Carbon Tetrachloride–Induced Cirrhosis in Rats: Influence
of the Acute Effects of the Toxin on Glucose Metabolism
FRANÇOIS MION, ALAIN GÉLOËN, ELISABETH AGOSTO,
In animal models, conflicting results on the effect of
cirrhosis on glucose metabolism have been reported.
The use of various toxins as well as differences in experi-
mental protocols may be responsible for these contro-
versial data. However, differences may also be explained
by the fact that glucose metabolism has been evaluated
following different time intervals after cessation of the
toxic injury. Therefore, we have performed intravenous
glucose tolerance tests, euglycemic hyperinsulinemic
clamps (at 2, 6, and 30 mU/kgrmin insulin infusion rates),
and determination of peripheral tissue glucose meta-
bolic index (by [3H]2-deoxy-glucose injection) in rats
treated for 10 weeks with carbon tetrachloride, either 3
days (acute group) or 2 weeks (delayed group) after the
last CCl4 dose was administered. Cirrhosis was con-
firmed by liver histological analysis, and by a 22% (P
õ .05) decrease in 13C-aminopyrine demethylation. In the
acute group, whole-body glucose disposal was decreased
at the highest insulin infusion rate only (19.7 { 1.2 vs.
23.4 { 1.2 mg/kgrmin in controls, P õ .05). In contrast,
results of the delayed group were not different from con-
trols at any insulin infusion rate. Peripheral tissue glu-
cose metabolic index was significantly decreased in all
muscles tested in the acute group compared with con-
trols. A significant decrease of glucose utilization was
found in some but not all muscles in the delayed group
but was less pronounced than in the acute group. In
conclusion, this study showed that insulin sensitivity in
cirrhotic rats is time-dependent with regard to the last
CCl4 administration. These results must be taken into
account when using this experimental model of liver cir-
rhosis. (HEPATOLOGY 1996;23:582-588.)
Glucose intolerance in liver disease was recognized
as far back as 1906, when Naunyn1 coined the term
‘‘hepatogenous diabetes.’’ In the clinical setting, glucose
homeostasis is often impaired; diabetes mellitus may
Abbreviations: ABT, aminopyrine breath test; GIR, glucose infusion rate;
WAT, white adipose tissue; IBAT, interscapular brown adipose tissue.
From the Laboratoire de Physiologie and Unité de Recherche Associée,
Centre National de Recherche Scientifique D1341, Université Claude Bernard
Lyon 1, 69373 Lyon cedex 08, France.
Received March 29, 1995; accepted October 3, 1995.
Supported by a research grant from Inbiomed International, 10bis rue A.
Lumière, 69008 Lyon, France, and from Fondation Mérieux, Lyon, France.
Address reprint requests to: François Mion, M.D., Laboratoire de Physiolo-
gie Lyon-Nord, 8, avenue Rockefeller 69373 Lyon cedex 08, France.
Copyright q 1996 by the American Association for the Study of Liver
Diseases.
0270-9139/96/2303-0025$3.00/0
582
5p0b$$0028 02-20-96 20:13:36 hepa
AND YVES MINAIRE
occur in as many as 10% to 15% of cirrhotic patients.2,3
Furthermore, the majority of nondiabetic cirrhotic indi-
viduals are characterized by hyperinsulinemia in the
fasting state4,5 and excessive insulin response to an
oral or intravenous glucose load.6,7 Insulin resistance
thus appears as the main feature of glucose metabolism
in human cirrhosis.8,9 It is well accepted that hepatic
insulin sensitivity is maintained in cirrhosis,7,10 and
that the main site of insulin resistance in human cir-
rhosis is skeletal muscle (decreased glycogen synthe-
sis), although other tissues such as the white adipose
tissue may also play a significant role in other diseases
and species. At the cellular level, insulin resistance
most likely involves a postreceptor defect,2,8 but the
exact nature of this defect remains unknown.
