European Journal of Clinical Investigation (2006) 36, 528 535

Aminoguanidine prevents arterial stiffening in a new rat

model of type 2 diabetes
Blackwell Publishing Ltd

K.-C. Chang*, C.-D. Tseng, M.-S. Wu, J.-T. Liang, M.-S. Tsai, Y.-L. Cho* and Y.-Z. Tseng
*

National Taiwan University, National Taiwan University Hospital, Tapei, Taiwan, Show Chwan Memorial Hospital,

Chang-Hua, Taiwan

Abstract

Background Formation of advanced glycation end-products (AGEs) on collagen within the

arterial wall may be responsible for the development of diabetic vascular injury. This study
focused on investigating the role of aminoguanidine (AG), an inhibitor of AGE formation,
in the prevention of noninsulin-dependent diabetes mellitus (NIDDM)-derived arterial
stiffening and cardiac hypertrophy in rats.
Materials and methods The NIDDM was induced in male Wistar rats, which were
administered intraperitoneally with 180 mg kg1 nicotinamide (NA) 30 min before an
intravenous injection of 50 mg kg1 streptozotocin (STZ). After induction of diabetes
mellitus type 2, animals receiving daily peritoneal injections with 50 mg kg1 AG for 8 weeks
were compared with the age-matched, untreated, diabetic controls.
Results After exposure to AG, the STZ-NA diabetic rats had improved aortic distensibility, as
evidenced by 188% reduction of aortic characteristic impedance (P < 005). Treatment of the
experimental syndrome with AG also resulted in a significant increase in wave transit time
(+237%, P < 005) and a decrease in wave reflection factor (266%, P < 005), suggesting that
AG may prevent the NIDDM-induced augmentation in systolic load of the left ventricle. Also,
the glycation-derived modification on aortic collagen was found to be retarded by AG. The
diminished ratio of left ventricular weight to body weight suggested that prevention of the diabetesrelated cardiac hypertrophy by AG may correspond to the drug-induced decline in aortic stiffening.
Conclusions Long-term administration of AG to the STZ-NA diabetic rats imparts
significant protection against the NIDDM-derived impairment in vascular dynamics, at least
partly through inhibition of the AGE accumulation on collagen in the arterial wall.
Keywords Advanced glycation end-products, aminoguanidine, aortic input impedance,
pulse wave reflection, streptozotocin-nicotinamide diabetic rats.
Eur J Clin Invest 2006; 36 (8): 528535

Introduction
Diabetes mellitus (DM) is one of the most common
problems challenging physicians this century; approximately 140 million people world-wide currently have
diabetes, with the number projected to reach 300 million
by 2025 [1]. Type 2 DM, or noninsulin-dependent diabetes

National Taiwan University (K.-C. Chang, Y.-L. Cho), National

Taiwan University Hospital (C.-D. Tseng, M.-S. Wu, J.-T. Liang,
M.-S. Tsai, Y.-Z. Tseng), Tapei, Taiwan, Show Chwan Memorial
Hospital (Y.-Z. Tseng), Chang-Hua, Taiwan.
Correspondence to: K.-C. Chang, Department of Physiology,
College of Medicine, National Taiwan University, No. 1, Sec. 1,
Jen-Ai Road, Taipei, Taiwan. Tel.: +886 2 2312 3456; fax:
+886 2 2396 4350; e-mail: kcchang@ha.mc.ntu.edu.tw
Received 14 April 2006; accepted 19 May 2006
2006 The Authors. Journal Compilation 2006 Blackwell Publishing Ltd

mellitus (NIDDM), accounts for 90% of all cases. The

NIDDM is a complex metabolic disorder characterized by
hyperglycaemia and is associated with a relative deficiency
of insulin secretion, along with a reduced response of target
tissues to insulin. An important consequence of prolonged
hyperglycaemia is the enhanced formation of irreversible
advanced glycation end-products (AGEs) on long-lived
protein such as collagen within the arterial wall [2]. The
AGEs are a complex and heterogeneous group of compounds
that have been shown to accumulate slowly in vascular and
renal tissues with age and at a more rapid rate in diabetes
[24]. They can react with free-amino groups on an adjacent
collagen to form cross-linkages, which are thought to be one
of the central underlying processes by which they cause
damage [5]. Thus, the diabetes-related increase in AGE
accumulation on collagen in the arterial wall may contribute
to the development of certain physical changes of the blood
vessel wall [3,6,7].

