Fatty acid oxidation affects food intake by altering
hepatic energy status
MARK I. FRIEDMAN, RUTH B. HARRIS, HONG JI,
ISRAEL RAMIREZ, AND MICHAEL G. TORDOFF
Monell Chemical Senses Center, Philadelphia, Pennsylvania 19104
Friedman, Mark I., Ruth B. Harris, Hong Ji, Israel
Ramirez, and Michael G. Tordoff. Fatty acid oxidation
affects food intake by altering hepatic energy status. Am. J.
Physiol. 276 (Regulatory Integrative Comp. Physiol. 45):
R1046–R1053, 1999.—Inhibition of fatty acid oxidation stimu-
lates feeding behavior in rats. To determine whether a
decrease in hepatic fatty acid oxidation triggers this behav-
ioral response, we compared the effects of different doses of
methyl palmoxirate (MP), an inhibitor of fatty acid oxidation,
on food intake with those on in vivo and in vitro liver and
muscle metabolism. Administration of 1 mg/kg MP selectively
decreased hepatic fatty acid oxidation but did not stimulate
food intake. In contrast, feeding behavior increased in rats
given 5 or 10 mg/kg MP, which inhibited hepatic fatty acid
oxidation to the same extent as did the low dose but in
addition suppressed fatty acid oxidation in muscle and pro-
duced a marked depletion of liver glycogen. Dose-related
increases in food intake tracked dose-related reductions in
liver ATP content, ATP-to-ADP ratio, and phosphorylation
potential. The findings suggest that a decrease in hepatic
fatty acid oxidation can stimulate feeding behavior by reduc-
ing hepatic energy production.
feeding behavior; metabolism; liver; adenosine 58-triphos-
phate; methyl palmoxirate
A VARIETY OF EVIDENCE indicates that the oxidation of
fatty acids influences feeding behavior (see Ref. 4), with
perhaps the most direct support from experiments
showing that rats increase food intake after treatment
with agents that inhibit the oxidation of fatty acids (5,
24). Administration of either mercaptoacetate (MA),
which suppresses fatty acid oxidation by inhibiting the
acyl-CoA dehydrogenases that catalyze mitochondrial
␤-oxidation (see Ref. 24), or methyl palmoxirate (MP),
which reduces fatty acid oxidation by inhibiting the
intramitochondrial transport of fatty acids (1, 34, 35),
stimulates feeding behavior in rats (e.g., Refs. 5, 7, 18).
The effect of these inhibitors on food intake is most
evident when the maintenance diet is high in fat or
glucose utilization is impaired (7, 8, 27), that is, when
fats are a major source of metabolic energy. This
further suggests that a decrease in fatty acid oxidation
underlies the eating response to administration of MA
or MP.
The increase in feeding behavior seen after inhibition
of fatty acid oxidation appears to be triggered by a
signal generated in the liver, because total vagotomy,
visceral deafferentation induced by systemic capsaicin,
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R1046 0363-6119/99 $5.00 Copyright r 1999 the American Physiological Society
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or hepatic branch vagotomy prevents or attenuates the
response to MA (13, 20, 21). However, because MA was
given systemically in these denervation experiments, it
is unclear whether MA stimulates eating by inhibiting
the oxidation of fatty acids in liver or whether the
metabolic consequences of MA acting in other tissues
are detected by a hepatic sensor. In the experiments
described below, we attempted to resolve this question
by using MP to selectively inhibit fatty acid oxidation in
liver.
MP suppresses fatty acid oxidation by inhibiting
carnitine palmitoyltransferase I (CPT I), which trans-
ports long-chain fatty acids into mitochondria (1, 32,
34). This mechanism of action apparently underlies the
behavioral effect of MP, because provision of medium-
chain fatty acids, which do not depend on CPT I for
intramitochondrial transport and oxidation, prevents
the eating response (5). Presumably, because the liver
and muscle isoforms of CPT I have different affinities
for MP (12), low doses of MP markedly inhibit CPT I in
liver but have relatively little effect on CPT I in muscle
(32), the other major tissue that oxidizes fatty acids. We
took advantage of this differential sensitivity of liver
and muscle CPT I and compared the dose-response
relationships between the effects of MP on food intake
with those on metabolism to determine whether a
decrease in fatty acid oxidation restricted to the liver
stimulates eating in rats.
METHODS AND PROCEDURES
Animals and diets. Adult male CD Sprague-Dawley rats
(Charles River; Wilmington, MA) weighing 350–450 g at the
time of testing, were used for all experiments. Rats were
housed individually in temperature-controlled (22°C) rooms
that were maintained on a 12:12-h light-dark cycle (lights on
at 0700). Rats were fed a high-fat, low-carbohydrate diet with
63 and 13% of energy as fat (corn oil) and carbohydrate,
respectively (19), and given tap water ad libitum except when
noted otherwise. Rats were fed the experimental diet for
10–14 days before testing. Food intakes were measured to the
nearest 0.1 g, corrected for spillage.
Inhibitor treatment. MP (methyl 2-tetradecylglycidate) was
given to rats by gavage tube at 1–2 h after the start of the
daylight period. MP was suspended in a methyl cellulose
(0.05%) vehicle, and both MP and vehicle were given in a
volume of 3 ml/kg. MP suspensions were sonicated and mixed
by vortexing before gavage. Groups of rats receiving different
doses of MP (or vehicle) were matched on the basis of body
weight.
Blood and tissue assays. Blood was collected in heparinized
tubes either from the tip of the tail (for repeated sampling) or
from the trunk after decapitation. Plasma obtained after
centrifugation was analyzed for glucose using a Beckman
glucose analyzer (glucose oxidase) and for total ketone bodies
(␤-hydroxybutyrate plus acetoacetate), free (unesterified) fatty
 FATTY ACID OXIDATION AND FOOD INTAKE R1047

acids (FFA), triglycerides, and glycerol using enzymatic as-

says with fluorometric detection (see Ref. 16). Liver glycogen
was measured using an enzymatic procedure (10).
For measurement of liver ATP, ADP, and Pi, rats were
anesthetized by subcutaneous injection of ketamine (100
mg/ml) plus acepromazine maleate (1 mg/ml). A sample of the
median lobe of the liver was removed through a midline
incision, immediately freeze-clamped in aluminum blocks
cooled in liquid nitrogen, and then submerged in liquid
nitrogen. Samples were stored at ⫺70°C and then assayed for
adenine nucleotides by HPLC (11, 23) and for Pi using a
commercial kit (no. 360–3; Sigma, St. Louis, MO). Phosphory-
lation potential was calculated as [ATP/(ADP ⫻ Pi )].
In vitro fatty acid metabolism. Rats were decapitated, and
samples of liver tissue and diaphragm (muscle) were collected
through a midline incision. One portion of liver was frozen in
acetone and dry ice for later determination of glycogen.
Another portion of liver and the diaphragm sample were
sliced, and 90- to 110-mg pieces were incubated in a bicarbon-
ate-buffered Krebs-Ringer medium containing 5 mM glucose,
0.5 mM palmitate, 2% BSA, and either 0.3 or 0.6 µCi of
[1-14C]palmitate (for liver and diaphragm tissue, respec-
tively). After 2 h, 0.2 ml of hyamine was added to the center
wells of the incubation flasks (to trap CO2 ), and cellular Fig. 1. Effects of 1, 5, and 10 mg/kg methyl palmoxirate (MP) on ad
reactions were terminated by addition of 0.5 ml of 1 N H2SO4 libitum food intake in rats. Values are means ⫾ SE; n ⫽ 10
to the media. CO2 was collected for 30 min. Total lipids were rats/group. * P ⬍ 0.05 vs. vehicle treatment.
obtained by extraction of the tissue slices with heptane. Acid
water soluble products (AWSP) were extracted from the
media with chloroform:methanol:acid NaCl. Incorporation of
radioactivity into CO2, AWSP, and total lipids was measured intakes were measured 3 h later. The feeding test was
using a liquid-scintillation spectrometer. started at 5 h posttreatment because the first experi-
Data analysis. Dose-response data from experiments involv- ment showed that the eating response to the higher
ing more than two dose levels were analyzed by ANOVA. Time doses of the inhibitor is initiated at about this time.
course data were analyzed by ANOVA with repeated mea- As shown in Figs. 2 and 3, MP altered food intake and
sures. All other data involving two-group comparisons were plasma glucose and FFA concentrations in a dose-
analyzed using the Student’s t-test. Post hoc comparisons,
related fashion, but its effect on plasma ketone body
using a critical difference test or Newman-Keuls, were made
only when the main or interaction terms of the ANOVA were levels was not dose related. MP decreased plasma
statistically significant at P ⬍ 0.05. Only the results of post glucose and increased plasma FFA [F(6,72) ⫽ 10.8 and
hoc comparisons with a P ⬍ 0.05 are reported. 11.9 for glucose and FFA, respectively, P ⬍ 0.00001].
Plasma glucose and FFA levels of rats given vehicle or 1
RESULTS mg/kg MP were similar and did not change signifi-
cantly from baseline (0 h) over the 5-h observation
Ad libitum food intake. To determine the dose- period. However, 3 and 5 h after administration of 5 or
response relationship between MP and feeding behav- 10 mg/kg MP, plasma glucose levels decreased and FFA
ior, 40 rats (n ⫽ 10/group) were given either vehicle or concentrations increased relative to treatment with
1, 5, or 10 mg/kg MP, and food intakes were measured vehicle or 1 mg/kg MP and relative to baseline values.
every 2 h for 8 h. Administration of MP increased food The effect of 5 mg/kg MP on glucose and FFA levels was
intake of rats in a dose-related fashion during the 8-h similar to that of 10 mg/kg MP. MP treatment de-
observation period [Fig. 1; F(12,144) ⫽ 4.3, P ⬍ .0001]. creased plasma ketone body levels [F(6,72) ⫽ 19.0, P ⬍
Compared with those given vehicle or 1 mg/kg MP, rats 0.00001]. In rats given vehicle, plasma ketone body
given the two larger doses of MP increased food intake concentrations increased significantly from baseline
starting, respectively, 4 and 6 h after MP treatment (0 h) during the 5-h period after treatment. In MP-
(P ⬍ 0.05). Food intakes of rats given the lowest dose (1 treated rats, ketone body levels decreased markedly
mg/kg) were similar to those of rats given vehicle. from baseline during this period and were significantly
Plasma fuels and food intake. The effects of MP on lower than those in vehicle-treated rats at both the 3-
circulating metabolic fuels and food intake were as- and 5-h time points. Administration of MP had no
sessed by sampling plasma fuels just before the feeding significant effect on plasma concentrations of glycerol
test, a protocol that circumvents the confounding effect or triglycerides (data not shown).
of food ingestion on these metabolic measures (e.g., When rats were refed after the last blood sample,
Refs. 5–7). One week after the first experiment, the those given MP increased food intake in a dose-related
same rats were deprived of food, and tail blood was fashion [F(3,36) ⫽ 10.0, P ⬍ 0.0001; Fig. 3]. Whereas
sampled just before the animals were retreated with rats given 1 mg/kg MP ate the same amount of food as
vehicle or MP. Additional blood samples were collected those given vehicle, rats given the higher doses of MP
3 and 5 h later. Food was returned (at 5 h), and food consumed more food than did rats treated with vehicle

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R1048 FATTY ACID OXIDATION AND FOOD INTAKE
Fig. 2. Effects of 1, 5, and 10 mg/kg MP on plasma fuels in rats.
Values are means ⫾ SE; n ⫽ 10 rats/group. * Significantly different
from vehicle and 1 mg/kg MP (P ⬍ 0.05); † significantly different from
vehicle treatment (P ⬍ 0.05).
or 1 mg/kg. In addition, rats given 10 mg/kg MP ate
more than those treated with 5 mg/kg.
Plasma glucose and food intake. The previous experi-
ment showed that doses of MP that decreased plasma
glucose concentration also stimulated food intake. Be-
cause a reduction in plasma glucose is thought to be a
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signal that triggers feeding behavior (e.g., Ref. 30), we
performed an experiment to determine whether prevent-
ing the decline in plasma glucose level after MP treat-
ment would attenuate the eating response to the inhibi-
tor. Twenty-one rats were deprived of food and given
vehicle. Five hours later, blood samples were taken
from the tips of their tails (for analysis of glucose), food
was returned, and intakes were measured 3 h later.
Two or three days later, rats were retested, except that
they were given 10 mg/kg of MP instead of vehicle and
subcutaneously injected 4 h later with either 0.5 ml
isotonic saline or 25% glucose to raise systemic glucose
levels.