Although studies based on human subjects are likely
to give the most relevant information, animal models
of cirrhosis are still clearly needed to investigate in
more detail the mechanisms of impairment in glucose
metabolism. Although animal models of cirrhosis are
scarce and often unreliable,11 many attempts have been
made in the past to study metabolic alterations of cir-
rhosis using animal models obtained with various tox-
ins administered by many different routes and follow-
ing variable schedules.12-15 Many conflicting results
have been reported, depending on the toxin used to
induce cirrhosis, and the delay observed between the
end of induction treatment and the metabolic experi-
mentations. In recent years, the most popular model
has been the carbon tetrachloride–induced cirrhosis in
rats.14-16 Although this model has already been used
for metabolic studies,15,17,18 no attempt has been made
so far to clearly evaluate the influence of CCl4 toxicity
on glucose metabolism, and thus the relevance of this
model to the study of the mechanisms of insulin resis-
tance in human cirrhosis.
After a careful evaluation of morphologic and meta-
bolic criteria needed to confirm the presence of cirrhosis
after chronic phenobarbital and CCl4 treatment, we ex-
amined the following in this study: (1) glucose tolerance
by intravenous glucose tolerance tests; (2) whole-body
insulin sensitivity by the euglycemic hyperinsulinemic
clamp technique; and (3) insulin-stimulated glu-
cose uptake by peripheral tissues, by injection of
[3H]2-deoxyglucose during euglycemic-hyperinsulin-
emic clamps, to determine the possible sites of insulin
resistance (skeletal muscle and white and brown adi-
pose tissues).
WBS: Hepatology
HEPATOLOGY Vol. 23, No. 3, 1996
These tests were performed in rats with liver cirrho-
sis induced by 10 weeks of CCl4 treatment, either 3
days (acute group) or 2 weeks (delayed group) after the
last CCl4 administration.
MATERIALS AND METHODS
Animals. Male Sprague-Dawley rats (Iffa-Credo, L’Ar-
bresle, France), weighing 180 to 220 g, were housed in indi-
vidual cages kept at 217 to 257C under 12-hour dark/light
cycles, with free access to standard laboratory chow and tap
water. Animals received humane care in compliance with our
institution’s guidelines.
Cirrhosis was obtained after a 10-week treatment by ad-
ministration of phenobarbital in the drinking water (350 mg/
L) and weekly CCl4 gavages; the toxin was administered in
increasing doses based on individual body weight variations,
as described by Proctor and Chamtara.16 The weekly CCl4
schedule was chosen as the best treatment regimen based on
a previous study showing that hepatic lesions were maximal
for this dosage.19
Control animals did not receive phenobarbital or CCl4 .
They were matched with treated animals based on body
weight.
Metabolic experiments were performed in CCl4-treated
rats and controls. One set of cirrhotic rats (referred to as the
acute group) were studied 3 days after the last CCl4 dose,
when the hepatotoxic effects of CCl4 are still present19; the
second set of cirrhotic rats (referred to as the delayed group)
were studied 2 weeks after the administration of the last
CCl4 dose to minimize the acute effects of the treatment. The
presence of cirrhosis was confirmed in all treated animals
used in this study by measurement of the relative liver, tes-
tes, and spleen weights, used as markers of cirrhosis and
portal hypertension17; gross macroscopic appearance and his-
topathological examination of the liver obtained at the end
of experiments; and determination of serum aspartate amino-
transferase activity and total bilirubin concentration on a
Cobras automatic analyzer (Roche, Neuilly, Seine, France).
[13C]-Aminopyrine Breath Test. The test was used to moni-
tor the decrease of hepatic metabolic capacity induced by
the chronic CCl4 treatment.19,20 In treated animals, the 13C-
aminopyrine breath test (ABT) was performed at week 1, 3,
5, and 10. Four milligrams per kilogram of body weight of
[N,N-dimethyl-13C2]-aminopyrine (99 Atom% 13C; Tracer
Technologies, Somerville, MA) were injected intraperitone-
ally; the 13CO2 enrichment in breath samples was measured
by GC-isotope ratio mass spectrometry,21 and the increase
(related to basal values) of cumulative 13CO2 excretion during
30 minutes was calculated according to the trapezoidal rule
and expressed in atom per excess (APE 13Cr30 minutes). The
test was performed just before the next CCl4 administration
to avoid the inhibitory effect of CCl4 on aminopyrine demeth-
ylation.19 The ABT was also performed in control rats body-
weight–matched with rats treated with CCl4 for 1 week and
10 weeks, the control rats being administered phenobarbital
in the drinking water for 1 week before the test.
