, RNA
was extracted and analyzed using the PCR. A control group was maintained
for reference. Group 1 and Group 2 cultures injected with interleukin- 1 beta
had increased COX 2 levels. However the cells cultured with EPA had
significantly lower levels of COX 2 compared to the omega-6 group. Overall,
a decrease of COX-2 expression was seen when EPA was injected.
Gravaghi et. al studied the effects of endogenous omega-3 on colitis in
transgenic mice with colitis who express the C. elegans fat 1-gene vs mice
without the C. elegans fat 1-gene (Gravaghi et. al, 2011). C. elegans fat 1gene allows the mice to endogenously synthesize omega-3s. The mice were
categorized into three groups: control acute colitis, and chronic colitis each
fed a standard diet along with omega-6 diet via safflower oil for 7 weeks.
During the last week the mice were given a water solution containing 3%
DSS for 5 days to induce acute and chronic colitis. The mice were sacrificed

at 9 weeks and the colons were frozen and sectioned for examination.
Gravaghi et. al found the transgenic mice expressing the fat 1-gene not only
increased the omega-3 concentration in colonic cells compared to mice
without the gene, but also decreased the omega-6 concentration in colonic
cells. Furthermore, mice with the fat 1-gene showed decreased
inflammatory responses and changes to colon tissue when compared to their
non fat 1-gene counterparts. These differences are likely due to the
increased PGE2 expression and the decreased COX 2 levels in colon tissue.
Overall, endogenous synthesis of omega-3s reduced the inflammatory
response than the mice without the fat 1-gene.
Hyung-Wook and colleagues weaned Fischer-344 rats from their mothers and
were divided into two groups to study the omega-6 effects on COX activity
(Hyung-Wook et. al, 2011). Group 1, the control, was fed a diet adequate in
omega-3s and omega-6s (52.1 micromols per gram). Group 2, the
experimental group, was fed a diet adequate in omega-3s and inadequate
omega-6s at 4.2 micromols per gram of LA. At 15 weeks the rats were
sacrificed and their brains were extracted and preserved. Hyung-Wook et. al
analyzed mRNA and protein levels for COX-1 and COX-2 in the brains of both
groups. Overall, Group 2 saw significantly lower levels of mRNA COX-2 than
the control group, while COX-1 was not affected in either category.
Tull et. al used endothelial cell cultures injected with tumor necrosis factoralpha and human blood neutrophils in order to determine the effects of EPA

on mRNA expression of inflammatory factors (Tull et. al, 2009). The cells
were separated into two groups, the control group, which had no EPA infused
into the cell cultures, and the experimental group, which had 5 micromols of
EPA infused into the cell cultures. RNA was extracted and PCR was used to
analyze the differences in the cultures. The control group and the
experimental group had insignificant differences in the expression of
adhesion molecules, chemokines, and cytokines. However, Tull and
colleagues found tumor necrosis factor- alphas role in neutrophil migration is
inhibited with EPA but not when AA and EPA are added to the cultures.
Opinion
Omega-3 plays a role in COX-1 and COX 2 activity and thus effects
expression of 2-series prostaglandins and 3-series prostaglandins, both of
which have varying potential for triggering low anti-inflammatory and high
anti-inflammatory physiological states in the body. COX enzymes act on both
Omega-6 and Omega-3 PUFAS and seem to have a systemic immune effect.
Furthermore, Omega-3 is also known to play a role in the expression of
several other complex anti-inflammatory responses. However, the effects of
Omega-3 seem to be dependent on the dose and type of omega-3 the
eicosanoid is derived from (Rees et. al, 2006; Calder, 2010; Tull et. al, 2009).
Thus, omega-3s slightly decrease the overall state of inflammation (Refer to
Figure 1). Therefore, quality Omega-3s, such as DHA and EPA, should be
supplemented in the diet according the RDA. However, Omega- 3 intake

should not be seen as a therapeutic solution to inflammation due to its less

biologically active potency (Calder, 2010).

Discussion
Gravaghi et. als study does not measure exogenous Omega-3, which is the
only way humans can obtain Omega-3s, so the effect of Omega-3s may be
significantly altered by the physiological ability to produce Omega-3s
(Gravaghi et. al, 2011). Thus, endogenous Omega-3s may play a different
role in the genetic expression of inflammatory factors. Furthermore, despite
Omega-3 affecting the activity of COX enzymes, it appears omega-6 intake
may play a larger role in the expression of high inflammatory agents (Calder,
2010; Tull et. al, 2009). Because Omega-6 is more biologically potent than
EPA and because omega-6 has protective and inflammatory effects, it is
important to balance out the diet with the correct omega-6:omega-3 ratio.
Thus, diet therapy of reduced omega-6 and increased omega-3 may play a
larger physiological role in inflammation than Omega-3 alone. More studies
should be conducted in animal and human models to determine the effects
of omega-6:omega-3 ratio in a normal state and an inflammatory state to
determine the therapeutic value.

Figure 1: Dietary Omega-3s Role in

Inflammation

Citations
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