Under control (vehicle) conditions, plasma glucose
concentrations ranged from 6.5 to 7.6 mM. Although
rats injected with glucose had higher plasma glucose
levels 5 h posttreatment with MP than did those
injected with saline (6.7 ⫾ 0.1 vs. 6.3 ⫾ 0.2 mM), this
difference was not statistically significant, because
glucose values were somewhat variable in the two
groups. To assess the relationship between plasma
glucose level and the eating response to MP treatment,
we compared the results from glucose-injected rats that
had plasma glucose concentrations ⱖ6.5 mM (n ⫽ 7)
with those from saline-injected rats that had glucose
levels ⬍6.5 mM (n ⫽ 7). A plasma glucose concentration
of 6.5 mM was chosen as the criterion because this was
the lowest level found in vehicle-treated rats. As shown
in Table 1, rats injected with glucose had significantly
greater plasma glucose concentrations than did those
given saline [t(12) ⫽ 4.7, P ⬍ 0.0005] and showed less of
a decrease in plasma glucose from control (vehicle)
Fig. 3. Effects of MP on food intake in rats. Access to food was delayed
for 5 h after treatment, and intakes were measured after 3 h of
refeeding. Values are means ⫾ SE; n ⫽ 10 rats/group. V, vehicle.
Values labeled with different letters are significantly different (P ⬍
0.05).
 FATTY ACID OXIDATION AND FOOD INTAKE R1049

Table 1. Effect of exogenous glucose on plasma glucose 0.001], but only the high dose was effective, decreasing
and food intake of rats treated with MP incorporation by 32%. There was no significant effect of
MP treatment on incorporation of radiolabel into AWSP
Plasma ⌬Plasma Food ⌬Food or total lipids in diaphragm.
Glucose, Glucose, Intake, Intake,
n mM mM g/3 h g/3 h Hepatic energy status. The results of the previous
experiment showed that different doses of MP, which
Saline 7 6.08 ⫾ 0.13 ⫺0.93 ⫾ 0.07 1.4 ⫾ 0.4 1.0 ⫾ 0.3
Glucose 7 6.99 ⫾ 0.14* ⫺0.24 ⫾ 0.17* 1.9 ⫾ 0.4 1.2 ⫾ 0.4
are differentially effective in stimulating food intake,
decreased hepatic fatty acid oxidation to the same
Values are means ⫾ SE; n ⫽ no. of rats. Rats were given 10 mg/kg extent. Fatty acids are an important hepatic fuel (14,
methyl palmoxirate (MP) by gavage in methyl cellulose vehicle and
deprived of food. Four hours later, they were injected subcutaneously
15), and recent evidence suggests that a decline in liver
with 0.5 ml of either isotonic saline or 25% glucose. One hour later, ATP content generates a signal that triggers eating
tail blood samples were taken and food was returned, and intakes behavior (3, 11). Therefore, to determine whether doses
were measured 3 h later. Values for ⌬plasma glucose and ⌬food of MP that differentially affect food intake also have
intake are differences from baseline (vehicle treatment). * Signifi- different effects on liver energy status, we measured
cantly different from saline (P ⬍ 0.05).
ATP, ADP, and Pi in 24 rats (n ⫽ 8/group) 5 h after
administration of vehicle or 1 or 10 mg/kg MP. As in the
levels [t(12) ⫽ 3.7, P ⬍ 0.005] than did those injected previous experiments, rats were deprived of food after
with saline. Despite these differences in plasma glu- treatment to avoid confounding the metabolic mea-
cose, food intakes after MP treatment in saline- and sures with consequences of food ingestion.
glucose-injected rats were similar considered either in Administration of the high, but not low, dose of MP
absolute terms or as a change from control (vehicle) reduced hepatic energy status (Fig. 6). Overall, MP
levels. Rats injected with saline (n ⫽ 3) and glucose decreased liver ATP content and reduced the hepatic
(n ⫽ 4) that did not meet the criterion described above ATP-to-ADP ratio and phosphorylation potential
did not differ significantly with respect to plasma [F(2,21) ⫽ 5.3, 13.2, and 9.1 and P ⬍ 0.02, 0.001, and
glucose level, change in plasma glucose (from vehicle- 0.002, respectively]. These effects were attributable to
treated control condition), or eating response to MP. the high dose of MP. Only the 10-mg/kg dose signifi-
Tissue FFA metabolism. Given the observation that cantly reduced liver ATP relative to vehicle treatment.