Intravenous Glucose Tolerance Tests. The animals were
fasted 16 hours before the intravenous glucose tolerance
tests. After intraperitoneally-administered pentobarbital an-
esthesia, two polyethylene catheters (EO 3403, Biotrol, Vil-
leron, France) were inserted, one into the right jugular vein,
and the other into the left carotid artery. The carotid catheter
was used for blood sampling and the jugular for intravenous
infusion. 0.5 mg/kg of body weight of glucose were injected,
and 125-mL blood samples were taken before glucose adminis-
5p0b$$0028 02-20-96 20:13:36 hepa
MION ET AL. 583
tration and 2, 5, 7, 10, 15, 30, and 45 minutes thereafter to
measure plasma glucose (Glucose Analyser II, Beckman) and
insulin concentrations by radioimmunoassay (CIS Biointer-
national, Gif/Yvette, France).
Euglycemic-Hyperinsulinemic Clamp Studies. They were
carried out according to the technique of Kraegen et al.22
Vascular catheters were inserted just before the study, under
pentobarbitone anesthesia. Rats were infused with human
neutral insulin (Actrapid HM, 40 U/mL, Novo Industries,
Denmark) while maintaining blood glucose at basal level (6
mmol/L). Insulin was continuously infused at either 2, 6, or
30 mU/kgrmin (in 0.9% saline and 1% bovine fatty acid–free
serum albumin), at 16.6 mL/min for 2 hours to achieve steady
plasma insulin concentrations. Blood samples of 25 mL were
obtained at 5-minute intervals to determine plasma glucose
levels and of 100 mL at 0, 60, and 120 minutes of the clamp
to measure plasma insulin concentrations. To maintain eu-
glycemia, glucose at different concentrations (5%-50%) was
infused at variable rates using a peristaltic roller pump, the
delivery of which was controlled after each clamp. The needed
glucose infusion rate (GIR) plateaued between 60 and 120
minutes (GIR60-120= in mg glucose/kgrmin) and was consid-
ered as a measure of the whole-body glucose utilization.
Peripheral Tissue Glucose Metabolic Index. Glucose up-
take by individual tissues was estimated as the ‘‘tissue glu-
cose metabolic index.’’22 Briefly, 45 minutes before the end of
the clamp, the glucose analogue 2-[1,2-3H]-deoxy-D-glucose
(32.5 mCi; Isotopchim; Ganagobie-Peyruis, France) and [U-
14
C]-saccharose (4 mCi; DuPont de Nemours, Les Ullis,
France) were administered intravenously together as a bolus,
in 250 mL of saline solution containing 0.25% albumin. Fifty-
microliter plasma samples were collected at 0, 2, 5, 7, 10, 15,
20, 30, and 45 minutes after the administration of the tracer
bolus for estimation of the plasma radioactivity disappear-
ance curve. At the end of the clamp (insulin infusion rate of
30 mU/kgrmin), rats were killed by cervical dislocation and
the following tissues rapidly removed: diaphragm, muscles
of the hindlimb (extensor digitorum longus, tibialis, soleus,
plantaris, and gastrocnemius), white adipose tissue (WAT)
(epididymal and retroperitoneal), and interscapular brown
adipose tissue (IBAT). These tissues were immediately
weighed and frozen until subsequent analysis. Tissue and
plasma samples were solubilized in 2 mL potassium hydrox-
ide (3%) at 657C in a shaking water bath for 2 hours. To
reduce quenching, 100 mL of hydrogen peroxide (30%) was
added to the vials which were kept at 657C for an additional
1 hour. Ten milliliters of scintillant (Insta-Gel; Packard, Mer-
iden, CT) was then added and double-isotope counting per-
formed in a scintillation counter (SL 3000 Intertechnic; Kon-
tron, St-Quentin, France). Quench corrections were applied
on external standard. The extracellular volume of each tissue
sample was measured using 14C-saccharose and was used
to calculate the intracellular [3H]2-deoxyglucose-6-phosphate
content (C*m(T) dpm/g at T Å 45 min), according to the method
of Hom et al.23 Glucose uptake (ng glucose/mgrmin) was then
calculated as the ‘‘tissue glucose metabolic index’’ (Rg)22:
Cm*(T)
Rg Å dt
*
T
(Cp*/Cp)
0
3
where C*p is plasma H-2DG (dpm/mL) and Cp is plasma glu-
cose (mg/mL).
