-Tocopherol, but not -tocopherol, decreases
proinflammatory eicosanoids and inflammation
damage in rats
QING JIANG AND BRUCE N. AMES1
Division of Biochemistry and Molecular Biology, University of California, Berkeley; and Children's
Hospital Oakland Research Institute, Oakland, California, USA
ABSTRACT -Tocopherol (T), the major form of
vitamin E in U.S. diets, and its physiological metabolite
2, 7, 8-trimethyl-2-(-carboxyethyl)-6-hydroxychroman
(-CEHC), in contrast to -tocopherol (T), the pri-
mary vitamin E in supplements, inhibit cyclooxygenase-
catalyzed synthesis of prostaglandin E2 (PGE2) in acti-
vated macrophages and epithelial cells. Here we report
that in carrageenan-induced inflammation in male
Wistar rats, administration of T (33 or 100 mg/kg)
and -CEHC (2 mg/pouch), but not T (33 mg/kg),
significantly reduced PGE2 synthesis at the site of
inflammation. T, but not T, significantly inhibited
the formation of leukotriene B4, a potent chemotactic
agent synthesized by the 5-lipoxygenase of neutrophils.
Although T had no effect on neutrophil infiltration, it
significantly attenuated the partial loss of food con-
sumption caused by inflammation-associated discom-
fort. Administration of T led consistently to a signifi-
cant reduction of inflammation-mediated increase in
8-isoprostane, a biomarker of lipid peroxidation. T at
100 mg/kg reduced TNF- (65%;P0.069), total ni-
trate/nitrite (40%;P0.1), and lactate dehydrogenase
activity (30%;P0.067). Collectively, T inhibits proin-
flammatory PGE2 and LTB4 , decreases TNF-, and
attenuates inflammation-mediated damage. These find-
ings provide strong evidence that T shows anti-inflam-
matory activities in vivo that may be important for
human disease prevention and therapy.Jiang, Q.,
Ames, B. N. -Tocopherol, but not -tocopherol, de-
creases proinflammatory eicosanoids and inflammation
damage in rats. FASEB J. 17, 816 822 (2003)
Key Words: vitamin E -tocopherol metabolite prostaglan-
din E2 leukotriene B4 TNF-alpha
Inflammatory diseases such as rheumatoid arthritis,
asthma, and hepatitis are among the leading causes of
death and disability in the world. Chronic inflamma-
tion contributes to the development of degenerative
diseases, including cancer (1), cardiovascular diseases
(2), and neurodegenerative disorders (3). During in-
flammation, various eicosanoids derived from arachi-
donic acid (AA) play a key role in mediating inflamma-
tory response (4). For instance, prostaglandin E2
(PGE2), which is synthesized from cyclooxygenase
816
(COX)-catalyzed oxidation of AA, is believed to cause
pain and fever (4, 5), as well as activate cytokine
formation (6). PGE2 can be produced by either the
constitutive form (COX-1) or the inducible form
(COX-2) of cyclooxygenase (7). In most inflammatory
conditions, COX-2 is up-regulated and is the primary
enzyme responsible for the formation of proinflamma-
tory PGE2 (7). Leukotriene B4 (LTB4), another oxi-
dized product derived from AA through the 5-lipoxy-
genase-catalyzed pathway, is one of the most potent
chemotactic agents (8). Because of the central roles of
PGE2 and LTB4 , COX-2 and 5-lipoxygenase have been
recognized as key targets for drug therapy in inflamma-
tion-associated diseases. In particular, COX-2 inhibi-
tors, which are classified as nonsteroidal anti-inflamma-
tory drugs (NSAIDs), have proved effective in
attenuating inflammatory response and are beneficial
for certain inflammation-associated diseases (9).
We recently found that -tocopherol (T), the major
form of vitamin E in the U.S. diet, and its physiological
metabolite, 2, 7, 8-trimethyl-2-(-carboxyethyl)-6-hydroxy-
chroman (-CEHC), inhibit COX-2-catalyzed formation