all doses of MP decreased plasma ketone bodies, but Also, the hepatic ATP-to-ADP ratio and phosphoryla-
only the high doses affected plasma glucose and fatty tion potential were lower only in rats given 10 mg/kg
acids, it appeared that the lowest dose of MP had a MP compared with those given vehicle or 1 mg/kg. Rats
more selective effect on liver fatty acid oxidation than given the low dose did not differ from those given
did the higher doses. To determine whether the inhibi- vehicle on any of these measures. MP treatment also
tion of fatty acid oxidation induced by the low dose was
relatively restricted to liver, we examined the effects of
low and high doses of MP on fatty acid metabolism in
liver and muscle in vitro. To relate these metabolic
measures to the effects of MP on food intake, samples of
liver and muscle tissue were collected under conditions
in which different doses of MP were shown to differen-
tially affect feeding behavior. Thirty rats (n ⫽ 10/group)
were deprived of food and given either vehicle or 1 or 10
mg/kg MP. Five hours later (by which time rats initiate
eating in response to the high dose), rats were killed for
collection of liver and diaphragm tissue and measure-
ment of liver glycogen and in vitro FFA metabolism.
MP treatment reduced liver glycogen content
[F(2,27) ⫽ 32.6, P ⬍ 0.00001; Fig. 4]. Both doses of MP
significantly reduced liver glycogen, and the high dose
decreased glycogen significantly more than did the low
dose (78 vs. 40%).
Administration of MP reduced the incorporation of
14C (from [1-14C]palmitate) in liver tissue into CO and
2
AWSP and increased the incorporation into total lipids
[F(2,27) ⫽ 5.9, 26.6, and 3.8 and P ⬍ 0.01, 0.00001, and
0.05 for CO2, AWSP, and total lipids, respectively; Fig.
5]. The effects of the two doses of MP on incorporation of
radioactivity into CO2 and AWSP were similar (a de-
crease of ⬃25 and 55%, respectively), but only the high
Fig. 4. Effects of MP on liver glycogen in rats. Rats were deprived of
dose had a significant effect on incorporation into total food on treatment, and liver samples were collected 5 h later. Values
lipids. In muscle (diaphragm) tissue, MP significantly are means ⫾ SE; n ⫽ 10 rats/group. Values labeled with different
reduced 14C incorporation into CO2 [F(2,27) ⫽ 10.8, P ⬍ letters are significantly different (P ⬍ 0.05).

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R1050 FATTY ACID OXIDATION AND FOOD INTAKE

tion to the same extent as did the low dose but in

addition suppressed fatty acid oxidation in muscle.
Whereas the eating response to MP treatment could
not be explained by changes in hepatic fatty acid
oxidation, the behavioral effect of MP was found to
track reductions in liver glycogen content and hepatic
energy status. These findings indicate that a decrease
in hepatic fatty acid oxidation per se may not be
sufficient to trigger eating behavior. Instead, the re-
sults suggest that, when combined with a limited
capacity to oxidize alternative fuels, a decrease in
hepatic fatty acid oxidation can stimulate feeding behav-
ior by limiting hepatic energy production.
Metabolic effects of MP treatment. The metabolic
effects of MP treatment observed in these experiments
are consistent with the inhibitor’s actions that have
been described previously (5, 7, 18, 31–35). Although
high doses of MP inhibit CPT I in liver and muscle, low
doses, like the 1 mg/kg dose used in the present
experiments, act primarily on CPT I in liver (32),
possibly because MP has a greater affinity for the liver
isoform of CPT I than it does for the muscle isoform
(12). This selective effect of low doses of MP accounts for
the observation that, whereas both low and high doses
inhibited fatty acid oxidation in liver tissue, only the
high dose suppressed fatty acid oxidation in dia-
phragm. In keeping with the greater susceptibility of
liver CPT I to inhibition by MP, low and high doses of
the inhibitor suppressed hepatic fatty acid oxidation
maximally. As a consequence, the different doses also

Fig. 5. Effects of MP on in vitro fatty acid metabolism in liver and

diaphragm slices. Rats were deprived of food on treatment, and tissue
samples were collected 5 h later. Values are rates of incorporation of
14C (from [1-14C]palmitate) into CO (top), acid water soluble prod-
2
ucts (AWSP; middle), and total lipids (bottom). Values are means ⫾
SE; n ⫽ 10 rats/group. Values labeled with different letters are
significantly different (P ⬍ 0.05).

