Plasma visfatin levels in patients with newly diagnosed and
untreated type 2 diabetes mellitus and impaired
Teoman Dogrua,*, Alper Sonmeza, Ilker Tascia, Ergun Bozoglua, Mahmut Ilker
Yilmazb, Halil Genca, Gokhan Erdema, Mahmut Goka, Necati Bingolc,
Selim Kilicd, Taner Ozgurtase, Sezin Bingolf
aDepartment of Internal Medicine, Gulhane School of Medicine, Etlik, Ankara, Turkey
bDepartment of Nephrology, Gulhane School of Medicine, Etlik, Ankara, Turkey
cBayındır Hospital, Biochemistry Laboratory, Ankara, Turkey
dDepartment of Epidemiology, Gulhane School of Medicine, Etlik, Ankara, Turkey
eDepartment of Biochemistry, Gulhane School of Medicine, Etlik, Ankara, Turkey
fNumune Hospital, Biochemistry Laboratory, Ankara, Turkey
Received 2 May 2006; accepted 31 July 2006
Available online 7 September 2006
Visfatin, a new adipokine, facilitates adipogenesis and has insulin-mimetic properties. We aimed to investigate the plasma
visfatin levels in patients with newly diagnosed and untreated type 2 diabetes mellitus (T2DM) and impaired glucose tolerance
(IGT), who had no obesity or hypertension. Twenty-two patients with T2DM, 18 subjects with IGT and 40 healthy controls were
enrolled. Visfatin levels were measured along with the BMI, blood pressure, lipids, glucose, insulin, adiponectin and hsCRP levels,
and HOMA-IR indexes. Age, sex and BMI were similar in all groups. Visfatin levels were higher in the diabetic group than the
controls (p = 0.01). There was no significant difference in the visfatin levels between the T2DM and IGT groups as well as IGT
group and healthy controls. Plasma visfatin concentrations did not differ between men and women. Visfatin levels did not correlate
with BMI, blood pressure, plasma adiponectin, insulin, hsCRP, glucose and lipid levels or HOMA-IR indexes in the three groups.
this increase gets more prominent as the glucose intolerance worsens.
# 2006 Elsevier Ireland Ltd. All rights reserved.
Keywords: Visfatin; Adiponectin; Type-2 diabetes mellitus; Impaired glucose tolerance
Adipose tissue has been shown to secrete a variety
of hormones, cytokines, growth factors and other
adipokines – into the circulation that can have
profound effects on the vasculature and metabolism
. These compounds include leptin, tumour necrosis
factor-alpha (TNF-a), non-esterified fatty acids,
prostaglandins, adiponectin, resistin, angiotensino-
gen, interleukin (IL)-1 and -6, and plasminogen
Diabetes Research and Clinical Practice 76 (2007) 24–29
* Corresponding author at: GATA Ic Hastaliklari B.D., Etlik, 06018
Ankara, Turkey. Tel.: +90 312 3044036; fax: +90 312 3044000.
E-mail address: email@example.com (T. Dogru).
0168-8227/$ – see front matter # 2006 Elsevier Ireland Ltd. All rights reserved.
Visfatin is a protein that is preferentially produced in
visceral adipose tissue . It is expressed in the isolated
subcutaneous adipose cells as well. Both tissue
expression and plasma levels of visfatin increase in
parallel with obesity. It can also be found in skeletal
muscle, liver, bone marrow and lymphocytes , where
it was initially identified as pre-B-cell colony-enhan-
cing factor (PBEF). It has insulin-mimetic effects and
lowers plasma glucose levels . Moreover, circulating
visfatin concentrations were shown to increase in
parallel with hyperglycemia . However, the data
regarding pathophysiological role of visfatin in glucose
homeostasis is limited.
Elevated plasma visfatin levels have recently been
reported in a group of patients with type 2 diabetes
mellitus (T2DM) on hypoglycemic treatment .
Another study showed higher circulating visfatin levels
as well as adipose tissue visfatin mRNA expression in
diabetics compared to body mass index (BMI)
unmatched controls . Given the profound effects
of drugs and obesity on insulin sensitivity and
inflammation, plasma visfatin concentrations can
possiblybe changedinsuch patients when the treatment
is started. Therefore, in the present work, we
investigated the plasma visfatin levels in subjects with
newly diagnosed and previously untreated T2DM who
had no confounding factors for insulin sensitivity and
inflammation. We also searched the changes in plasma
visfatin concentrations in different disordered glucose
metabolism. Finally, whether there was a correlation of
visfatin with adiponectin, BMI, blood pressure, high
sensitive C-reactive protein (hsCRP), lipid levels and
insulin sensitivity was also evaluated.