of PGE2 , as assayed in lipopolysaccharide-stimulated
macrophage and interlukin-1-activated epithelial cells
(10). This indicates that T and its metabolite may have
anti-inflammatory properties similar to those of
NSAIDs. In contrast, -tocopherol (T), the predomi-
nant form of vitamin E in the tissues and most supple-
ments, is much less effective in this regard (10). The
present study is aimed to test whether T and its
metabolite inhibit PGE2 and other proinflammatory
eicosanoids in an air pouch model where inflammation
is induced by a single injection of carrageenan (11).
For comparison, the effect of T was tested. In addition
to eicosanoids, the effects of T on the generation of
tumor necrosis factor- (TNF-), an inflammatory cy-
tokine, reactive nitrogen oxide, and oxidative damage
were also investigated.
1
Correspondence: CHORI, 5700 Martin Luther King Jr.
Way, Oakland, CA 94609-1673, USA. E-mail: bnames@
uclink4.berkeley.edu
0892-6638/03/0017-0816 FASEB
MATERIALS AND METHODS
Materials
T (99%) and T (95%-97%) were purchased from Acros
Organics (Fairlawn, NJ, USA) or Fluka (Milwaukee, WI,
USA). -CEHC (98%) was from Cayman Chemicals (Ann
Arbor, MI, USA). Carrageenan, -tocopherol acetate, and all
other chemicals were from Sigma (St. Louis, MO, USA).
Carrageenan-induced inflammation in the air pouch model
The animal use protocol was approved by the animal care
committee at the Children's Hospital Oakland Research
Institute and strictly followed. Male Wistar rats (250 330 g)
(Charles River, CA, USA) were caged singly and routinely fed
ad libitum with Purina Chow with free access to tap water. An
air pouch was created by a subcutaneous injection of 12 mL
sterile air into the intrascapular area of the rat's back. Thirty
hours later, 2 mL of 0.5% carrageenan or phosphate-buffered
saline (PBS) (controls) was injected into the air pouch. To
test the effect of -CEHC, which is water soluble, 2 mg per
pouch was dissolved in PBS and injected directly into the
pouch right before carrageenan injection. Six hours after the
induction of inflammation, animals were killed and the
pouch fluid was collected to evaluate the inflammatory re-
sponse (see below). In the experiments with tocopherols,
animals were fed with T (33 or 100 mg/kg) or T (33
mg/kg), continuously for 3 days by gavage using 0.5 mL
tocopherol-stripped corn oil (Dyets Inc., Bethlehem, PA,
USA) as the vehicle, before injection of carrageenan. Control
animals received the same volume of tocopherol-stripped
corn oil. Twenty hours after the injection of carrageenan
(0.5%, 2 mL), rats were killed and pouch fluid was collected
by lavage with Hanks-buffered saline solution containing
0.004% heparin but no Ca2/Mg2. After a brief centrifuga-
tion, the supernatant was collected and frozen immediately
on dry ice for the measurement of PGE2 , LTB4 , and TNF-,
etc. Total cells were counted by a Coulter counter or hemo-
cytometer.
Measurement of T and T
Plasma and exudate T and T were extracted using a
mixture of methanol/hexane (2:5, v/v) in the presence of 0.8
mM butylated hydroxytoluene (BHT) (12). After brief cen-
trifugation at 4C, the top hexane layer was dried under N2
and the residue was resuspended in ethanol. Tocopherols
were separated on a 150 4.6 mm, 5 m Supelcosil
LC-18-DB column (Supelco, Bellefonte, PA, USA) and eluted
with 95:5 (v/v) methanol/0.1M lithium acetate (final 25 mM,
pH 4.75) at a flow rate of 1.3 mL/min. Tocopherols were
monitored by coulometric detection (Model Coulochem II,
ESA Inc., Chelmsford, MA, USA) at 300 (upstream) and 500
mV (downstream electrode) using a Model 5011 analytical
cell.
Quantification of -CEHC
The sample preparation procedure was modified from a
published method (13). Briefly, 200 L of plasma was diluted