reduced liver glycogen content [F(2,21) ⫽ 34.2, P ⬍

0.00001]. As in the last experiment, both doses of MP
depleted liver glycogen, and the high dose reduced it
more than twice as much as did the low dose (71 vs.
27%).
DISCUSSION

These experiments were conducted to determine

whether a decrease in fatty acid oxidation restricted to
the liver stimulates feeding behavior in rats. In vivo
and in vitro metabolic measures verified that a low dose
of MP (1 mg/kg) selectively decreased fatty acid oxida-
tion in the liver. Despite this inhibition, MP given at
Fig. 6. Effects of MP on liver ATP, ATP/ADP (top), phosphorylation
this dose did not stimulate food intake. In contrast, potential, and glycogen (bottom). Values are means ⫾ SE; n ⫽ 10
feeding behavior increased after rats were given higher rats/group. Values labeled with different letters are significantly
doses of MP, which inhibited hepatic fatty acid oxida- different (P ⬍ 0.05).

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 FATTY ACID OXIDATION AND FOOD INTAKE
produced an equivalent and maximal decrease in liver
ketogenesis, judging by the similar declines in plasma
ketone body levels. Given the differences in the site of
action of low and high doses of MP, the increase in
plasma FFA in rats given the high, but not low, doses of
MP appears to result from a decrease in the uptake
and/or utilization of fatty acids primarily by muscle as
opposed to liver.
Many of the metabolic effects of MP can be under-
stood in terms of the glucose-fatty acid cycle described
by Randle et al. (17), in which a decrease in the
oxidation of fatty acids is met by a compensatory
increase in the oxidation of glucose and vice versa. This
reciprocal relationship between the metabolism of fatty
acids and glucose has been demonstrated in muscle, in
which glucose oxidation increases markedly after treat-
ment with MP (31, 35). The decrease in blood glucose
after administration of MP results in part from the
enhanced rate of glucose oxidation in muscle and also
in part from suppression of hepatic gluconeogenesis,
which is fueled largely by the oxidation of fatty acids
(33). As a result of the reduction in gluconeogenesis,
liver glycogen must be mobilized to meet the increased
demand of muscle tissue for glucose, and liver glycogen
content decreases accordingly. In the present experi-
ments, the decline in plasma glucose after MP treat-
ment was seen only with the higher doses of the
inhibitor, because these doses, which depressed muscle
fatty acid oxidation, probably also increased muscle
glucose oxidation. Presumably, the low dose of MP did
not decrease plasma glucose concentrations because it
failed to inhibit muscle fatty acid oxidation and produce
a compensatory change in glucose utilization. The
extraordinary depletion of liver glycogen in rats given
the high dose of MP most likely can be attributed to
enhanced glucose utilization by muscle. However, such
a mechanism does not explain the decrease, albeit
smaller, in liver glycogen in rats given the low dose of
MP because there was no evidence that muscle was
affected at this dose (see also Ref. 32). It is possible that
the reduction in liver glycogen in rats given the low
dose of MP resulted from an enhanced rate of glucose
oxidation by liver. Such an increase in hepatic glucose
oxidation would help explain how liver energy produc-
tion was maintained in rats given the low dose of MP
despite the inhibition of hepatic fatty acid oxidation.
Relationship between the metabolic and behavioral
effects of MP. Scharrer and colleagues (25) have sug-
gested that changes in hepatic fatty acid oxidation
produce a signal that controls food intake. They hypoth-
esize that this signal is generated when ketone bodies,
which are synthesized by the hepatocellular oxidation
of fatty acids, are released into the extracellular com-
partment and stimulate vagal afferent neurons inner-
vating the liver (25). The present results contradict this
hypothesis because feeding behavior after MP treat-
ment was not tied to either hepatic fatty acid oxidation
or to ketogenesis. Administration of 1 or 10 mg/kg MP
to rats induced a marked and equivalent inhibition of
hepatic fatty acid oxidation, but only the high dose
increased food intake. Inhibition of fatty acid oxidation
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R1051
was demonstrated under conditions similar to those
used in the behavioral tests and within a time frame in
which the behavioral responses were observed. Suppres-
sion of fatty acid oxidation was evidenced by a large
decline in circulating ketone bodies and a reduced
incorporation of 14C (from [1-14C]palmitate) into CO2
and AWSP in liver slices. Under the conditions used in
the in vitro experiments with liver tissue, it seems
likely that 14C recovered in AWSP includes ketone
bodies synthesized from the [1-14C]palmitate substrate.
That both low and high doses of MP decreased plasma
ketone body levels and apparently suppressed hepatic
ketogenesis to the same degree argues against the
notion that blood or liver ketone body concentrations
are monitored by the nervous system to elicit eating
behavior. However, because injection of ketone bodies
transiently decreases food intake in rats (e.g., Ref. 13),
it remains possible that the level of ketone body or some
aspect of their hepatic metabolism may play a role in
satiety.
Other metabolic effects of MP besides hepatic fatty
acid oxidation varied as a function of dose in the same
manner as did food intake. Administration of high
doses of MP decreased plasma glucose concentrations,
whereas treatment with the low dose had no such
effect. Decreased blood glucose has long been thought to
stimulate eating (e.g., Ref. 30). However, in the present
case, the decline in plasma glucose after the high dose
did not appear to underlie the eating response, because
injections of glucose that increased plasma glucose
levels did not prevent the increase in food intake after
MP treatment. This conclusion is consistent with other
results showing that MP treatment potentiates the
eating response to administration of other metabolic
inhibitors independently of changes in plasma glucose
levels (7, 18). In addition, the decline in plasma glucose
after MP treatment in the present experiment was
relatively small, leaving circulating levels well above
those needed to elicit eating behavior during insulin-
induced hypoglycemia (i.e., below 4 mM plasma) (30).
MP treatment decreased liver glycogen content, and
the magnitude of this depletion depended on the dose of
MP; the high dose decreased glycogen content at least
twice as much as did the low dose. Several investigators
have suggested that a reduction in liver glycogen
content generates a signal that stimulates food intake
(2, 22). In general, the present findings are consistent
with this hypothesis because the high dose of MP,
which produced the greatest depletion of liver glycogen,
increased feeding behavior, whereas the low dose did
not. However, the low dose of MP had a substantial
effect on liver glycogen (⬃30–40% decrease) yet had no
effect on food intake. In addition, previous experiments
have shown that administration of MP to rats synergis-
tically increases the eating response to injection of
2,5-anhydro-D-mannitol (2,5-AM), which inhibits glyco-
genolysis and prevents the depletion of liver glycogen
otherwise produced by MP (18). These and other find-
ings (7) argue strongly against a causal role of glycogen
depletion in the eating response to MP treatment.
R1052 FATTY ACID OXIDATION AND FOOD INTAKE
Of the metabolic parameters examined, changes in
hepatic energy status best paralleled the behavioral
responses to different doses of MP. Administration of
the high dose of MP decreased liver ATP concentration,
the hepatic ATP-to-ADP ratio, and phosphorylation
potential, whereas the low dose had no effect on these
measures. These changes in ATP, ADP, and Pi caused by
treatment with the high dose of MP are indicative of a
decrease in ATP production. Studies with 2,5-AM and
other metabolic inhibitors suggest that a reduction in
liver ATP or a related change in ATP metabolism
triggers eating behavior in rats (3, 11). The present
experiments are the first to show that administration of
an inhibitor of fatty acid oxidation that stimulates
feeding behavior also decreases liver energy produc-
tion, and thus provide additional evidence that changes
in hepatic energy status control food intake. The eating
response to injection of 2,5-AM is associated with a
larger decrease in ATP (11, 18) than that observed after
MP treatment in the present study. Whether this
reflects the operation of different signaling mecha-
nisms or the more acute action of 2,5-AM compared
with MP remains to be determined.
Perspectives
The results suggest that no single change in liver
metabolism at the substrate level generates a signal to
initiate food intake. Instead, the findings indicate that
a combined decrease in the oxidation of different fuels
can lead to a reduction in hepatic energy status, which
in turn triggers eating behavior (see Ref. 3). The liver
relies in large part on the oxidation of fatty acids for its
energy needs (14, 15); however, when the capacity to
oxidize fatty acids is limited, the liver must use other
fuels to maintain energy status. In addition to amino
acids (15), glucose derived from the breakdown of liver
glycogen could be used as an alternative fuel. However,
when both fatty acid oxidation is insufficient and the
supply of alternative fuels is limited or inaccessible,
hepatic energy production declines. It is under these
circumstances that a hepatic signal to eat is generated.