Aminoguanidine and type 2 diabetic arteries

Therapeutic interventions for reducing AGE formation

should target AGE formation by reducing cross-linking
formation [8,9]. Aminoguanidine (AG), a nucleophilic
hydrazine compound, is a prototype of scavenging agents
that inhibit AGE formation and protein-protein crosslinking [10]. It has been shown to decrease methylglyoxalmediated formation of AGEs implicated as a downstream
consequence of oxidative stress, induced by mitochondrial
dysfunction in hyperglycaemia [11,12]. Aminoguanidine
therapy of rats with insulin deficiency prevents the severe
hyperglycaemia-related disorders in systolic loading condition
for the left ventricle coupled to the vasculature [13]. It has
also been reported to improve the age-related deterioration
in the pulsatile nature of blood flows in rat arteries, possibly
through inhibition of the AGE formation [14]. However,
little attention has been given to the haemodynamic response
to AG in imparting significant protection against aortic stiffening and cardiac hypertrophy in rats with NIDDM.
In 1998, Masiello et al. [15] described a new rat model
of type 2 diabetes that shared a number of features with
human NIDDM. This diabetic syndrome was experimentally
induced in adult rats administered streptozotocin (STZ)
and partially protected with a suitable dose of nicotinamide
(NA). It was characterized by moderate and stable
hyperglycaemia, glucose intolerance, altered but significant
glucose-stimulated insulin secretion, in vivo and in vitro
responsiveness to tolbutamide. An earlier report from this
laboratory demonstrated that the AGE-modulated collagen
cross-link within the arterial wall is an import factor responsible for the development of vascular injury in rats at 8 weeks
not 4 weeks after STZ and NA administration [16]. Herein,
the authors have focused on investigating the role of AG in
the prevention of NIDDM-related arterial stiffening and
cardiac hypertrophy in the diabetic syndrome, using aortic
impedance analysis [17,18]. Glycation-derived modification
on aortic collagen was also detected by sodium dodecyl
sulphate polyacrylamide gel electrophoresis (SDS-PAGE).

529

concentrations in plasma were measured by the ELISA

method (Mercodia AB, Uppsala, Sweden). The development of hyperglycaemia was confirmed by blood glucose
determination using a Surestep Test Strip (Lifescan Inc.,
Milpitas, CA). All animals were allowed free access to the
Purina chow and water and housed two or three per cage
in a 12-h light/dark-cycle animal room. The animal experiments were conducted according to the establishments
Guide for the Care and Use of Laboratory Animals, and
were approved by the Animal Care and Use Committee of
the National Taiwan University.
General surgical procedures and measurement of the
haemodynamic variables in anaesthetized rats have been
described in our previous work [19]. In brief, the rats were
anaesthetized with sodium pentobarbital (50 mg kg1, i.p.),
placed on a heating pad, intubated, and ventilated with a
Model 131 rodent respirator (New England Medical Instruments, Medway, MA). The chest was opened through the
second intercostal space of the right side. An electromagnetic
flow probe, Model 100 series of internal circumference 8 mm,
(Carolina Medical Electronics, King, NC) was positioned
around the ascending aorta to measure the pulsatile aortic
flow. A high-fidelity pressure catheter (Model SPC 320, size
2F; Millar Instruments, Houston, TX) was used to measure
the pulsatile aortic pressure via the isolated carotid artery
of the right side. The electrocardiogram (ECG) of lead II
was recorded with an ECG/Biotach amplifier (Gould,
Cleveland, OH). The selective pressure and flow signals of
510 beats were averaged in the time domain, using the
peak R-wave of ECG as a fiducial point. Timing between
the pressure and flow signals, owing to spatial distance
between the flow probe and proximal aortic pressure
transducer, was corrected by a time-domain approach, in
which the foot of the pressure waveform was realigned with
that of the flow [20]. The resulting pressure and flow signals
were subjected to further aortic impedance analysis.