into the same volume of cold PBS with 5% ethanol and the
lipid components were removed by an extraction with 0.5 mL
hexane containing 0.8 mM of BHT. After aspiration of the
hexane layer, the remaining aqueous phase was acidified to
pH 3 4 with acetic acid. -CEHC was then extracted twice
with 1 mL ethyl acetate containing 0.8 mM BHT. The
IN VIVO ANTI-INFLAMMATORY EFFECTS OF -TOCOPHEROL
combined ethyl acetate layer was evaporated under N2 and
the residue was resuspended in the HPLC elute solvent (see
below). -CEHC was injected onto a 150 4.6 mm, 5 m
Supelcosil LC-18-DB column (Supelco) and eluted with 10
mM lithium acetate (pH 4.3) containing 35% acetonitrile at a
flow rate of 1.0 mL/min. -CEHC was monitored by coulo-
metric detection (Model Coulochem II, ESA Inc.) at 300 mV
using a Model 5011 analytical cell.
Measurement of PGE2 , LTB4 , 8-isoprostane, LDH,
and TNF- in the exudate
For measurement of PGE2 , LTB4 , and 8-isoprostane, the
exudate fluid was mixed vigorously with 2 mL methanol to
precipitate proteins and with 5 mL hexane to remove lipids.
After a brief centrifugation and aspiration of the hexane
layer, the methanol layer was removed and evaporated under
N2. PGE2 , LTB4 , and 8-isoprostane were measured using the
corresponding ELISA kits from Cayman Chemicals. TNF-
and lactate dehydrogenase (LDH) in the exudate were mea-
sured directly using an ELISA kit from R&D (Minneapolis,
MN, USA) and an analytical kit from Roche (Indianapolis, IN,
USA), respectively.
Quantitation of total NO2- and NO3-
The total amount of nitrate and nitrite in the exudate was
measured using the Model 280 nitric oxide analyzer
(NOATM) (Sievers Instruments, Inc., Boulder, CO, USA).
Nitrite and nitrate in the exudate were reduced by vanadium
(III) to nitric oxide, which was then measured by a red-
sensitive photomultiplier tube after being converted by ozone
to chemiluminescent reactive nitrogen dioxide. Nitrite and
nitrate were quantified based on a standard curve from
nitrate established under identical conditions.
Statistics
A nonpaired Students t test was performed in all the data
analyses. Data are expressed as mean se.
RESULTS
Administration of T and its major metabolite,
-CEHC, but not T, significantly inhibited
proinflammatory eicosanoids at the site of
inflammation
Carrageenan-induced inflammation in the air pouch
model is commonly used to evaluate the pharmaceuti-
cal potency of anti-inflammatory drugs (14). In this
model, an injection of air into the intrascapular area
resulted in the formation of a connective tissue cavity
lined mainly with macrophages and fibroblasts (11, 15).
These cells play a key role in the inflammatory response
(11, 14, 15). A single injection of carrageenan caused a
potent localized inflammation as indicated by a marked
increase in white cell infiltration, eicosanoid formation,
and tissue damage (11). To study the effect of -CEHC
on eicosanoid synthesis and neutrophil infiltration, it
(1 mg/mL, 2 mL in PBS) was injected directly into the
air pouch right before the injection of carrageenan.
Since the retention of -CEHC is relatively short (16),
817
Figure 1. Effects of -CEHC on PGE2 , LTB4 ,
and neutrophil infiltration. In the air pouch
model, 2 mL of 1 mg/mL -CEHC, or 2 mL
PBS (vehicle) was injected into the pouch
right before injection of 2 mL, 0.5% carra-
geenan to initiate inflammation. In the con-
trols for inflammation, PBS was injected
instead of carrageenan (Ctrl). 6 h after
carrageenan injection, the effects on PGE2
(A), LTB4 (B), and neutrophil infiltration
(C) were evaluated. *(P0.05) indicates a
statistically significant difference between
-CEHC-administrated rats (n8) and PBS-
injected animals (n7).

