Environmental Toxicology and Pharmacology 9 (2001) 71 – 78

[Link]/locate/etap

Characterization of low dose streptozotocin-induced progressive

diabetes in mice
Mikio Ito *, Yoichi Kondo, Akiko Nakatani, Koji Hayashi, Atsuhiko Naruse
Laboratory of Analytical Pharmacology, Faculty of Pharmacy, Meijo Uni6ersity, 150 Yagotoyama, Tenpaku-ku, Nagoya 468 -8503, Japan

Received 12 April 2000; received in revised form 13 October 2000; accepted 20 October 2000

Abstract

Our previous study indicated that a single i.p. injection of 100 mg/kg streptozotocin (STZ) is able to induce slowly progressive
diabetes mellitus in adult ICR mice. In the present study, the non-fasting serum insulin levels of the mice administered 100 mg/kg
STZ were normal throughout the 24-week-observation after STZ injection. In the STZ-administered mice, the area of islets and
the number of insulin-immunoreactive cells (b-cells) were normal at 1 week and then continued to decrease gradually as the time
went on. In contrast, there was a relative increase in the number of glucagon-immunoreactive cells (a-cells) in these mice. In
addition, in the STZ-administered mice, the degree of glucose tolerance continued to reduce from 2 weeks till 12 weeks when the
experiment terminated. The rise in serum insulin levels stimulated by glucose in the STZ-administered mice began to subside from
about 2 weeks and had completely ceased by 12 weeks. These results indicate that 100 mg/kg STZ-induced diabetic mouse model
is non-insulin-dependent diabetes, which is characterized by impaired insulin response to glucose stimulation. © 2001 Elsevier
Science B.V. All rights reserved.

Keywords: Streptozotocin; Slowly progressive diabetes; Non-insulin-dependent; Mouse

1. Introduction Recently, we succeeded in creating a new mouse

model of slowly progressive diabetes mellitus with only
Diabetes mellitus is classified into two types, insulin- a single i.p. injection of a subdiabetogenic small dose
dependent (IDDM, type 1) and non-insulin-dependent (100 mg/kg) of STZ in 8-week-old male ICR mice (Ito
diabetes mellitus (NIDDM, type 2). NIDDM is a dis- et al., 1999). We concluded that the diabetic model
ease characterized by peripheral insulin resistance and induced by the injection of the 100 mg/kg STZ, unlike
impaired insulin secretion. NIDDM patients are di-
200 mg/kg STZ-induced diabetes which is insulin-de-
vided into two categories, obese type with hyperinsu-
pendent, is non-insulin-dependent, because non-fasting
linemia and non-obese type with hypoinsulinemia.
Streptozotocin (STZ; N-nitroso derivative of glu- blood insulin levels were normal throughout the 9-
cosamine) is a broad-spectrum antibiotic extracted from week-observation period after administration of STZ.
Streptomyces acromogenes (Herr et al., 1959). It is a In our previous report, we did not yet present
pancreatic b-cell toxin that induces rapid and irre- changes in body weight, urine volume, and drinking
versible necrosis of b-cells (Junod et al., 1967) and is water and food consumption after administration of
widely used in making experimental animal models of STZ at various doses. In addition, it remains unclear
IDDM (Rakieten et al., 1963; Brodsky and Logotheto- whether this progressive diabetic mouse model induced
poules, 1969; Junod et al., 1969; Rerup, 1970). Junod et by a single i.p. injection of 100 mg/kg STZ is abated or
al. (1969) reported that the i.v. injection of 25–100 becomes worse after 9 weeks. Therefore, in this paper,
mg/kg STZ to rats was able to produce a dose-depen- we first report unpublished or additional data related to
dent hyperglycemia. our previous study. In order to clarify further charac-
* Corresponding author. Tel.: + 81 52 832 1781; fax: +81 52 834
teristics of this new mouse model, we followed up
8090. changes in islet morphology (the number of a- and
E-mail address: mitoh@[Link] (M. Ito). b-cells) and serum glucose and insulin responses to oral

1382-6689/01/$ - see front matter © 2001 Elsevier Science B.V. All rights reserved.
PII: S1382-6689(00)00064-8
72 M. Ito et al. / En6ironmental Toxicology and Pharmacology 9 (2001) 71–78

glucose load after STZ administration. We also exam- Tokyo, Japan) and Detamina TC-5 (Kyowa Medix,
ined the effects of tolbutamide, an agent that stimulates Tokyo, Japan), respectively.
insulin secretion, and troglitazone, an agent that im-
proves insulin resistance, on glucose tolerance in mice 2.4. Immunocytochemistry
with 100 mg/kg STZ-induced diabetes in comparison
with the effects in mice with 200 mg/kg STZ-induced For glucagon and insulin immunostaining, the mice
diabetes as well as normal mice. administered 100 and 200 mg/kg STZ and normal mice
were sacrificed by decapitation and their pancreata
were taken at 1, 4, 8, 12 and 24 weeks after STZ
2. Materials and methods administration. The pancreata were fixed in 10%
buffered formalin and then embeded in paraffin. Two
2.1. Animals consecutive sections (section thickness, 1 mm) were cut
from the paraffin block. For observation of glucagon
8-week-old male ICR mice (Nippon SLC, Shizuoka, immunoreactive cells (a-cells), one of the two sections
Japan) were housed in an isolator caging system in was stained using anti-glucagon monoclonal antibody
air-conditioned animal room at 23°C91°C. (Histofine SAB-PO kit, Nichirei, Tokyo, Japan). For
observation of insulin immunoreactive cells (b-cells),
2.2. Induction of diabetes the other section was stained by using anti-insulin
monoclonal antibody (Histofine SAB-PO kit, Nichirei,
Mice were fasted for 20 h before diabetes was in- Tokyo, Japan). The number of glucagon- and insulin-
duced with STZ. They received a single i.p. injection of immunoreactive cells in the islets was counted.
75, 100, 150 or 200 mg/kg STZ (Sigma, St. Louis, MO,
USA) freshly dissolved in 0.05 M citrate buffer, pH 4.5. 2.5. Oral glucose tolerance test
Normal mice were injected with the equivalent volume
of citrate buffer. Oral glucose tolerance test in the mice administered
100 mg/kg STZ and in normal mice was carried out on
2.3. Serum glucose, insulin and total cholesterol le6els, 3 days and 1, 2, 4, 5, 7, 9 and 12 weeks after STZ
body weight, urine 6olume, and drinking water and administration. These animals were fasted for 18 h
food consumption before the test and then given 2 g/kg glucose solution
orally. Blood samples were taken from the cavernous
In the first experiment, blood samples from normal sinus with a capillary under ether anesthesia at 0.5 h
mice and mice administered 75, 100, 150 and 200 mg/kg before and at 0.5, 1, 2 and 4 h after glucose loading.
STZ were taken from the cavernous sinus with a capil- Serum glucose and insulin levels (only at 3 days and at
lary under ether anesthesia at 1, 3, 5, 7 and 9 weeks 1, 2 and 12 weeks) were determined as described above.
after STZ administration for the determination of
serum glucose, insulin (normal mice and mice adminis- 2.6. Effects of tolbutamide and troglitazone on glucose
tered 100 and 200 mg/kg STZ) and total cholesterol. tolerance
The body weight of each mouse was measured immedi-
ately before blood collection. After collection of blood Effects of tolbutamide and troglitazone on glucose
samples, these animals were kept in individual tolerance in the mice administered 100 or 200 mg/kg
metabolic cages for 24 h and drinking water (distilled STZ and in the normal mice were evaluated at 12 weeks
water) and food consumption per 24 h were measured. after STZ administration. These animals were fasted for
In the second experiment, blood samples from mice 18 h and tolbutamide or troglitazone suspended in 1%
administered 100 and 200 mg/kg STZ were taken at 20 gum arabic solution was given orally. The control
and 0.5 h before and at 2, 7, 24, 48 and 72 h after STZ animals were given 1% gum arabic solution, instead of
administration and serum glucose and insulin were then the test drug. 1 h later, a 2 g/kg glucose was given
measured. orally. Blood samples were collected at 0.5 h before and
In the third experiment, serum glucose, insulin and 0.5, 1, 2 and 4 h after glucose loading. Serum glucose
total cholesterol, body weight, urine volume, and drink- levels were determined as described above.
ing water and food consumption of mice administered
100 and 200 mg/kg STZ and of normal mice were 2.7. Statistical analysis
measured at 12 and 24 weeks after STZ administration.
Serum glucose, insulin and total cholesterol were Results obtained were expressed as the mean9SE.
determined using commercial reagents of Glucose CII- The data were analyzed by one-way analysis and Dun-
test Wako (Wako Pure Chemical Industries, Tokyo, can multiple range test or non-parametric statistics. In
Japan), ELISA Insulin kit (Seikagaku Industries, all cases, PB 0.05 was considered significant.
 M. Ito et al. / En6ironmental Toxicology and Pharmacology 9 (2001) 71–78 73

3. Results

3.1. Changes in serum glucose, insulin and total

cholesterol le6els after streptozotocin administration

The non-fasting serum glucose levels of the normal

group were about 200 mg/dl throughout the initial
observation period of 1 – 9 weeks (Fig. 1(A)). In the
groups administered 150 and 200 mg/kg STZ, the
serum glucose levels rose markedly from 1 week after
STZ administration and the high glucose levels were
maintained for 9 weeks (150 mg/kg STZ: 500 –700
mg/dl; 200 mg/kg STZ: 700 – 800 mg/dl) (Ito et al.,
1999). The glucose levels of the group administered 100
mg/kg STZ were within a normal range at 1 week after
STZ administration, but rose thereafter till 9 weeks (1
week: 266958 mg/dl; 9 weeks: 682956 mg/dl) (Ito et
al., 1999). However, in the mice administered 75 mg/kg
STZ, a rise of glucose levels was not observable even at
9 weeks.
In the groups administered 100 and 200 mg/kg STZ
and in the normal group, the serum glucose levels were
measured again at 12 and 24 weeks. In the groups
administered 100 and 200 mg/kg STZ, the glucose levels
at 12 and 24 weeks were approximately the same as Fig. 2. Changes in serum glucose and insulin levels in mice within 72
h after a single i.p. injection of (A) 100 mg/kg or (B) 200 mg/kg STZ.
Each plot denotes the mean 9SE for eight mice.

those at 9 weeks in both groups. The glucose levels of

the normal group were about 200 mg/dl in both weeks.
The non-fasting serum insulin levels were determined
only in the groups administered 100 and 200 mg/kg
STZ and in the normal groups (Fig. 1(B)). The insulin
levels of the mice administered 100 mg/kg STZ were
normal from 1 week until 24 weeks when the experi-
ment terminated (1–9 weeks; Ito et al., 1999). On the
other hand, the insulin levels of the group administered
200 mg/kg STZ were below measurable values (1–9
weeks; Ito et al., 1999).
Only in the group administered 200 mg/kg STZ, the
non-fasting serum total cholesterol levels were signifi-
cantly higher than those of the normal group (Fig.
1(C)). However, the total cholesterol levels of the mice
administered 75, 100 and 150 mg/kg STZ were within
the normal range.

3.2. Changes in serum glucose and insulin le6els within

72 h after streptozotocin administration

In the group administered 100 mg/kg STZ, a tran-

sient rise in serum glucose levels was observed at 2 h
Fig. 1. Changes in (A) non-fasting serum glucose, (B) insulin and (C) after STZ administration (Fig. 2(A)). All mice given
total cholesterol levels in mice after a single i.p. injection of various
100 mg/kg STZ returned to normoglycemia within 24 h.
doses of STZ. The data obtained for 1–9 weeks after 100, 150 and
200 mg/kg STZ administration have been already reported by us (Ito On the other hand, in the group administered 200
et al., 1999). Each plot denotes the mean 9SE for 8–10 mice. mg/kg STZ, a transient rise in the serum glucose levels
Significantly different from respective normal, **PB 0.01. was observed at 2 h after STZ administration and
74 M. Ito et al. / En6ironmental Toxicology and Pharmacology 9 (2001) 71–78

returned to near normal levels by 7 h (Fig. 2(B)). The urine volume per 24 h in mice of the 150 and 200
Thereafter, the glucose levels of group administered 200 mg/kg STZ groups was markedly greater than that of
mg/kg STZ again rose and stable hyperglycemia was normal group throughout the period of 3–9 weeks (Fig.
observed at 24 and 72 h. 3(B)). In addition, the drinking water and food con-
Serum insulin levels for the 100 and 200 mg/kg STZ sumption per 24 h of the 150 and 200 mg/kg STZ
groups at 7 h after STZ administration were 13.6 and groups was also markedly greater than that of normal
14.6 times higher, respectively, than the levels at 20 h group throughout the same period (Fig. 3(C) and (D)).
before STZ administration (Fig. 2(A) and (B)). There- On the other hand, the urine volume, and the drinking
after, in the 100 mg/kg STZ group, the insulin levels water and food consumption in the 100 mg/kg STZ
slowly fell but the levels at 72 h still remained at least group steadily increased from 1–9 weeks but the degree
twice as high as the levels at 20 h before STZ adminis- of increases was less than that found in the 150 or 200
tration. In the 200 mg/kg STZ group, the insulin levels mg/kg STZ group even at 9 weeks. These three parame-
rapidly fell and were under measurable values at 48 and ters in the 75 mg/kg STZ group were similar to those in
72 h. the normal group for this period. At 24 weeks, the
urine volume, and the drinking water and food con-
3.3. Changes in body weight, urine 6olume, and sumption in 200 mg/kg STZ group had markedly de-
drinking water and food consumption after creased from the levels observed at 9 weeks, but these
streptozotocin administration parameters in the 100 mg/kg STZ group remained at
the same level as those at 9 weeks.
The body weights of mice in the 200 mg/kg STZ
group significantly decreased throughout the 9-week- 3.4. Serum glucose and insulin responses to oral
observation, compared to those before STZ administra- glucose loading
tion, while the weights of the 150 mg/kg STZ group did
not change for this period (Fig. 3(A)). The body Glucose tolerance test in the 100 mg/kg STZ and
weights of mice in the 75 and 100 mg/kg STZ groups normal mice was carried out at various intervals after
increased with the passage of time; no significant differ- STZ administration. The fasting serum glucose levels of
ence was observed between the two groups and the the 100 mg/kg STZ group at 0.5 h before oral glucose
normal group. The body weights of all animals in the loading had already been significantly higher than those
100 mg/kg STZ group thereafter further continued to of normal group at 3 days after STZ administration
increase until 24 weeks and was not significantly differ- (Fig. 4(A)). Thus, in the 100 mg/kg STZ group, hyper-
ent from that of normal group. glycemia was apparent even when the mice were fasted.
Thereafter, the fasting glucose levels before glucose
loading of the STZ group gradually rose as time went
on. In both groups, serum glucose levels after oral
glucose loading reached a peak at 0.5 h and then
gradually decreased. After glucose loading, the serum
glucose levels of the 100 mg/kg STZ group were signifi-
cantly higher than those of the normal group at all
points of measurement at 2 weeks (Fig. 4(A)). There-
after, the degree of glucose tolerance gradually reduced
as time went on.
The fasting serum insulin levels of the 100 mg/kg
STZ group at 0.5 h before oral glucose loading were
significantly higher than those of the normal group only
at 3 days after STZ administration (Fig. 4(B)). The
serum insulin level of the normal group after oral
glucose loading was markedly higher at each point of
measurement. The degree of the rise of the insulin levels
of the 100 mg/kg STZ group after glucose loading was
similar to that of the normal group up to 1 week.
Fig. 3. Changes in (A) body weight, (B) urine volume and (C) However, the insulin response to glucose stimulation in
drinking water and (D) food consumption in mice after a single i.p.
injection of various doses of STZ. Each plot denotes the mean 9SE
the STZ group was less, but not significantly, than that
for 8– 10 mice. Significantly different from respective normal, *PB of normal group at 2 weeks and was completely lost by
0.05, **PB0.01. 12 weeks.
 M. Ito et al. / En6ironmental Toxicology and Pharmacology 9 (2001) 71–78 75

Fig. 4. Changes in (A) serum glucose and (B) insulin responses to oral glucose loading (2 g/kg) in mice after a single i.p. injection of 100 mg/kg
STZ. Blood samples were collected at 0.5 h (0 h in figure) before and at 0.5, 1, 2 and 4 h after glucose loading for determination of serum glucose.
Each plot denotes the mean 9 SE for eight mice. Significantly different from respective normal, *PB 0.05, **PB0.01.

3.5. Islet morphology decreased till 4 h when the experiment terminated (Fig.
7(A) and (B)). In the 100 mg/kg STZ mice, tolbutamide
There was a marked involution of pancreatic islets in (300 mg/kg, p.o.) produced a significant decrease in the
the 200 mg/kg STZ mice, and the area markedly de- glucose levels, compared to the control at 0.5, 1 and 2
creased from 1 week after administration (Fig. 5(A)). h after glucose loading (Fig. 7(A)). In the normal mice,
On the other hand, in the 100 mg/kg STZ mice, the this drug (100 mg/kg, p.o.) also significantly decreased
area of islets was normal at 1 week and then decreased the glucose levels at 0.5, 1 and 2 h. However, in the 200
gradually as time went on (Fig. 5(A) and (B)). mg/kg STZ mice, this drug (300 mg/kg, p.o.) was
When islet glucagon (a-cells)- and insulin-immunore- ineffective in decreasing the glucose levels.
active cells (b-cells) were observed, there was a marked In the 100 mg/kg STZ mice, troglitazone (100 mg/kg,
reduction in the number of b-cells from 1 week in the p.o.) caused a marked serum glucose-decreasing effect
200 mg/kg STZ mice (Fig. 5(A), (B) and Fig. 6). In 100 throughout the period of 0.5 –4 h after glucose loading
mg/kg STZ mice, the number of b-cells was almost (Fig. 7(B)). However, in the normal and 200 mg/kg
normal at 1 week after administration and then contin- STZ mice, this agent (100 mg/kg, p.o.) failed to de-
ued to reduce till 24 weeks (Fig. 6). In contrast, there crease the glucose levels at all hours measured.
was a relative increase in the number of a-cells in the
100 mg/kg STZ mice.
4. Discussion
3.6. Effects of tolbutamide and troglitazone on glucose
tolerance STZ has been commonly used to induce not only
animal models of IDDM (Rakieten et al., 1963; Brod-
After oral glucose loading, serum glucose levels of all sky and Logothetopoules, 1969; Junod et al., 1969;
groups reached a peak at 0.5 h and then gradually Rerup, 1970), but also NIDDM with hypoinsulinemia
76 M. Ito et al. / En6ironmental Toxicology and Pharmacology 9 (2001) 71–78

Fig. 5. Insulin-immunostaining of a- and b-cells in the islets of pancreas in normal, 100 and 200 mg/kg STZ-treated mice at day 1, (A) 1 and 4
weeks, (B) 8, 12 and 24 weeks after a single i.p. injection of STZ. Original magnification: 100 ×.

by neonatal (1- or 2-day-old mice or rats) STZ marked involution of pancreatic islets and a marked
administration (Giroix et al., 1983; Tsuji et al., 1988; reduction in the number of islet insulin-immunoreactive
Wang et al., 1994). It has been reported that STZ is cells (b-cells) from 1 week after administration. On the
capable of producing mild to severe types of diabetes other hand, the area of islets and the number of b-cells
according to the dosage used, when it is given to adult in 100 mg/kg STZ mice continued to reduce gradually
rats by either single i.v. or i.p. injection (Junod et al., as time went on throughout the 24-week-observation
1969). period after administration. The exact mechanism of
In the previous and present studies, when STZ was the decrease in the number of b-cells during the pro-
i.p. injected in 8-week-old ICR mice at doses ranging gression of this diabetes is still unclear. It is of interest,
from 75 to 200 mg/kg, only 100 mg/kg STZ induced however, that in 100 mg/kg STZ mice, non-fasting
slowly-progressive diabetes mellitus. In other words, in
the mice administered 100 mg/kg STZ, non-fasting
serum glucose levels continued to increase gradually
after STZ administration without affecting the
non-fasting serum insulin levels. In contrast to the 100
mg/kg STZ mice, the serum glucose levels of 200 mg/kg
STZ mice sharply rose with an accompanying marked
fall in serum insulin levels from the first day after STZ
administration. These results strongly suggest that 100
mg/g STZ-induced diabetic mouse model is non-insulin
dependent, while the 200 mg/kg STZ-induced one is
insulin-dependent.
Fig. 6. Changes in the percentage of the number of glucagon (a-cells)-
In the present study, morphological observation in and insulin-immunoreactive cells (b-cells) in the islets of pancreata in
the mice administered 200 mg/kg STZ revealed a mice after a single i.p. injection of 100 or 200 mg/kg STZ.
 M. Ito et al. / En6ironmental Toxicology and Pharmacology 9 (2001) 71–78 77

Skoglund, 1989) or rats (Tsuji et al., 1988). Thus, the

insulin response to glucose loading in 100 mg/kg STZ-
induced diabetic mouse model was similar to that in
neonatal STZ-induced diabetic mouse or rat model.
It is conceivable that the impairment of glucose-stim-
ulated insulin secretion in 100 mg/kg STZ-induced dia-
betic mouse model may be in part related to the
decrease in the number of islet b-cells. As it has been
reported that hyperglycemia (Robertson et al., 1992;
Moran et al., 1997) or hyperlipidemia, (Zhou and Grill,
1994) impairs b-cell functions such as the inhibition of
glucose-stimulated insulin secretion or islet glucose
metabolism, it may be that the abnormalities of glucose
or lipid metabolism further promote the impairment of
the function and proliferative capacity of b-cells.
In the present experiment, in order to further clarify
characteristics of this progressive diabetic model, we
examined the effects of tolbutamide, a first generation
Fig. 7. Effects of (A) tolbutamide and (B) troglitazone on glucose
tolerance tests in normal mice and mice administered 100 or 200 sulfonylurea, which promotes insulin secretion, and
mg/kg STZ at 12 weeks after a single i.p. injection of STZ. Blood troglitazone, an antidiabetic agent, which reduces pe-
samples were collected at 0.5 h (0 h in figure) before and at 0.5, 1, 2 ripheral insulin resistance, on glucose tolerance in 100
and 4 h after glucose loading for determination of serum glucose. mg/kg STZ mice in comparison with normal and 200
Each plot denotes the mean 9SE for six mice. Significantly different
mg/kg STZ mice.
from respective normal, *P B 0.05, **PB0.01.
It has been reported that tolbutamide has blood
glucose-lowering action in normal or diabetic rats with
serum insulin levels were maintained at normal levels
b-cells which can secrete insulin, but this agent does not
throughout the observation period, although there was
act in insulin-deficient diabetes (Saxena et al., 1996). In
a time-course-related reduction in the number of b-
our experiment, tolbutamide significantly lowered
cells. These results suggest that, in 100 mg/kg STZ
serum glucose levels after oral glucose loading in nor-
mice, the remaining b-cells escaping from the attack of
mal (100 mg/kg, p.o.) and 100 mg STZ mice (300
STZ may cause an oversecretion of insulin to maintain mg/kg, p.o.). However, in 200 mg/kg STZ mice which
normoglycemia. Therefore, it is reasonable to assume are insulin-dependent, this agent failed to lower the
that the reduction in the number of b-cells may be glucose levels. These results suggest that tolbutamide
attributed to the inhibition of their proliferation due to has a hypoglycemic action in normal and 100 mg/kg
the excess activity of the remaining b-cells. These results STZ animals which still have the ability to secrete
also suggest that this slowly progressive NIDDM insulin from b-cells, while it is ineffective in insulin-de-
mouse model may convert into IDDM after long-term pendent diabetic animals administered 200 mg/kg STZ.
chronic hyperglycemia. Troglitazone (100 mg/kg, p.o.) was significantly effec-
In the second experiment, we carried out glucose tive in lowering the glucose levels in 100 mg/kg STZ
tolerance tests at various intervals after STZ adminis- animals. However, in normal and 200 mg/kg STZ
tration to further clarify the characteristics of this 100 animals, this agent did not exhibit hypoglycemic action.
mg/kg STZ-induced diabetic mouse model. The serum Fujiwara et al. (1988) have reported that troglitazone
glucose levels of these diabetic mice after glucose load- (CS-045) is effective in insulin-resistant diabetic animal
ing were significantly higher than those of normal mice models such as the KK mouse, the ob/ob mouse and
at 2 weeks after STZ administration, and thereafter the the Zucker fatty rat. They also have shown that this
degree of glucose tolerance gradually reduced as time agent is ineffective in STZ-induced insulin-dependent or
went on. In addition, the insulin response to glucose normal animals. Therefore, in the present study, it is
loading in the STZ mice became weaker in direct postulated from the results obtained with troglitazone
proportion to the reduction in glucose tolerance and that insulin resistance is developed in 100 mg/kg STZ-
was completely lost by 12 weeks. Therefore, the results induced diabetic mice (12 weeks after STZ treatment),
obtained from this test indicate that glucose-stimulated although hyperinsulinemia was not seen. However, fur-
insulin secretion is markedly impaired in the 100 mg/kg ther experiments are necessary to clarify whether or not
STZ-induced diabetic mouse model. insulin resistance is developed in this model.
It has been also demonstrated that insulin responses Whether or not slowly progressive diabetes mellitus is
to glucose loading are impaired during the progression induced in mice of other strains or animals of different
of diabetes in STZ-injected neonatal mice (Ahrén and species by a single i.p. injection of subdiabetogenic dose
78 M. Ito et al. / En6ironmental Toxicology and Pharmacology 9 (2001) 71–78

of STZ remains unclear. It has been shown that there is Herr, R.R., Eble, T.E., Bergy, M.E., Jahnke, H.K., 1959. Isolation and
characterization of streptozotocin. Antibiot. Ann. 60, 236–240.
a temporary rise of blood glucose levels followed by
Ito, M., Kondo, Y., Nakatani, A., Naruse, A., 1999. New model of
spontaneous recovery within a week, when a small dose progressive non-insulin-dependent diabetes mellitus in mice in-
(30 or 40 mg/kg, i.v.) of STZ was injected in non-fasted duced by streptozotocin. Biol. Pharm. Bull. 22, 988– 989.
male Sprague –Dawley rats weighing approximately 250 Junod, A., Lambert, A.E., Orci, L., Pictet, R., Gonet, A.E., Renold,
g (Ar’rajab and Ahrén, 1993). It is of interest that in A.E., 1967. Studies of the diabetogenic action of streptozotocin.
Proc. Soc. Exp. Biol. Med. 126, 201– 205.
adult ICR mice, unlike adult rats, progressive diabetes Junod, A., Lambert, A.E., Stauffacher, W., Renold, A.E., 1969.
mellitus can be induced by the administration of small Diabetogenic action of streptozotocin: relationship of dose to
dose of STZ. metabolic response. J. Clin. Invest. 48, 2129– 2139.
In summary, the present study indicates that the Moran, A., Zhang, H.-J., Olson, L.K., Harmon, J.S., Poitout, V.,
Robertson, R.P., 1997. Differentiation of glucose toxicity from beta
progressive diabetic mouse model induced by a single
cell exhaustion during the evolution of defective insulin gene
i.p. injection of 100 mg/kg STZ is NIDDM, which is expression in the pancreatic islet cell line, HIT-T15. J. Clin. Invest.
characterized by the impaired insulin response to glu- 99, 534– 539.
cose stimulation. In addition, this new diabetic model Rakieten, N., Rakieten, M.L., Nadkarni, V., 1963. Studies on the
may be useful to clarify the mechanism for the progres- diabetogenic action of streptozotocin (NSC-37917). Cancer
Chemother. Rep. 29, 91 – 98.
sion of NIDDM in human and to screen antidiabetic Rerup, C.C., 1970. Drugs producing diabetes through damage of
substances. insulin secreting cells. Pharmacol. Rev. 22, 485– 518.
Robertson, R.P., Zhang, H.-J., Pyzdrowski, K.L., Walseth, T.F., 1992.
Preservation of insulin mRNA levels and insulin secretion in HIT
cells by avoidance of chronic exposure to high glucose concentra-
References
tions. J. Clin. Invest. 90, 320– 325.
Saxena, A.M., Murthy, P.S.R., Mukherjee, S.K., 1996. Mode of action
Ahrén, B., Skoglund, G., 1989. Insulin secretion in neonatally strepto- of three structurally different hypoglycemic agents: a comparative
zotocin-injected mice. Diabetes Res. 11, 185–190. study. Ind. J. Exp. Biol. 34, 351– 355.
Ar’rajab, A., Ahrén, B., 1993. Long-term diabetogenic effect of Tsuji, K., Taminato, T., Usami, M., Ishida, H., Kitano, N., Fukumoto,
streptozotocin in rats. Pancreas 8, 50–57. H., Koh, G., Kurose, T., Yamada, Y., Yano, H., Seino, Y., Imura,
Brodsky, G., Logothetopoules, J., 1969. Streptozotocin induced dia- H., 1988. Characteristic features of insulin secretion in the strepto-
betes in the mouse and guinea pig. Diabetes 18, 606–611. zotocin-induced NIDDM rat model. Metabolism 37, 1040–1044.
Fujiwara, T., Yoshioka, S., Yoshioka, T., Ushiyama, I., Horikoshi, H., Wang, R.N., Bouwens, L., Kloppel, G., 1994. Beta-cell proliferation
1988. Characterization of new oral antidiabetic agent CS-045. in normal and streptozotocin-treated newborn rats; site, dynamics
Diabetes 37, 1549– 1558. and capacity. Diabetologia 37, 1088– 1099.
Giroix, M.-H., Portha, B., Kergoat, M., Bailbe, D., Picon, L., 1983. Zhou, Y.-P., Grill, V.E., 1994. Long-term exposure of rat pancreatic
Glucose insensitivity and amino-acid hypersensitivity of insulin islets fatty acids inhibits glucose-induced insulin secretion and
release in rats with non-insulin-dependent diabetes. Diabetes 32, biosynthesis through a glucose fatty acid cycle. J. Clin. Invest. 93,
445– 451. 870– 876.