Aortic input impedance spectra

Materials and methods

Animals and catheterization
Two-month-old Male Wistar rats were randomly divided
into four groups (n = 12 in each group) as follows: (i) normal
controls (NC); (ii) rats of DM type 2 (STZ-NA); (iii) NC
treated with AG (NC + AG); and (iv) STZ-NA treated with
AG (STZ-NA + AG). The NIDDM was induced in
animals, which were administered intraperitoneally
180 mg kg1 NA (Sigma, St. Louis, MO) 30 min before an
intravenous injection of 50 mg kg1 STZ (Sigma, St. Louis,
MO) dissolved in 01 M citrate buffer (pH 45) [15]. After
induction of DM type 2, the STZ-NA rats were randomized
into a vehicle-treated diabetic group, and a treatment group
receiving daily injections of 50 mg kg1 AG (Sigma, St.
Louis, MO). The animals were studied 8 weeks after being
induced with diabetes to detect the effects of NIDDM and
AG on the physical properties of the vasculature. Insulin

The aortic input impedance can be obtained from the ratio

of ascending aortic pressure harmonics to the corresponding flow harmonics, using a standard Fourier series
expansion technique (Fig. 1) [17,18]. Total peripheral
resistance of the systemic circulation (Rp) was calculated as
the mean aortic pressure/mean aortic flow. The aortic
characteristic impedance (Zc) was computed by averaging
high-frequency moduli of the aortic input impedance data
points (4th10th harmonics) [21,22]. Taking Zc into
consideration, the systemic arterial compliance (C ) was
calculated at mean aortic pressure (Pm) by expanding the
two-element [23] into the three-element Windkessel model,
which accounts for a nonlinear exponential pressurevolume relationship:
bP

SV b
e m
,
bP
bP
K + Z c SV / Ad
e ie d
where SV is the stroke volume; K is the ratio of total area
under the aortic pressure curve to the diastolic area (Ad);
C( Pm ) =

2006 The Authors. Journal Compilation 2006 Blackwell Publishing Ltd, European Journal of Clinical Investigation, 36, 528535

530

K.-C. Chang et al.

Figure 1 Aortic input impedance spectra derived from ascending

aortic pressure and flow signals from a STZ-NA diabetic rat treated
with AG (dashed line) compared with those of an untreated diabetic
animal (solid line). STZ-NA, diabetic rats at 8 weeks after being
administered STZ and NA; AG, aminoguanidine; Rp, total
peripheral resistance; Zc, aortic characteristic impedance.

b is the coefficient in the pressure-volume relation (00131

0009 of aortic arch); Pi is the pressure at the time of
incisura; and Pd is the end-diastolic pressure.
The wave transit time () can be computed by the impulse
response of the filtered aortic input impedance (Fig. 2).
This was accomplished by the inverse transformation of Zi
after multiplication of the first 12 harmonics by a DolphChebychev weighting function of the order of 24 [24].
Meanwhile, the time domain reflection factor (Rf) was
derived as the amplitude ratio of backward-to-forward peak
pressure wave method proposed by Westerhof et al. [25].
Therefore, both the wave transit time and the wave reflection
factor could characterize the wave reflection phenomenon
in the vasculature.

Gel electrophoresis
A method for measuring collagen glycation was proposed
by Turk et al. [26]. Collagen samples from aortic walls,
previously digested by pepsin, proteinase K and collagenase,
were investigated by sodium dodecyl sulphate polyacrylamide

Figure 2 Impulse response function curve derived from filtered

aortic input impedance spectra shown in Fig 1. Long arrow shows
the discrete reflection peak from the body circulation and short
arrow shows the initial peak as a reference. Half the time difference
between the appearance of the reflected peak (long arrow) and the
initial peak (short arrow) approximates the wave transit time in the
lower body circulation. STZ-NA, diabetic rats at 8 weeks after
being administered STZ and NA; AG, aminoguanidine.

gel electrophoresis (SDS-PAGE) on a Mini PROTEAN 3

System (Bio-Rad Laboratory, Hercules, CA). This was
carried out using a 4% stacking and a 10% separating gel,
running buffer system (Tris-HCl, Ph 83/SDS/glycine), and
Coomassie blue staining (Fig. 3). Each lane was loaded with
20 g protein from two or three rats. Protein blotting
analysis was carried out on PVDF membrane, using
anti-AGE antibody 6D12 (Trans Genic Inc., Kumamoto,
Japan).

Statistics
Results are expressed as means SD. A two-way analysis
of variance (anova) was used to determine the effects of
NIDDM and AG on the physical properties of the rat arterial system, using the statistical package SAS (SAS Institute
Inc., Cary, NC). Simple effect analysis was used when significant interaction between NIDDM and AG occurred.
Differences between means within levels of a factor were
determined by Tukeys honestly significant difference
(HSD) method. Significant differences were set at the level
of P < 005.