its effect was evaluated 6 h after the induction of
inflammation. Administration of -CEHC significantly
reduced PGE2 (29%, P0.05) in the pouch (Fig. 1A),
which is consistent with our previous observations in
vitro (10). At this dose, -CEHC also inhibited LTB4
and neutrophil infiltration (Fig. 1B, C), though not
significantly.
To test the effects of tocopherols, T or T dissolved
in tocopherol-stripped corn oil was continuously ad-
ministrated by gavage for 3 days before the induction of
inflammation. Effects were evaluated at 20 h after
carrageenan injection, when cell infiltration had
reached a maximum in the pouch (data not shown). At
a dose of 33 mg/kg, T, but not T, significantly
reduced the proinflammatory PGE2 (46%, P0.05; Fig.
2A) and LTB4 (70%, P0.05; Fig. 2B), a potent chemo-
tactic eicosanoid produced by the 5-lipoxygenase in
neutrophils. At a higher dose (100 mg/kg), T showed
an inhibitory potency against PGE2 (51%, P0.05) and
LTB4 similar to the lower dose. Despite the inhibitory

Figure 2. Effects of T and T on PGE2 , LTB4 ,

and neutrophil infiltration. T at 33 mg/kg
(T33, n7) or 100 mg/kg (T100, n8) and
T at 33 mg/kg (T, n7) dissolved in corn oil
(0.5 mL) were administered by gavage contin-
uously for 3 days before carrageenan injection
(2 mL 0.5%). Rats administered with 0.5 mL of
corn oil for 3 days and injected with carra-
geenan served as vehicle controls (Corn, n8).
Control rats (Ctrl, n5) were those gavaged
with 0.5 mL corn oil and injected with 2 mL
PBS. 20 h after carrageenan injection, the
effects on PGE2 (A), LTB4 (B), and neutrophil
infiltration (C) were evaluated. *(P0.05) in-
dicates a statistically significant difference be-
tween tocopherol-treated rats and corn oil-
administrated animals.

818 Vol. 17 May 2003 The FASEB Journal JIANG AND AMES
effects on PGE2 and LTB4 , T did not affect neutrophil
infiltration (Fig. 2C).

T reduced TNF- and total nitrite and nitrate at the

higher dose of 100 mg/kg

Besides eicosanoids, we have investigated the effects of

T on TNF-, an important inflammation mediator,
and total nitrite and nitrate, an index of the generation
of reactive nitrogen oxides (Fig. 3). At the lower dose of
33 mg/kg, T nonsignificantly inhibited TNF-
(51%, P0.29) but had no effect on total nitrate and
nitrite. At the higher dose of 100 mg/kg, T reduced
TNF- (65%, P0.069) (Fig. 3A) and total nitrite and
nitrate (40%, P0.1) (Fig. 3B).

Administration of T attenuated the partial loss of

food consumption induced by inflammation and
inhibited inflammation-mediated lipid peroxidation
and cytotoxicity

The effect of T and T on inflammation-induced lipid

peroxidation and inflammation site tissue damage was
assayed by 8-isoprostane levels (17) and by the release
of LDH, respectively (11). Carrageenan-induced in-
flammation resulted in a marked increase in 8-isopros-
tane and LDH in the pouch. In contrast to T (33
mg/kg), T at the dose of 33 or 100 mg/kg significantly
reduced 8-isoprostane (for both doses, 57%, P0.05)
(Fig. 4A) in the pouch. At the higher dose of 100
mg/kg, T lowered LDH (30%, P0.067) (Fig. 4B), a

Figure 4. Effects of T and T on the formation of 8-isopros-

tane (A), LDH (B), and food consumption (C). The condi-
tions of each treatment are the same as indicated in Fig. 2.
The food (%) consumption of individual animal was the ratio
of food intake measured before and after the carrageenan
injection. *(P0.05) indicates a statistically significant differ-
ence between the tocopherol-treated rats and the corn oil-
administrated animals.

marker of cytotoxicity and tissue damage. Carrageenan-

induced inflammation resulted in a marked reduction
in food consumption (40%, P0.01), which is likely
caused by the discomfort associated with inflammation.
Administration of T at 100 mg/kg significantly atten-
uated (30%, P0.03) the loss of food consumption; a
smaller, nonsignificant effect was observed at the lower
dose of 33 mg/kg (20%, P0.2) (Fig. 4C).