2. Materials and methods
The participants of the study wereselected from the people
who were referred to the Department of Internal Medicine,
Gulhane School of Medicine, Turkey. Between June 2005 and
February 2006, 22 patients with diabetes and 18 patients with
IGT were enrolled in the study. Both of these subjects were
newly diagnosed and previously untreated.
Exclusion criteria were sustained hypertension, heart fail-
ure, peripheral vascular disease, smoking, acute or chronic
infections, cancer, hepatic or renal disease, and medication.
T2DM patients with acute illness, recent (<6 months) myo-
cardial infarction, unstable angina or stroke were also
excluded from the study. The control group consisted of 40
healthy people who had no family history of T2DM. Before
inclusion, all the study and control subjects underwent careful
physical examination and detailed laboratory investigations to
exclude any condition that may interfere with glucose toler-
ance. Standing height and body weight were measured in light
indoor clothing without shoes. Body mass index (BMI) was
calculated as weight divided by squared height (kg/m2).
The Ethical Committee of Gulhane Medical School
approved the study. All subjects gave written informed con-
sent before participating in the study.
2.2. Laboratory analyses
The blood samples were collected between 08:00 and
08:30 a.m. after a 12 h fasting. The tubes were promptly
centrifuged, and the plasma was separated and stored at
?80 8C. All samples were run in the same assay.
Plasma visfatin levels were determined by ELISA method
(Human visfatin ELISA kit, Phoenix Pharmaceuticals, Bel-
mont, CA, USA) [sensitivity: (minimum detectable concen-
tration) = 0.5–1 ng/ml, IntraCV: 5% and InterCV: 12%].
Plasma adiponectin concentrations were measured in dupli-
cate by RIA method (Human adiponectin RIA kit, Linco
research, Inc., St. Charles, MO, USA) [sensitivity: (minimum
detectable concentration) = 1 ng/ml, IntraCV: 3.59% and
Fasting plasma glucose (FPG), total cholesterol (TC),
triglyceride, and high density lipoprotein (HDL) cholesterol
levels were measured by the enzymatic colorimetric method
with Olympus AU 600 auto analyzer using reagents from
Olympus Diagnostics, GmbH (Hamburg, Germany). Low
density lipoprotein (LDL) cholesterol level was calculated
by Friedwald’s formula . The serum basal insulin level was
determined by the coated tube method (DPC, Los Angeles,
Serum hsCRP, a well-known marker of inflammation, was
determined by turbidimetric fixed rate method [A. Wasunna,
Eur J Pediatr, 1990] by an automated analyzer (Olympus AU-
2700, Mishima, Japan).
The insulin sensitivity was determined by Homeostasis
Model Assessment Model (HOMA) index with formula:
HOMA-IR = fasting insulin (mU/ml) ? fasting glucose (mg/
dl)/405 . Low HOMA-IR values indicate high insulin
sensitivity, whereas high HOMA values indicate low insulin
sensitivity (insulin resistance).
Oral glucose tolerance test (OGTT) was performed after
10–12 h of overnight fasting by ingesting 75 g of oral glucose
load over a 2-min period, and obtaining blood samples at
baseline and 2 h after glucose load for serum glucose mea-
surements. Glucose tolerance status was defined as normal
glucose tolerance (NGT), IGTor diabetes according to WHO
2.3. Statistical analysis
Results are reported as the mean ? S.D. One sample
Kolmogorov–Smirnov test was used to evaluate the distribu-
tion characteristics of variables. The differences between the
groups were tested for significance by one-way ANOVA test,
chi-square test and Kruskall–Wallis test. Tukey’s test and
T. Dogru et al./Diabetes Research and Clinical Practice 76 (2007) 24–2925
Bonferroni adjusted Mann–Whitney U-test were used as post
hoc analysis as appropriate. The relationship between vari-
ables was analyzed by Pearson’s correlation or Spearman’s
rho correlation tests as appropriate. Differences and correla-
tions were considered significant at p < 0.05.
The characteristics of the study population and the
controls are given in Table 1. Age, sex and BMI
distributions were similar among the three groups.
Plasma visfatin levels were higher in the diabetic group
than the controls (p = 0.01). No difference was
observed between men and women regarding visfatin
levels (p = 0.28). Therewas no difference in the plasma
levels of visfatin between the T2DM and IGT and also
between IGT and healthy controls (p = 0.33 and 0.47,
lower in the diabetic and IGT groups when compared to
controls (p < 0.001, for both). In addition, adiponectin
levels were lower in the diabetic group than the IGT
group (p < 0.001) (Fig. 2).