2006 The Authors. Journal Compilation 2006 Blackwell Publishing Ltd, European Journal of Clinical Investigation, 36, 528535

Aminoguanidine and type 2 diabetic arteries

531

Figure 3 Left: data showing SDS-PAGE

electrophoretic profiles of aortic collagen
from the STZ-NA diabetic rats treated with
AG (n = 2) compared with those of the
untreated diabetic animals (n = 2). Right:
corresponding protein blotting analysis on
PVDF membrane, using anti-AGE antibody
6D12. Diabetic collagen samples display
molecular weight fragments 2540 kDa, and
the glycated aortic collagen was diminished
by treatment of the experimental syndrome
with AG for 8 weeks. STZ-NA, diabetic rats
at 8 weeks after being administered STZ and
NA; AG, aminoguanidine.

Results
Table 1 shows the effects of NIDDM and AG on bloodglucose level, plasma insulin, body-weight (BW), left
ventricular weight (LVW), and aortic pressure profile in
the STZ-NA rats. The experimental syndrome yielded
a moderate and stable hyperglycaemia and prevented
STZ-induced hypoinsulinaemia and body-weight loss.
Body-weight, blood-glucose level, and plasma insulin of the
STZ-NA diabetic animals did not change in response to AG
treatment. In contrast, the significant increases in LVW and
LVW/BW ratio by prolonged hyperglycaemia were not
observed in the corresponding AG-treated group. Neither
NIDDM nor AG produced a significant change in aortic
pressure profile, nor was there an NIDDM AG interaction
for the arterial blood pressure. No significant differences in

the haemodynamic variables were observed between the

normal controls with or without AG treatment.
Figure 1 shows the effects of AG on the aortic input
impedance spectra from a STZ-NA diabetic rat. Treatment
of this experimental NIDDM with AG showed a decrease
in moduli at lower harmonics, a fall in averaging highfrequency moduli of the aortic characteristic impedance and
a decline in amplitude of the fluctuation between maximum
and minimum impedance moduli. Figure 2 describes the
impulse response function curve derived from the filtered
aortic input impedance spectra shown in Fig. 1. After
exposure to AG, the diabetic animal exhibited an increase
in wave transit time, suggesting that AG may prevent the
NIDDM-derived abnormality in the timing of the pulse
wave reflection along the path. Figure 3 demonstrates the
SDS-PAGE electrophoretic profiles of aortic collagen from

Table 1 Effects of noninsulin-dependent diabetes mellitus and aminoguanidine on body-weight, left ventricular weight, plasma glucose
and insulin level and aortic pressure profile in male Wistar rats (n = 12 per group)
Variable

NC

NC + AG

STZ-NA

STZ-NA + AG

BW (g)
LVW (mg)
LVW/BW (mg g1)
Glucose (mg dL1)
Insulin (IU mL1)
Ps (mmHg)
Pd (mmHg)
Pm (mmHg)
PP (mmHg)

4433 285
8016 507
181 009
1023 179
718 302
121 105
979 82
1067 72
233 29

4425 313
8017 595
181 007
1011 208
713 287
1173 84
934 94
1031 68
24 33

4208 303
863 605*
206 013*
1596 238*
692 280
1189 171
927 169
1077 155
259 34

4117 349
760 643
186 012
1541 256
643 275
1162 95
90 74
1048 88
255 37

All values are expressed as means SD.

*
Statistical difference (P < 005) from the control group (NC).

Statistical difference (P < 005) from the STZ-NA group.

AG, aminoguanidine; BW, body-weight; LVW, left ventricular weight; NA, nicotinamide; NIDDM, noninsulin-dependent diabetes
mellitus; Ps, systolic aortic pressure; Pd, diastolic aortic pressure; Pm, mean aortic pressure; PP, pulse pressure; STZ, streptozotocin;
STZ-NA, diabetic rats at 8 weeks after being administered STZ and NA.
2006 The Authors. Journal Compilation 2006 Blackwell Publishing Ltd, European Journal of Clinical Investigation, 36, 528535

532

K.-C. Chang et al.