Plasma and exudate concentrations of T, T,

Figure 3. Effects of T and T on the accumulation of TNF-
(A) and total nitrate/nitrite (B). The conditions of each
treatment are the same as indicated in Fig. 2. P values indicate
the statistical comparison between the tocopherol-treated
group and the corn oil-fed group (Corn).
and -CEHC
To evaluate the relative bioavailability of the adminis-
trated compounds, we measured plasma and exudate
concentrations of T and T as well as plasma -CEHC

IN VIVO ANTI-INFLAMMATORY EFFECTS OF -TOCOPHEROL 819

Figure 5. Plasma and exudate concentra-
tions of T and T and plasma concen-
trations of -CEHC. The conditions of
each treatment are the same as indicated
in Fig. 2. *(P0.05) or **(P0.01) and
a
(P0.01) indicate a significant differ-
ence between the tocopherol treated
group and corn oil-fed group.
(Fig. 5). Administration of T or T led to significant
increases in both tocopherols in the plasma and exu-
date, whereas their relative increase in the exudate was
more than that in the plasma. Thus, T-administrated
(33 or 100 mg/kg) rats had nearly 10- or 20-fold
elevation of T in exudate fluid, in contrast to 3- or
5-fold increase in the plasma compared with corn
oil-fed controls (Fig. 5). A similar trend was observed
with T. In T-administrated rats (33 or 100 mg/kg),
the ratio of T to T in the exudate (0.3 or 0.7) is
higher than that in the plasma (0.15), consistent with
the idea that tissues may have a higher T partition
than does plasma (18). T administration did not
significantly affect T, but T caused significant de-
creases of T in both the plasma (55%, P0.05) and
exudate (40%, P0.05). -CEHC, the major metabolite
of T, increased in response to T supplementation.
Nanomolar concentrations of -CEHC were found in
the plasma, a level 10% that of plasma T. T
administration caused a 2.5- to 4-fold elevation of
-CEHC in the plasma (Fig. 5C).
DISCUSSION
Carrageenan-induced inflammation in the rat air
pouch model is believed to mimic the pathological
process occurring in joint diseases such as arthritis. This
is because the connective tissues formed along the air
pouch are similar to those found in chronic joint
diseases (11, 15). Carrageenan-induced inflammation
and chronic joint diseases share other features, includ-
ing markedly elevated PGE2 , neutrophil infiltration,
cytokine formation, and tissue damage (11). Studies
have established that in this model, COX-2, which is
quickly induced in the lining macrophages and fibro-
820 Vol. 17 May 2003 The FASEB Journal
blasts, is the primary enzyme responsible for the eleva-
tion of PGE2 (14). Thus, various COX-2 inhibitors have
been shown to inhibit the formation of PGE2 in the
pouch (14). We recently found that the major form of
vitamin E in the diet, T, and its metabolite, but not T,
the major form in supplements, inhibited COX-2 activ-
ity in lipopolysaccharide-activated macrophages and
interlukin-1-treated epithelial cells (10). In line with
this in vitro observation, the present study shows that in
the carrageenan air pouch model, T (33 or 100
mg/kg), in contrast to T (33 mg/kg), significantly
lowered PGE2 elevation at the site of inflammation.
Local delivery of -CEHC into the air pouch also led to
a significant inhibition of PGE2. These results therefore
demonstrate that T and -CEHC show in vivo anti-
inflammatory properties that appear to be similar to
those of NSAIDs. T but not T significantly inhibits
LTB4 , a potent chemotactic agent that is synthesized by
5-lipoxygenase of neutrophils (8).
In addition to the inhibitory effects on the proinflam-
matory eicosanoids, in this model, T administration
reduced inflammation-mediated damage, as shown by
reduced lipid peroxidation and LDH activity. T atten-
uated the marked loss of food consumption that is
likely caused by inflammation-associated discomfort.
Because PGE2 is known to play a key role in causing
pain and fever, a reduction of this eicosanoid may
explain, in part, Ts effect on the food consumption.
Besides the inhibition of COX-catalyzed reaction, other