The current findings are consistent with this sce-
nario, although it is possible that under other condi-
tions in which fatty acid oxidation is suppressed differ-
ent mechanisms may be involved. In the present studies,
rats had a relatively low liver glycogen content to begin
with because they were fed a high-fat, low-carbohy-
drate diet (e.g., Ref. 18). Treatment with the high dose
of MP largely depleted this small store of liver glycogen,
presumably by increasing glucose demand from muscle.
According to the model described above, the suppres-
sion of liver fatty acid oxidation combined with a
depleted liver glucose reserve led to a decrease in
hepatic energy production and an eating response. In
comparison with the high dose, administration of the
low dose of MP suppressed liver fatty acid oxidation to
the same extent but did not deplete hepatic glycogen
reserves nearly as much. This differential effect on liver
glycogen content presumably resulted because the low
dose of MP did not suppress muscle fatty acid oxidation
and consequently did not produce a compensatory
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increase in muscle glucose oxidation. The relatively
larger glycogen reserve of rats given the low dose of MP
supplied fuel (glucose) for the liver, which, when oxi-
dized, forestalled a decline in hepatic energy produc-
tion and the generation of a signal to initiate feeding
behavior. In this way, the response of rats given 1 mg/kg
MP resembles that of rats fed a low-fat, high-carbohy-
drate diet and given 10 mg/kg MP, namely, a relatively
small decline in liver glycogen (7), in this case from a
high level, and no increase in food intake (7, 8).
For the model outlined here, a decrease in hepatic
fatty acid oxidation elicits feeding behavior when alter-
native fuels are not sufficiently available and/or oxidiz-
able to maintain energy production in the liver. In
many cases, glycogen would provide the alternative
source of hepatic fuel in the form of glucose, although
other oxidizable substrates (e.g., lactate or amino acids)
might serve a similar role. Circulating glucose seems
unlikely to be used as a hepatic fuel under these
conditions because liver metabolism is geared toward
glucose output rather than uptake in the face of in-
creased glucose utilization by muscle.
It is important to emphasize that, in terms of the
perspective outlined here, it is the utilization of glyco-
gen as a fuel source, rather than the absolute level of
liver glycogen, that is crucial. This distinction between
access to the hepatic glycogen reserve and the size of
that reserve is illustrated clearly by the finding that
administration of 2,5-AM potentiates the eating re-
sponse to MP while at the same time preventing the
depletion of glycogen (18). A role for liver glycogen in
the control of food intake has been suspected for many
years (2, 22), but the relationship between this meta-
bolic parameter and feeding behavior is inconsistent (6,
18, 26, 28). From the analysis presented here, this
inconsistency could be explained by the use of an
inappropriate measure of liver glycogen (i.e., size of the
reserve), as well as by the indirect contribution that the
glycogen reserve makes in maintaining hepatic energy
production.
Under some circumstances, an increase in the oxida-
tion of fatty acids by liver could maintain hepatic
energy production and forestall eating behavior when
the availability and/or oxidation of other substrates
such as glycogen is limited. The normalization of food
intake seen when hyperphagic diabetic rats are fed a
high-fat diet may be a case in point. These animals
have extremely low glycogen levels (e.g., Ref. 6), display
very high rates of fat oxidation (4, 6), and, unlike their
counterparts fed a low-fat diet, have normal liver ATP
levels (9). In the context of the model presented here, it
is important to emphasize that for hepatic fatty acid
oxidation to prevent eating, it must proceed at a rate
that supports the energy status of the liver. Feeding
behavior would still be triggered if the increase in fatty
acid oxidation were not of sufficient magnitude. Such a
situation may obtain during fasting, perhaps the most
fundamental condition that stimulates feeding behav-
ior. In the absence of food, liver glycogen is rapidly
depleted, and whereas fatty acid oxidation increases, it
 FATTY ACID OXIDATION AND FOOD INTAKE
does not increase enough to prevent a decline in liver
ATP production (29).
The authors thank Mary Pollice for technical assistance.
This research was supported by National Institute of Diabetes and
Digestive and Kidney Diseases Grants DK-36339 and DK-53109.
Present address: R. B. Harris, Pennington Biomedical Research
Center, Louisiana State Univ. 6400 Perkins Rd., Baton Rouge, LA
70808.
Address for reprint requests and other correspondence: M. I.
Friedman, Monell Chemical Senses Center, 3500 Market St., Philadel-
phia, PA 19104–3308 (E-mail: friedman@monell.org).
Received 21 September 1998; accepted in final form 21 December
1998.
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