the animals studied. There was a 62% increase in collagen

AGE content with prolonged hyperglycaemia and the
glycated aortic collagen was diminished by administration
of AG for 8 weeks to the STZ-NA diabetic rats.
Figure 4 shows the effects of NIDDM and AG on the
static haemodynamic data in terms of basal heart rate (HR),
cardiac output (CO), stroke volume (SV ) and total peripheral
resistance (Rp). The HR showed a significant decrease in the
rats with type 2 diabetes; however, HR did not change in
response to AG treatment (Fig. 4a). Moderate hyperglycaemia
after 8 weeks from induction markedly lowered CO
(Fig. 4b) and SV (Fig. 4c) and the NIDDM-derived alterations were retarded by AG treatment. A decrease in CO in
the absence of any significant changes in Pm (Table 1)
caused an increase in Rp in the STZ-NA diabetic animals,
from (554 68 to 660 85) mmHg s mL1 (P < 005)
(Fig. 4d). Administration of AG to this experimental
syndrome for 8 weeks prevented the diabetes-related
deterioration in physical properties of the resistance vessels,
as evidenced by the reduction of 167% in Rp (P < 005).
In contrast, the static components of the ventricular
after-load, including HR, CO, SV and Rp, were unaffected
by treatment of the normal controls with AG.
Figure 5 shows the effects of NIDDM and AG on the
pulsatile nature of blood flows in arteries using aortic
characteristic impedance (Zc), aortic compliance (Cm), wave

transit time (), and wave reflection factor (Rf). Diabetes

contributed to a marked increase in Zc, from 150 026 to
197 030 mmHg s mL1 (P < 005) (Fig. 5a) and a
decrease in Cm, from 104 15 to 82 17 L mmHg1
(P < 005) (Fig. 5b). The AG administered to the diabetic
animals for 8 weeks prevented the NIDDM-induced
decline in aortic distensibility, as manifested by the fall of
188% in Zc (P < 005) and the rise of 263% in Cm
(P < 005). Meanwhile, the experimental syndrome had
increased Rf (046 008 vs. 062 014; P < 005) (Fig. 5c)
and decreased (252 29 vs. 204 27 ms; P < 005)
(Fig. 5d). Early return with the augmented magnitude of
the reflected wave from the peripheral circulation was
retarded by treatment of the diabetic animals with AG, as
evidenced by the increase of 237% in (P < 005) and by
the reduction of 266% in Rf (P < 005). In contrast, the
oscillatory components of the ventricular after-load, including Zc, Cm, and Rf , were not modified by administration
of AG to the normal controls.

Discussion
The major findings of this study were that long-term
treatment of the STZ-NA diabetic rats imparts significant

Figure 4 Effects of NIDDM and AG on: (a) basal heart rate (HR), (b) cardiac output (CO), (c) stroke volume (SV ) and (d) total peripheral
resistance (Rp), (n = 12 per group). Treatment of the experimental syndrome with AG for 8 weeks prevented the NIDDM-related
deterioration in the physical properties of the resistance vessels, as evidenced by the reduction of 167% in Rp. By contrast, AG exerted
no effects on those static haemodynamic variables in age-matched normal controls (NC). STZ-NA, diabetic rats at 8 weeks after being
administered STZ and NA; AG, aminoguanidine.
2006 The Authors. Journal Compilation 2006 Blackwell Publishing Ltd, European Journal of Clinical Investigation, 36, 528535

Aminoguanidine and type 2 diabetic arteries

533

Figure 5 Effects of NIDDM and AG on: (a) aortic characteristic impedance (Zc), (b) systemic arterial compliance at mean aortic pressure
(Cm), (c) wave reflection factor (Rf) and (d) wave transit time (), (n = 12 per group). The NIDDM-derived alterations in the mechanical
properties of the Windkessel vessels were retarded by administration of AG to the diabetic animals for 8 weeks, as reflected in the fall of
188% in Zc and the rise of 263% in Cm. The AG also improved the systolic loading condition for the left ventricle coupled to the arterial
system by diminishing 266% of Rf and delaying 237% of . By contrast, AG exerted no effects on the pulsatile nature of blood flows in
arteries in age-matched normal controls (NC). STZ-NA, diabetic rats at 8 weeks after being administered STZ and NA; AG,
aminoguanidine.

protection against the prolonged and moderate hyperglycaemiaderived impairment in vascular dynamics; at least in part,
through inhibition of the AGE formation and accumulation within the blood vessel wall.
Herein, basal heart rate HR showed a significant decrease
in the rats of DM type 2 and HR did not change in response
to AG treatment (Fig. 4a). These results were in accordance with the report from Schmidt et al. [27], who found
that long-term administration of AG to diabetic animals
could not prevent the development of sympathetic neuroaxonal dystrophy, which is responsible for the decrease in
HR.
A decline in cardiac output occurred in the absence of
any significant changes in mean aortic pressure, causing a
rise in total peripheral resistance (Rp) in the STZ-NA
diabetic rats. It has been shown that prolonged hyperglycaemia may enhance the formation of both reversible
amadori intermediate products and irreversible AGEs within
the artery wall [2,4]. The AGEs are reported to induce freeradical production and deplete nitric oxide (NO) concentration, leading to a state of oxidative stress [28]. The ability
of AGEs to quench NO is supposed to diminish the
vasodilatory capacity of the peripheral muscular arteries in
the rats of type 2 diabetes. Therefore, an increase in AGE
accumulation in the STZ-NA diabetic animals may be one

of the several factors responsible for the increased vascular

smooth muscle tone. The NIDDM-derived physical
changes in the resistance vessels were prevented by administration of AG to rats for 8 weeks, as reflected in the
reduction of 167% in Rp. In addition to being an AGE-blocker,
AG could act as a preventative agent in diabetic cardiovascular complications by NO synthase-inhibition pathways
[29]. However, a beneficial effect of AG on resistance to
blood flow in the absence of any significant change in mean
aortic pressure was observed in this report when AG was
administered to the experimental syndrome. These findings
are in agreement with those obtained by Huijberts et al.
[21], who found that arterial blood pressure remained
unaltered and aortic impedance was lower in the AG-treated
diabetic animals. Thus, the prevention of NIDDM-induced
vasodilatory dysfunction may result from inhibition of the
AGE formation by AG to reserve NO production in the
resistance vessels.
As for the pulsatile nature of blood flows in arteries, the
aortic characteristic impedance increased (Zc in Fig. 5a) and
the wave transit time decreased ( in Fig. 5d) in the STZNA diabetic rats. Although being affected by the lumen
radius, Zc is inversely related to the aortic distensibility such
that Zc has been frequently used as an indicator of aortic
stiffness: the higher the aortic characteristic impedance, the

2006 The Authors. Journal Compilation 2006 Blackwell Publishing Ltd, European Journal of Clinical Investigation, 36, 528535

534

K.-C. Chang et al.

stiffer the aortic wall [17,18]. Meanwhile, being relatively

independent to body shape, wave transit time (), which is
inversely related to pulse wave velocity, could also be derived
to describe the aortic distensibility: the stiffer the aortic wall,
the shorter the wave transit time and vice versa [17]. Herein,
both the augmented Zc and the shortened suggest that a
decline in aortic distensibility may occur in the STZ-NA
diabetic animals. It has been shown that accumulation of
AGEs in the arterial wall is associated with changes in the
biomechanical properties of collagen characterized by
increasing stiffness of the elastic reservoir [4,10]. There
was a 62% increase in aortic collagen AGE content in the
rats of type 2 diabetes. So, the AGE-modulated collagen
cross-link in the Windkessel vessels could function as
one of the several factors to increase the arterial stiffness
with diabetes. The NIDDM-induced fall in aortic distensibility was prevented by 8 weeks treatment of the
experimental syndrome with AG, as manifested by the
reduction of 188% in Zc and the increase of 237% in .
The AG treatment also contributed to a rise of 263% in
systemic arterial compliance of the rats with prolonged
hyperglycaemia (Fig. 5b). Such prevention of the NIDDMderived arterial stiffness by AG was supported by the fact
that the glycated collagen in the aortic wall was diminished
by administration of AG to this new rat model of type 2 diabetes (Fig. 3).
Interestingly, no significant changes in arterial pulse
pressure were observed in the STZ-NA diabetic animals
with the increased arterial stiffness (Table 1). Several factors
importantly affect the magnitude of the pulse pressure,
including the stroke volume and the aortic compliance [30].
The arterial pulse pressure varies directly with the stroke
volume but inversely with the arterial compliance. Thus, the
decrease in SV (Fig. 4c) may blunt the effect of the diminished Cm (Fig. 5b) on the pulse pressure in the rats with
type 2 DM. After exposure to AG, this experimental
syndrome also showed no significant alteration in arterial
pulse pressure because of its counter-balancing influences
of increasing SV and augmenting Cm on this haemodynamic
parameter.
Changes in timing and/or magnitude of the pulse wave
reflection do impair the loading condition for the left
ventricle when coupled to its arterial system [31]. With the
enhanced wave reflection factor (Rf in Fig. 5c), the experimental syndrome contributed to a reduction in wave transit
time ( in Fig. 5d) in the STZ-NA diabetic rats. The
increased Rf with shortened indicates that prolonged and
moderate hyperglycaemia can modify the timing and
magnitude of the pulse wave reflection to augment systolic
load of the left ventricle. The impaired systolic loading
condition of the left ventricle may cause the heart to adapt
to muscular hypertrophy, as evidenced by the increase in
the ratio of LVW : BW (Table 1). After exposure to AG,
the diabetic animals showed a significant fall of 266% in
Rf , suggesting that the heavy reflection phenomenon may
be alleviated in the circulatory system. The observed
increase in and decrease in Rf indicate that AG, by
preventing the AGE accumulation on collagen in the
diabetic arterial wall, can improve the systolic loading

condition for the left ventricle coupled to the vasculature.

Moreover, the ratio of LVW : BW was decreased by AG
treatment, suggesting that the prevention of NIDDMrelated cardiac hypertrophy may correspond to the druginduced decline in arterial load.
This report has demonstrated that AG ameliorates
vascular complications observed in the experimentallyinduced diabetes type 2 at least partly through inhibition of
the AGE formation. However, when using AG it is important
to be aware of the facts that this drug, apart from preventing
formation of AGEs, also inhibits diamine oxidase (DAO)
[32]. In the in vivo situation inhibition of DAO might lead
to serious vascular and respiratory side-effects owing to
accumulation of histamine in the blood stream. In addition,
AG in high doses may bind to S-adenosylmethionine
decarboxylase (SAMDC) and thereby might affect polyamine
formation. The effects of AG on the mechanical properties
of the vasculature owing to inhibition of DAO and stabilization of SAMDC remains to be determined in rats treated
with STZ and NA.
The authors contribution to this endeavour was to
provide a path to consider the clinical application of an AGE
inhibitor in the prevention of NIDDM-related deterioration
in vascular dynamics. The AG may target elastic arteries
(characteristic impedance), muscular arteries (wave reflection factor) and arterioles (peripheral resistance) by inhibiting the formation of AGEs in the diabetic blood vessel wall.
However, an important limitation of AG and other AGEinhibitors is that they cannot reverse pre-existing AGE
cross-linking, and thus the AGE breakers are supposed
to be noted. Recently, several studies have used the AGE
breaker compound 4,5-dimethyl-3-phenacylthiazolium
chloride (also called ALT-711) for animal and human
studies [9,33]. The results of ongoing clinical studies will
determine if these compounds can become the first specific
therapy for the diabetes-related cardiovascular disorder.
Taken together, it has been demonstrated that moderate
and stable hyperglycaemia produces a detriment to the
physical properties of the resistance vessels and the
Windkessel vessels in rats administered STZ and NA after
8 weeks. Prevention of the NIDDM-induced vasodilatory
dysfunction may in part result from inhibition of the
AGE formation by AG to reserve NO production in the
resistance vessels. Treatment of the experimental syndrome
with AG for 8 weeks may prevent the AGE-modulated
collagen cross-link in the arterial walls so that the aortic
distensibility can be improved in the STZ-NA diabetic
animals. Moreover, AG can retard the NIDDM-derived
augmentation in systolic load of the left ventricle by
delaying the return of the pulse wave reflection associated
with the reduced wave reflection intensity. The prevention
of diabetes-related cardiac hypertrophy by AG may
correspond to the drug-induced decline in left ventricle
after-load. We concluded that long-term treatment of
the STZ-NA diabetic rats with AG imparts significant
protection against the NIDDM-derived impairment in
vascular dynamics, at least partly through inhibition of
the AGE accumulation on collagen within the arterial
wall.

2006 The Authors. Journal Compilation 2006 Blackwell Publishing Ltd, European Journal of Clinical Investigation, 36, 528535

Aminoguanidine and type 2 diabetic arteries

Acknowledgements
This study was supported by grants from the National
Taiwan University Hospital (NTUH 94-S017) and from the
National Science Council of Taiwan (NSC 94 2320-B002058).

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2006 The Authors. Journal Compilation 2006 Blackwell Publishing Ltd, European Journal of Clinical Investigation, 36, 528535