FPG, insulin and HOMA-IR indexes in the T2DM
group and only FPG in the IGT group increased
significantly compared with NGT group. hsCRP levels,
FPG, insulin levels and HOMA-IR indexes in T2DM
were significantly higher than those in IGT group, but
no significant differences in TC, HDL and triglyceride
levels were detected. HDL-C levels in the controls were
similar with the diabetic group, but higher than IGT
group. There were no differences in TC and triglyceride
levels among the three groups (Table 1).
We next analyzed the correlation between plasma
levels of visfatin and other parameters. Plasma visfatin
in the study subjects and controls did not correlate with
BMI, blood pressure, adiponectin, hsCRP, insulin,
glucose and lipid levels or HOMA-IR indexes. FPG
negatively correlatedwith adiponectinmeasurements in
the T2DM group (r = ?0.54, p = 0.01).
T. Dogru et al./Diabetes Research and Clinical Practice 76 (2007) 24–2926
The clinical characteristics and laboratory results of subjects with T2DM, IGT and healthy controls
T2DM (n = 22) IGT (n = 18)Controls (n = 40)p p1p2 p3
Systolic blood pressure
Diastolic blood pressure
Total cholesterol (mg/dl)
Fasting glucose (mg/dl)
49.6 ? 12.0
28.1 ? 4.0
125 ? 9.1
50.5 ? 10.6
28.2 ? 4.3
127.5 ? 7.7
46.9 ? 7.1
27.6 ? 3.8
127.3 ? 9.7
79 ? 9.0 81.4 ? 6.8 79 ? 6.9 0.50NS NSNS
187.2 ? 45.4
46.0 ? 8.4
215.6 ? 40.8
43.0 ? 6.8
208.6 ? 36.4
49.6 ? 9.3
Resultsareexpressedasmean ? S.D.,median(range).p1,forT2DMandIGT;p2,forT2DMandcontrols;p3,forIGTandcontrols.Tukey’stestand
Bonferroni adjusted Mann–Whitney U-test were used as post hoc test as appropriate. T2DM, type 2 diabetes mellitus; IGT, impaired glucose
tolerance; BMI, body mass index; HDL, high density lipoprotein; LDL, low density lipoprotein; HOMA-IR, homeostasis model assessment-insulin
resistance; hsCRP, high sensitive C-reactive protein.
aOne-way ANOVA test.
Fig. 1. Comparisonof visfatin levels in subjects with T2DM, IGTand
healthy controls. Visfatin levels were higher in patients with T2DM
(32.2 ? 2.3 ng/ml) compared to IGT (30.0 ? 2.9 ng/ml) and controls
(28.5 ? 2.07 ng/ml), whereas no difference was found between
T2DM vs. IGT or IGT vs. controls. Results are expressed as
mean ? S.D. T2DM: type 2 diabetes mellitus; IGT, impaired glucose
This study was designed to investigate the plasma
visfatin levels in patients with newly diagnosed and
never treated T2DM and IGT who had no accompany-
ing conditions such as obesity and hypertension, and
were free of any medication including over the counter
drugs. We found that plasma visfatin levels were higher
and adiponectin levels were lower in the diabetic
population compared to healthy controls. In addition, in
comparison with controls, circulating visfatin levels
were similar but adiponectin levels were lower in the
IGT group. No correlation was observed between
visfatin levels and the other parameters including
Adipocytes are important endocrine cells, which
secrete several hormones and cytokines that are
involved in metabolic diseases such as obesity and
T2DM. Adipose tissue derived cytokines such as TNF-
a, IL-6, hsCRP, leptin, adiponectin and resistin
contribute to the development of insulin resistance as
well as diabetes mellitus . Visfatin, a novel
adipokine having insulin sensitizing properties additive
to the effect of insulin, was reported to activate the
insulin receptors in various cell types, increase glucose
transport and lipogenesis in 3T3-L1 adipocytes or L6
myocytes, and decrease glucose production by hepa-
tocytesinvitro.Itnotonlyacutelylowers the plasma
results in decreased glucose and insulin levels when
administered to diabetic mice, an effect that is mediated
by the insulin receptor itself with similar affinity but
through binding a distinct site. Taken together, these
findings suggest that visfatin could play a role in the
studies which have examined the expression of visfatin
in diabetes mellitus; therefore, the regulation of
visfatin/PBEF in different stages of glucose intolerance
is still not entirely clear.
In the present study, the subjects with T2DM showed
significantly elevated plasma visfatin concentrations
when compared to BMI matched healthy controls.
These findings support the results of the study of Chen
et al. who reported that visfatin levels are altered in
patients with T2DM. In this study, a decrease in
adiponectin and an increase invisfatin were observed in
type 2 diabetic patients and the authors suggested that
the increasing concentrations of visfatin were indepen-
dently and significantly associated with T2DM .
However, the patients in that study were taking avariety
of hypoglycemic drugs alone or in combination, which
might possibly influence the results, though the net
effect of medication on plasma visfatin is not yet
known. In our study, we enrolled only the people with
medication. Moreover, the present study addresses the
natural behavior of visfatin levels in type 2 diabetics at
the time of diagnosis and excludes the effects of
hypoglycemic medication. In another study with a
relatively small sample size, plasma visfatin concentra-
tion as well as adipose tissue visfatin mRNA levels was
reported to be higher in patients with T2DM when
compared to healthy controls . Though the difference
between the BMIs in the two groups was a limitation of
that study, blood visfatin measurements were in line
with our results. As in the study of Chen et al. , we
found no correlation of plasma visfatin with BMI in the
present investigation. Thus, the two previous reports
and the present investigation may suggest that plasma
visfatin increases in T2DM regardless of taking
hypoglycemic medication or presence of obesity. In
addition, in order to observe the alterations in plasma
visfatin as the glycemia dysregulation progresses, we
searched whethertherewas adifference betweenpeople
with IGT and T2DM regarding plasma visfatin. We
found similar plasma visfatin concentrations in these
two groups. On the other hand, there was also no
IGT group and the controls suggesting that visfatin
might be involved in the pathogenesis of T2DM but not
at the early stages.
Adiponectin, a well-known adipocyte-derived hor-
mone, which is involved in the regulation of insulin
resistance and contributes to the glucose homeostasis,
displays protective roles in atherosclerosis and is
inversely related to the measures of cardiovascular
diseases [11–13]. It was reported to be lower in patients
with T2DM and IGTwhen compared to healthy people
, and suggested to be a predictive factor for the
T. Dogru et al./Diabetes Research and Clinical Practice 76 (2007) 24–2927
Fig. 2. Plasma adiponectin was found to decrease as the glycemia
dysregulation worsened. It was the lowest in patients with T2DM [5.1
(2.7–7.0) mg/ml],intermediateinIGT[11.4(5.8–16.3) mg/ml]andthe
as median (range). T2DM, type 2 diabetes mellitus; IGT, impaired
development of glucose intolerance . Besides
serving as a positive control measure in the present
study, since its association with several parameters
including insulin sensitivity has been well described in
certain conditions , measurement of plasma
adiponectin could be helpful in interpretation of the
results. In our investigation, plasma adiponectin
concentration was the lowest among diabetic subjects,
highest in the healthy controls, and intermediate in the
IGT group, which is in line with the previous reports
[14,17]. Plasma visfatin and adiponectin concentrations
were found not to correlate in T2DM [5,6]. In a
population with a wide range of obesity, including 36
subjects with T2DM and 31 with IGT, Berndt et al.
levelsand insulin sensitivity as well . In parallel,we
also did not find any correlation between plasma
visfatin and adiponectin levels or HOMA-IR indexes
either in diabetics or subjects with IGT. Though the
number of the study population was low in our work, it
may be suggested that the mechanisms controlling the
metabolism of these two adipocyte-derived proteins
differ in patients with hyperglycemia.
Insulin resistance and chronic sub-clinical inflam-
mation are strongly associated with T2DM and IGT
[19–21]. Visfatin is also suggested as an inflammatory
cytokine that is produced and released by the adipose
in acute lung inflammation and sepsis, which is
accompanied by an insulin-resistant state [23,24].
Interestingly, visfatin expression is regulated by
substances such as lipopolysaccharide, IL-1b, TNF-a
and IL-6 that promote insulin resistance [25,26]. In the
present study, significant, albeit weak, difference was
found in hsCRP levels between subjects with T2DM
and IGT. However, plasma visfatin was not correlated
with hsCRP levels in these two groups, suggesting that
inflammation may not take part in the mechanism of
increased visfatin concentrations in T2DM.
This study has several limitations. As a consequence
of narrow selection criteria, the numbers of the patients
werehardlyenough tomake clearestimations.Also, itis
imperative to mention that the HOMA formula used for
and may not be as accurate as glucose clamp test.
In conclusion, the results of the present study
indicate that plasmavisfatin levels are higher inpatients
with newly diagnosed and untreated T2DM when
compared to subjects with IGTor BMI matched healthy
controls. Plasma visfatin seems not to correlate with
BMI, adiponectin, hsCRP and insulin sensitivity.
Further studies with larger population are needed to
evaluate the role of visfatin in the pathogenesis of
This work was supported by grant (AR-2005/46)
from the Research Center of Gulhane Medical School.
The authors express their appreciation to Fevziye Ozar
and Burcu Gonen for their assistance.
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