Statistical Analyses. All experimental data in the text or
in figures are expressed as mean { SEM. The results between
groups were examined by one-way ANOVA, and a Fischer’s
WBS: Hepatology
584 MION ET AL. HEPATOLOGY March 1996

TABLE 1. Liver, Spleen, and Testes Weight (related to body

weight), Serum Bilirubin, and Aspartate Amino
Transferase Levels in Control, Acute Cirrhotic,
and Delayed Cirrhotic Rats
Acute Delayed
Controls Cirrhosis Cirrhosis

Body weight (g) 478 { 23 480 { 15 472 { 8

Liver (g/kg bw) 28.4 { 1.2 46.5 { 3.7*† 35.1 { 1.2*
Spleen (g/kg bw) 1.9 { 0.2 3.6 { 0.4* 3.1 { 0.2*
Testes (g/kg bw) 8.2 { 0.4 6.9 { 0.5* 6.8 { 0.2*
Bilirubin ( mmol/L) 3 { 1 35 { 7*† 2.5 { 1
AST (IU/L) 55 { 4 342 { 36*† 134 { 7*

NOTE. The results represent the mean { SEM of 7 to 23 sepa-

rate experiments.
Abbreviations: bw, body weight; AST, aspartate aminotransfer-
ase.
* Significant difference between controls and cirrhotic rats.
† Significant difference between the acute and delayed groups of
cirrhotic rats (P õ .01).

post-hoc test was used for group comparisons. Statistical sig-

nificance was set at P õ .05.
RESULTS
Evaluation of Cirrhosis in CCl4-Treated Animals.
Morphological and biological data of control and
treated rats are shown in Table 1. Cirrhosis was con-
firmed in all treated animals by histopathological anal-
ysis. Nodules of hepatic parenchyma were clearly delin-
eated by thick fibrous bands (Fig. 1). Ascites was absent
in all cirrhotic animals in the delayed group, whereas
it was present in some cirrhotic rats of the acute group.
Finally, the results of ABT performed after 10 weeks
of CCl4 treatment was significantly decreased when
compared with the initial ABT results performed in the
same animals at the beginning of treatment (022%)
FIG. 1. Histological section of the liver of a CCl4-treated rat, ob-
tained 2 weeks after the end of treatment. Nodules of hepatocytes
are separated by thick fibrous bands, defining the typical aspect of
cirrhosis. (Hematoxylin and eosin; original magnification 160.)
FIG. 2. Evolution of 13C-aminopyrine breath test during chronic
phenobarbital-CCl4 treatment to obtain cirrhosis. The breath test
was also performed in body weight–matched controls after 1 week
of phenobarbital treatment (n Å 10 for each point). *, Significant
difference in treated rats between one and 5 and 10 weeks of CCl4
treatment. †, Significant difference of the ABT between rats after 10
weeks of CCl4 treatment and body weight–matched controls.
and to the ABT of controls of similar body weight
(031%) (Fig. 2).
Glucose Tolerance Tests. After a 16-hour fast, basal
plasma glucose and insulin levels were not different in
the acute (n Å 6) and delayed (n Å 6) cirrhotic rats,
compared with controls (n Å 5) (plasma glucose, 6.1
{ 0.2 and 6.7 { 0.1 vs. 6.4 { 0.2 mmol/L; plasma insu-
lin concentration, 13.2 { 3.1 and 13.0 { 1.7 vs. 12.5
{ 0.8 mU/L, respectively). The plasma glucose re-
sponse curves after intravenous administration of 0.5
g glucose/kg BW were identical in cirrhotic rats of the
delayed group compared to controls (Fig. 3A). The
shape of the glucose response curve was similar in the
acute group; however, the area under the curve of the
incremental plasma glucose between 0 and 45 minutes
was decreased in the acute cirrhosis group compared
to the delayed group and controls (126.2 { 4.3 vs. 193.5
{ 6.5 and 213.7 { 5.8 mmol/Lr45 min, respectively; P
õ .05). The percentage of variations of plasma insulin
concentration over baseline was similar in the three
groups of rats (Fig. 3B).