unique properties of T may contribute to the observed
beneficial effects (18). Because of the unsubstituted
5-position compared with T, T is capable of trapping
reactive nitrogen oxide, such as nitrogen dioxide (19,
20) and peroxynitrite (21), to form a stable nitrated
adduct. T is better than T in protecting peroxyni-
trite-induced lipid peroxidation (21) and enzyme inac-
JIANG AND AMES
tivation (22). We recently found that in the zymosan-
induced inflammation model, T supplementation
consistently inhibited protein nitration and ascorbate
oxidation (12).
At 100 mg/kg, but not 33 mg/kg, T lowered accu-
mulation of total nitrate and nitrite (Fig. 3B). Although
some studies show that reactive nitric oxide stimulates
PGE2 formation (23, 24), in the current model these
two events appear to be independent because T
decreases PGE2 at both doses. In LPS-stimulated mac-
rophages, we earlier found that T moderately inhibits
nitrite accumulation via a moderate inhibition of the
induction of inducible nitric oxide synthase (10). It is
possible that the currently observed reduction of total
nitrate and nitrite is caused by Ts suppression of this
enzyme. Alternatively, Ts ability to trap reactive nitro-
gen oxides may also result in lowered levels of total
nitrate and nitrite. However, we did not observe an
apparent increase in 5-nitro--tocopherol in the exu-
date fluid (data not shown). Nevertheless, this possibil-
ity cannot be ruled out due to not measuring 5-nitro-
-CEHC, a putative breakdown product of 5-nitro--
tocopherol (18).
In this model T appears to decrease TNF- (Fig.
3A), a key proinflammatory cytokine known to activate
macrophages and provoke the inflammatory response
(25). Studies have shown that inhibition of TNF-
provides beneficial effects on inflammatory diseases
(26). Treatment with an antibody against TNF- has
been proved to be an effective therapy for inflamma-
tory disease (27). We do not know the mechanism
behind Ts inhibition of TNF-.
Although T significantly inhibits the proinflammatory
eicosanoids, it has no effect on neutrophil infiltration. It
has been shown that in the carrageenan air pouch model,
there is no casual correlation between the inhibition of
PGE2 and neutrophil infiltration (11). For example, aspi-
rin at doses of 100 150 mg/kg caused 5070% reduction
of PGE2, yet did not affect neutrophil infiltration (28).
Although at higher doses (200 300 mg/kg) aspirin
inhibits cell infiltration, the mechanisms may include
inhibition of NF-B signal transduction (29) or the acti-
vation of adenosine formation (30).
The relatively high bioavailability of T contributes to
its in vivo inhibition of eicosanoids in the pouch. T
administration resulted in a more pronounced elevation
of this tocopherol in the exudate than that in the plasma
(Fig. 5). When T showed significant inhibitory effects on
PGE2 at 20 h after the injection of carrageenan, its
concentration in the collected exudate was estimated to
be 93.3 (33 mg/kg) to 193 nmol/g protein (100 mg/kg)
(Fig. 5B), corresponding to 10 to 20 M, which is
comparable to the apparent IC50 (510 M) estimated
in cell culture experiments (10). An increase in T
administration from 33 to 100 mg/kg did not significantly
improve the inhibitory potency, probably due to the
saturation effect. Although when applied directly into the
pouch, -CEHC significantly inhibited PGE2 , the effect of
T cannot be attributed to this metabolite. This is because
the concentration of -CEHC is in the nanomolar range
IN VIVO ANTI-INFLAMMATORY EFFECTS OF -TOCOPHEROL
(Fig. 5C), which is much lower than the estimated IC50
(30 40 M) for -CEHC to inhibit COX-2 activity (10).
Though as much as 50% of T may be converted to
-CEHC (31), this metabolite is not likely to be accumu-
lated in the plasma or tissues (except the kidney) because
of its short retention time (16).
Although administration of T (33 mg/kg) led to an
approximately twofold increased level in the plasma and
exudate, T showed no significant effects in the present
