Life Sciences 286 (2021) 120062

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Life Sciences
journal homepage: www.elsevier.com/locate/lifescie

A new mouse model of type 2 diabetes mellitus established through

combination of high-fat diet, streptozotocin and glucocorticoid
Sha Liu a, Lingling Ma a, Xiaoyu Ren a, Wenling Zhang b, Dian Shi a, Yanbei Huo b, Yupei Ba b,
Yana Bai b, Ning Cheng a, *
a
Key Laboratory of Preclinical Study for New Drugs of Gansu Province, Basic Medical College, Lanzhou University, