Leptin Levels in Pancreatitis 243

243
Tohoku J. Exp. Med., 2004, 204, 243-248
Received May 14, 2004; revision accepted for publication September 17, 2004.
Address for reprints: Nihat Yavuz, M.D., Caferaga Mahallesi, Moda Caddesi, Sakizgulu Sokak, No: 1/3 Daire:
15, Kadikoy, Istanbul, Turkey.
e-mail: nyavuz@tnn.net, nyavuz@istanbul.edu.tr
Plasma Leptin Levels in Rats with Pancreatitis
NIHAT YAVUZ, ETHEM UNAL,
1
KEMAL MEMISOGLU,
2
OSMAN KRAND,
1
ALI RIZA
KIZILER,
3
BIRSEN AYDEMIR,
3
RAMAZAN KUSASLAN, MUSTAFA DOGAN, PEMBEGUL
GUNES
4
and IZZET TITIZ
1
Department of General Surgery, Cerrahpasa Faculty of Medicine, Istanbul
University,
1
Department of General Surgery, Haydarpasa Numune Teaching
Hospital,
2
Department of General Surgery, PTT Erenkoy Teaching Hospital,
3
Department of Biophysics, Cerrahpasa Faculty of Medicine, Istanbul University,
4
Department of Pathology, Haydarpasa Numune Teaching Hospital, Istanbul,
Turkey
YAVUZ, N., UNAL, E., MEMISOGLU, K., KRAND, O., KIZILER, A.R., AYDEMIR, B.,
KUSASLAN, R., DOGAN, M., GUNES, P. and TITIZ, I.

Plasma Leptin Levels in Rats with
Pancreatitis. Tohoku J. Exp. Med., 2004, 204 (4), 243-248 Diagnosis of pancre-
atitis is based on the determination of serum amylase and lipase levels. However, re-
cent identifcation of specifc leptin receptors in the pancreas suggests that this pep-
tide may also play some roles in the modulation of pancreatic function. The
objective of the present study was to investigate the relationship between serum
leptin levels and pancreatitis. Thirty male Wistar rats were divided into 3 groups: the
control group, acute pancreatitis group and chronic pancreatitis group. Pancreatitis
was induced by injection of ethyl alcohol into the common biliary duct. A sham lap-
arotomy was performed in the control group. Control and acute pancreatitis groups
were sacrifced 24 hours later, and chronic pancreatitis group was sacrifced on post-
operative day 7. Blood was taken by cardiac puncture for the determination of plas-
ma leptin levels, and the pancreatic tissue was excised for histopathologic confrma-
tion of pancreatitis. Plasma leptin rose signifcantly from the median of 0.78 0.12
ng/ml in the control group to 1.92 0.10 ng/ml and 1.86 0.13 ng/ml in acute and
chronic pancreatitis groups, respectively (p < 0.001, for both). There was no signif-
cant difference in the plasma leptin levels between the acute pancreatitis group and
the chronic pancreatitis group (p > 0.05). These fndings confrm that leptin has a
role in pancreas infammation, and the infamed tissue can be the source of local pro-
duction of leptin. leptin; acute pancreatitis; chronic pancreatitis
2004 Tohoku University Medical Press
N. Yavuz et al. 244 Leptin Levels in Pancreatitis 245
MATERIALS AND METHODS
Thirty male Wistar Albino rats weighing
200-300 g were divided into 3 groups: the control
group (n = 10), acute pancreatitis group (n = 10)
and chronic pancreatitis group (n = 10). The ani-
mals were fed on standard laboratory diet and wa-
ter ad libitum before and after surgery. The study
was approved by Cerrahpasa Medical Faculty
Laboratory Animals Ethics Committee, and all
procedures with animals were performed in ac-
cordance with the guide of the Committee on
Care and Use of Laboratory Animals (CCULA
1985).
All animals were anesthetized with ether to
undergo a midline laparotomy. Acute pancreatitis
was induced by injection of 48% ethyl alcohol, in
a volume of 1 cm
3
, into the common biliary duct
using an insulin injector. A sham laparotomy was
performed in the control group.
Control and acute pancreatitis groups were
sacrifced 24 hours later, and chronic pancreatitis
group was sacrifced on postoperative day 7.
Under ether anesthesia, 4 cm
3
(3-7 cm
3
) of blood
was taken by cardiac puncture. Then, a laparoto-
my was done and pancreatic tissue was excised
and fxed in 10% formol solution for histopatho-
logic confrmation of pancreatitis. Histopa-
thological evaluation was done under light mi-
croscopy, after sectioning and staining with
hematoxylin and eosin (H & E).
Plasma leptin was measured using diagnostic
system laboratories (DSL) kit by radioimmunoas-
say (RIA) method. The concentrations were ex-
pressed as ng/ml.
All values are expressed as the mean S.E.M.
Statistics were done by SPSS program at 11.5 ver-
sion. One-way ANOVA and Scheffes F post-hoc
tests were used for multiple comparison. P < 0.05
was considered as statistically signifcant.
RESULTS
Histopathological studies confrmed alcohol-
induced pancreatitis. Infammatory infltration of
neutrophils and mononuclear cells, interstitial
Pancreatitis is an infammatory disorder of
the pancreas associated with edema, various
amounts of autodigestion, necrosis, and, in some
cases, hemorrhage. Clinically, it is defned by a
typical symptom complex associated with elevat-
ed serum amylase and lipase levels. However, in
recent years, specifc biochemical markers like
catalase, protease inhibitors, and complements
have been used for diagnostic criteria.
Leptin is known mainly for its role in the
regulation of food intake, body composition and
energy expenditure through a central feedback
mechanism (Konturek et al. 2001). In several re-
cent studies, it has also been shown that leptin has
a profound inhibitory infuence upon insulin se-
cretion by acting directly on the pancreatic beta-
cells (Jaworek et al. 2003; Perez et al. 2004). In
most overweight individuals, however, physiolog-
ical regulation of body weight by leptin seems to
be disturbed, representing leptin resistance.
This leptin resistance at the level of the pancreatic
beta-cell is suggested to contribute to the develop-
ment of hyperinsulinemia and manifest type 2 di-
abetes in overweight patients (Jaworek et al.
2003).
There are also some data in the recent litera-
ture showing that leptin plays a role in the infam-
matory process. It has been shown that leptin re-
ceptor expression on T lymphocytes modulates
chronic intestinal infammation in mice (Jaworek
et al. 2002). Leptin has also been used as an in-
fammatory marker protein to predict the cardio-
vascular disease in patients with end-stage renal
disease (Siegmund et al. 2004). Recent detection
of specifc leptin receptors in the pancreas sug-
gests that this peptide may also play some roles in
the modulation of pancreatic function (Zoccali et
al. 2004). Leptin may modulate infammatory re-
sponses in pancreatitis. However, the involve-
ment of leptin in pancreatitis is unknown.
In the present study, we aimed to detect any
possible change in the plasma leptin levels in rats
with alcohol-induced acute and chronic pancreati-
tis.
N. Yavuz et al. 244 Leptin Levels in Pancreatitis 245
edema and focal necrotic areas were seen in the
pancreatic tissues of acute pancreatitis group (Fig.
1), and interstitial fbrosis, lymphocyte infltra-
tion, ductal and ductular dilation, acinar cell atro-
phy, periductal ductular hyperplasia were seen in
chronic pancreatitis group (Fig. 2).
Plasma leptin rose signifcantly from the me-
dian of 0.78 0.12 ng/ml in the control group to
1.92 0.10 ng/ml and 1.86 0.13 ng/ml in acute
and chronic pancreatitis groups, respectively (p <
0.001, for both) (Fig. 3). However, there was no
signifcant difference in the plasma leptin levels
Fig. 2. Interstitial fbrosis, lymphocyte infltration, ductal dilatation and aciner cell atrophy showing
chronic pancreatitis. Bar, 25 m.
Fig. 1. Interstitial edema, neutrophil and mononuclear cell infltration showing acute pancreatitis.
Bar, 5 m.
N. Yavuz et al. 246 Leptin Levels in Pancreatitis 247
between the acute pancreatitis group and the
chronic pancreatitis group (p > 0.05).
DISCUSSION
Pancreatitis is an infammatory process in
the pancreas, and its diagnosis is made on the ba-
sis of the clinical presentation combined with ap-
propriate laboratory determinations, such as se-
rum amylase and lipase levels, and radiologic
fndings. However, regarding an experimental
study, histopathological confrmation is the most
important step for precise diagnosis of both acute
and chronic pancreatitis. Therefore, in the present
study, histopathological evaluation of pancreatic
tissues made it unnecessary to determine the se-
rum amylase and lipase levels.
The hormone leptin is secreted from white
adipocytes, and plasma levels of leptin correlate
with adipose tissue mass (Seufert 2004). Leptin
was frst described to act on the satiety center in
the hypothalamus through specifc receptors to re-
strict food intake and enhance energy expenditure
(Konturek et al. 2001). It has also been demon-
strated that leptin has some peripheral effects such
as inhibition of insulin biosynthesis and secretion
in pancreatic beta-cells (Jaworek et al. 2003;
Benomar et al. 2004; Perez et al. 2004). In most
overweight individuals, however, physiological
regulation of body weight by leptin seems to be
disturbed, representing leptin resistance. This
leptin resistance at the level of the pancreatic be-
ta-cell may contribute to dysregulation of the adi-
po-insular axis and promote the development of
hyperinsulinemia and manifest type 2 diabetes in
overweight patients (Jaworek et al. 2003).
Leptin has also been shown to elicit a num-
ber of immunoregulatory effects, including the
promotion of T cell proliferative responses, and
the induction of proinfammatory cytokines
(Siegmund et al. 2004; Zarkesh-Esfahani et al.
2004). Leptin defciency was shown to be associ-
ated with an increased susceptibility to infection
(Goren et al. 2003; Zarkesh-Esfahani et al. 2004).
As polymorphonuclear neutrophils (PMN) play a
major role in innate immunity and host defense
against infection, the infuence of leptin on PMN
activation was studied, and the presence of leptin
recept or i n human PMN was det ermi ned
(Caldefe-Chezet et al. 2001). These fndings pro-
vide an additional link among the obesity-derived
hormone leptin, innate immune function, and in-
fectious disease.
Recent identifcation of specifc leptin recep-
tors in the pancreas suggests that this peptide may
Fig. 3. Schematic representation of plasma leptin levels.
*
p < 0.001.
N. Yavuz et al. 246 Leptin Levels in Pancreatitis 247
also play some role in this gland (Zoccali et al.
2004). However, the involvement of leptin in
pancreatitis remains unknown. Therefore, the ob-
jective of this study was to investigate the rela-
tionship between serum leptin levels and pancre-
atitis.
In a recent study, Konturek et al. (2002) have
shown that acute pancreatitis in rats and in hu-
mans is associated with a marked increase in the
plasma level of leptin. The authors claimed that
infamed pancreas could be the source of local
production of leptin. They also showed that ex-
ogenous leptin administration protects the pancre-
as against development of acute pancreatitis. In
another study, Warzecha et al. (2002) examined
the infuence of leptin administration on the de-
velopment and the course of acute ischemic pan-
creatitis. They concluded that leptin reduces the
pancreatic damage in the course of ischemic pan-
creatitis and accelerates the pancreatic tissue re-
pair. According to the authors, the benefcial ef-
fects of leptin appear to be dependent on the
improvement of pancreatic blood fow, the in-
crease in pancreatic cell growth, and the limita-
tion of pro-infammatory interleukin-1beta release.
Jaworek et al. (2003) have also examined the
effect of exogenous leptin on pancreatic enzyme
secretion in vitro using isolated pancreatic acini,
and they concluded that leptin could take a part in
the inhibition of postprandial pancreatic secretion
and this effect could be related to the direct action
of this peptide on pancreatic acini.
For the frst time in the literature, we as-
sessed plasma leptin concentrations in both acute
and chronic pancreatitis, and our fndings showed
a signifcant rise in plasma leptin level in both
conditions (p < 0.001). However, there was no
signifcant difference between acute and chronic
pancreatitis groups (p > 0.05). These fndings
confrm that leptin has a role in pancreas infam-
mation, and the infamed tissue can be the source
of local production of leptin, because while the
infammation process continues, leptin level
seems to remain elevated.
In conclusion, leptin has both central and pe-
ripheral effects, such as restriction of food intake,
enhancement of energy expenditure, inhibition of
insulin biosynthesis, and secretion in pancreatic
beta-cells. Its immunoregulatory role has also
been documented. The present study indicates
that leptin is involved in the infammatory pro-
cesses of pancreas.
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