Whole-Body Glucose Utilization During Euglycemic-
Hyperinsulinemic Clamps. Plasma insulin levels mea-
sured during the last hour of the clamp are summarized
in Fig. 4A (n Å 7 to 8 rats for each group and each
value of clamp). Plasma insulin concentrations were
greater in the acute cirrhosis group, reaching statisti-
cal significance at 2 and 30 mU/kgrmin insulin infu-

5p0b$$0028 02-20-96 20:13:36 hepa WBS: Hepatology

HEPATOLOGY Vol. 23, No. 3, 1996
FIG. 3. Evolution of (A) incremental plasma glucose levels and (B) percentage of variation of plasma insulin concentration over baseline
after an intravenous glucose load (0.5 g/kg body weight) in controls, and in the delayed and acute groups of cirrhotic rats.
sion rate. In the delayed group, plasma insulin levels
were significantly greater than in the control group at
30 mU/kgrmin, but not different at physiological insu-
lin infusion rates (2 and 6 mU/kgrmin).
Glucose infusion rates (GIR60-120) needed to maintain
basal plasma glucose concentration (6 mmol/L) are pre-
sented in Fig. 4B. In the acute group, the GIR60-120 was
significantly greater than in the control group at 2 mU/
kgrmin insulin infusion rate, and lower at 30 mU/
kgrmin (P õ .05). In contrast, no difference was found
between the delayed and the control groups.
Thus, in the range of physiological plasma insulin
concentrations (20-100 mU/L), insulin sensitivity was
identical in the three groups of rats; the higher
GIR60-120 found in the acute group at 2 mU/kgrmin
insulin infusion rate is explained by higher plasma in-
sulin concentrations. At supraphysiological doses of in-
sulin infusion (30 mU/kgrmin), insulin resistance was
evident only in the acute group.
Glucose Metabolic Index in Individual Tissues. The
glucose metabolic index (Rg) was considered as repre-
sentative of insulin-stimulated glucose uptake in differ-
ent types of tissues (muscles and adipose tissue). The
results of glucose uptake in these peripheral tissues
obtained at 30 mU/kgrmin insulin infusion rate for
controls and cirrhotic rats (acute, n Å 7; delayed
groups, n Å 8) are shown in Fig. 5. The Rg in epididymal
and retroperitoneal WAT was identical in the three
groups of rats. In IBAT, the Rg was similarly decreased
in the acute and delayed cirrhosis groups compared
with controls (P õ .05). With regard to muscles, the
diaphragm, the only working muscle under our experi-
mental conditions, showed the highest Rg , whereas glu-
cose uptake was in the same range for all resting mus-
cles. There was a significant decrease of glucose uptake
in all muscles tested in the acute group compared with
controls. In the delayed cirrhosis group, Rg was signifi-
cantly lower in diaphragm, tibialis, and gastrocnemius;
in the other muscles tested, there was a tendency to-
5p0b$$0028 02-20-96 20:13:36
MION ET AL. 585
ward decreased Rg versus controls, although this differ-
ence did not reach statistical significance. Finally, Rg
was significantly lower in diaphragm and tibialis in
the acute versus delayed group; Rg also tended to be
lower in the other muscles of the former group.
DISCUSSION
In our study, chronic oral phenobarbital-intragastric
CCl4 treatment produced a typical liver cirrhosis as
described by Proctor and Chamtara.16 The cirrhosis
was confirmed by morphological and histological data
obtained at the end of experiments for each rat. The
decrease of ABT results at the end of CCl4 treatment
was also confirmative of the efficacy of this protocol to
obtain a significant reduction of the liver functional
mass.19,20
Thus, this animal model might allow the study of the
mechanisms of cirrhosis-induced alterations of glucose
metabolism. However, CCl4 , besides its well-known
hepatotoxicity, also has acute toxic effects on many or-
gans, including kidneys24 and pancreas,12,25 that could
interfere with the metabolic effects of cirrhosis. Fur-
thermore, the time dependency of CCl4 acute hepato-
toxicity is well known,19 and the results of metabolic
studies could vary depending on when these are per-
formed after the last CCl4 administration. To study a
possible interference of the acute effects of the toxin,
metabolic studies were performed in CCl4-adminis-
tered cirrhotic rats at two different schedules after the
end of treatment. In one group of rats (acute group),
metabolic experiments were performed only 3 days
after the last CCl4 administration; in this situation,
rats are clearly ill, body weight loss is maximal at that
time (around 10%), ascites may be present, and liver
functions are still severely depressed because of direct
CCl4 hepatotoxicity.17,19 In a second group of cirrhotic
rats (delayed group), experiments were performed 2
weeks after the end of treatment; rats have regained
some weight at that time, and CCl4 and its toxic metab-
hepa WBS: Hepatology
586 MION ET AL.
olites have been cleared out of the organism.26 The
choice of control rats is difficult with this chronic treat-
ment model; using sham-treated animals would have
given a control group with higher body mass, a situa-
tion that by itself could produce significant differences
in glucose metabolism.13 Thus, we decided to use as
controls untreated animals matched for body weight
with CCl4-treated rats.
Our results showed that neither acute nor delayed
FIG. 4. Plasma insulin levels (upper panel) obtained during the
last hour of the euglycemic hyperinsulinemic clamps in control rats
and in the acute and delayed groups of cirrhotic rats. In the same
groups of rats, GIR needed to maintain steady euglycemia (6 mmol/
L) between 60 and 120 minutes after the beginning of euglycemic
hyperinsulinemic clamp studies. *, Indicates a significant difference
versus controls; 7, between the two groups of cirrhotic rats.
5p0b$$0028 02-20-96 20:13:36 hepa
HEPATOLOGY March 1996
FIG. 5. Variations of the glucose metabolic index (Rg , represent-
ing glucose uptake) in different muscles, white and brown adipose
tissues, in controls, acute and delayed groups of cirrhotic rats, at 30
mU/kgrmin insulin infusion rate. The vertical bars represent the
mean of seven to eight separate experiments for each group. *, Indi-
cates a significant difference between controls and cirrhotic rats. †,
Indicates a significant difference between cirrhotic rats in the acute
group versus the delayed group.
cirrhotic rats were glucose intolerant. Basal plasma
glucose insulin concentrations were identical in con-
trols and treated rats (acute and delayed), in accor-
dance with previous reports.18,27 In contrast, Naka-
mura et al.12 found a decreased insulin content in the
pancreas of CCl4-treated cirrhotic rats, whereas ele-
vated plasma levels of insulin and glucagon were ob-
served in studies of thioacetamide-induced rat cirrho-
sis.25,28 These variations may be partially caused by the
toxin used to induce cirrhosis rather than by cirrhosis
itself; CCl4 and thioacetamide have been shown to in-
duce profound changes in relative pancreatic weight
and hormonal contents.12,25 In vitro basal and stimu-
lated releases of insulin from isolated pancreatic islets
were found to be decreased in CCl4-treated cirrhotic
rats,28 a finding that could explain the discrepancy be-
tween basal plasma insulin concentrations in rat and
human cirrhosis. Another interesting finding of the in-
travenous glucose tolerance test studies was the de-
creased plasma glucose response curve found in the
acute group, compared with the delayed group and con-
trols, without any significant difference in insulin re-
sponse. This may relate to the toxic effects of CCl4 ;
when tested 3 days after CCl4 administration, rats are
still severely ill and have been fasting for most of this
period. Glycogen contents are extremely low in most
tissues,17 and thus the injected glucose must be avidly
taken by peripheral tissues, explaining the decreased
hyperglycemic response in this group.
Euglycemic-hyperinsulinemic clamp studies were
performed to measure insulin resistance in cirrhotic
rats. At physiological plasma insulin concentrations
(20-100 mU/L), the only difference in GIR between the
three groups was an increase in the acute group at 2
WBS: Hepatology
HEPATOLOGY Vol. 23, No. 3, 1996
mU/kgrmin insulin infusion rate, probably related to
the higher plasma insulin levels obtained in this group.
The increased plasma insulin concentration in the
acute group may be caused by the profound inhibition
of liver metabolism induced by CCl4 toxicity, thereby
increasing the inability of cirrhotic livers to metabolize
insulin.29 Thus, at low insulin infusion rates, insulin
sensitivity was normal in both acute and delayed cir-
rhotic rats, a result in agreement with Meyer-Alber et
al.,18 who did not find any difference in GIR in CCl4-
induced cirrhotic rats tested 3 to 4 days after the last
CCl4 administration and control rats. At a supraphysio-
logical insulin infusion rate (30 mU/kgrmin), there was
a striking 30% decrease of total body glucose disposal
in the acute group of cirrhotic rats, whereas the GIR
values of the delayed group were similar to controls.
These results are also in accordance with those re-
ported by Meyer-Alber et al.,18 but in sharp contrast
with those obtained in human cirrhosis, in which insu-
lin-mediated glucose disposal is mainly reduced at
physiological plasma insulin concentrations,6 and insu-
lin resistance is decreased in the supraphysiological
range of insulin perfusions.8 Our findings suggest that
the decreased insulin responsiveness30 observed in the
acute group may be more related to the CCl4-induced
acute tissular lesions than to the cirrhosis itself.
In humans, skeletal muscles are known to be respon-
sible for the disposal of most of a glucose load.31 In
rodents, however, other insulin-sensitive tissues, such
as white and brown adipose tissues, may play a more
important role.32 The hyperinsulinemic clamp does not
give any information with regard to the individual re-
sponse of insulin-sensitive tissues, which can be ex-
plored by [3H]2-deoxy-glucose injections combined with
the clamp studies.33,34 In the acute group, we found a
statistically significant decrease of glucose uptake in
all tissues tested except WAT. In the delayed group,
despite the absence of insulin resistance shown by the
clamp, glucose uptake was significantly decreased in
IBAT, diaphragm, tibialis, and gastrocnemius but not
in the other muscles of the hindlimb or in WAT. Glob-
ally, there was a tendency toward reduced glucose up-
take in muscles in the delayed group compared with
controls; however, this reduction of muscular glucose
metabolic index was more pronounced in the acute
group. This difference of insulin-mediated peripheral
tissue glucose uptake in the acute and delayed group
of cirrhotic rats may be related to the acute toxic effects
of CCl4 rather than to the effects of cirrhosis per se.
The discrepancy between the results of the clamp
studies and muscular glucose metabolic index in the
delayed group of cirrhotic rats may be because of the
lack of sensitivity of the clamp technique; differences
of less than 15% are in the range of the intra-individual
variability of the clamp.35 Another possible explanation
is that plasma insulin levels were significantly higher
in the delayed group versus controls, thus decreasing
the likelihood of identifying a slight difference in GIR.
Indeed, the difference of the ratio GIR–plasma insulin
concentration between controls and delayed cirrhosis
5p0b$$0028 02-20-96 20:13:36 hepa
MION ET AL. 587
almost reached statistical significance (0.053 { 0.002
vs. 0.046 { 0.002; P Å .072). Thus, insulin resistance
cannot be completely ruled out in the delayed cirrhosis
group.
In conclusion, our results show that impairment in
glucose metabolism in CCl4-cirrhotic rats depends on
the delay observed between the last administration of
the toxin and the metabolic studies. Acute lesions in-
duced by CCl4 administration can modify insulin sensi-
tivity; the relevance of this experimental model for the
study of glucose metabolism in liver cirrhosis is thus
questionable. Its inadequacy for the study of other met-
abolic pathways has already been suggested.36,37
Acknowledgment: The authors thank F. Berger,
M.D., for the analysis of pathological liver specimens,
M. F. Brassard and F. Lascaux (Inbiomed), for their
diligent analysis of isotopic breath samples, and M.
Rousseau, Ph.D., (Inbiomed), for her continuous sup-
port.
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