study with respect to the generation of eicosanoids and
8-isoprostane, in line with our in vitro observations (10). It
may be relevant that T has a high baseline level in tissues
as a result of the high content of T in the diet. However,
administration of the same dose of T did show signifi-
cant effects. This observation indicates that T possesses
unique properties that are not shared T (18). Because
T is preferentially retained by the body and is the
strongest antioxidant in the vitamin E family, the possibil-
ity that supplementation of T and T (especially when
T is relatively low) may result in better outcomes war-
rants more investigation.
The in vivo anti-inflammatory activity of T, as demon-
strated by the current study, may be important for human
disease prevention and therapy. 1) It may be that T can
be used as a supplement for treatment of inflammatory
diseases to decrease proinflammatory eicosanoids and
damage from inflammation. Not only does T reduce
PGE2 , it inhibits lipoxygenase-catalyzed synthesis of LTB4
and decreases TNF-, an activity most NSAIDs do not
have (28). These findings suggest a potentially superior
pharmaceutical use for T compared with traditional
NSAIDs. 2) T may be useful in cancer prevention. It is
known that COX-2 and PGE2 are elevated in inflamma-
tion-associated diseases, including cancer (32). Frequent
intake of NSAIDs such as aspirin is known to reduce the
risk of certain cancers (33, 34). Recently, Helzlsouer et al.
(35) reported that in a nested case control study, men in
the highest quintile of plasma T levels had a fivefold
reduction in the risk of prostate cancer compared with
those in the lowest quintile. We recently found that T but
not T showed antiproliferative effects on prostate and
lung cancer cell lines, but had no effect on normal
epithelial cells (Q. Jiang, unpublished observation). 3)
The anti-inflammatory effects of T may be beneficial in
cardiovascular disease, another inflammation-associated
disorder (36). Several studies (37, 38) reported that
plasma concentrations of T, but not T, are inversely
associated with the incidence of coronary heart diseases.
Frequent intake of nuts, a rich source of T, is associated
with reduced risk for cardiovascular disease (39).
In conclusion, T inhibits the proinflammatory eico-
sanoids, suppresses proinflammatory cytokine, and at-
tenuates inflammation-mediated damage in a rat in-
flammation model. The current observations together
with the cited studies strongly suggest that T, the most
abundant form of vitamin E in the U.S. diet, is impor-
tant to human health and deserves more attention.
We thank Emily Ho, Takashi Akiyama, and Helen Song for
help with some animal experiments and Sandra Larkin,
821
Lorenzo Machado, and Frans Kuypers for technical assistance in 20. Cooney, R. V., Harwood, P. J., Franke, A. A., Narala, K.,
the measurement of nitrite/nitrate. This work was supported in Sundstrom, A. K., Berggren, P. O., and Mordan, L. J. (1995)
part by fellowships (AHA98-24) from the American Heart Asso- Products of gamma-tocopherol reaction with NO2 and their
ciation-Western Affiliates and the Cancer Research Laboratory formation in rat insulinoma (RINm5F) cells. Free Radic. Biol.
Med. 19, 259 269
of the University of California at Berkeley (to Q.J.), the Wheeler
21. Christen, S., Woodall, A. A., Shigenaga, M. K., Southwell-Keely,
Fund of the Dean of Biological Science at the University of P. T., Duncan, M. W., and Ames, B. N. (1997) gamma-to-
California Berkeley, and the National Institute of Environmental copherol traps mutagenic electrophiles such as NO(X) and
Sciences Center Grant ES01896 (to B.N.A.). complements alpha-tocopherol: physiological implications.
Proc. Natl. Acad. Sci. USA 94, 32173222
22. Williamson, K. S., Prasad Gabbita, S., Mou, S., West, M., Pye,
Q. N., Markesbery, W. R., Cooney, R. V., Grammas, P., Reimann-
Philipp, U., Floyd, R. A., et al. (2002) The nitration product
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822 Vol. 17 May 2003 The FASEB Journal JIANG AND AMES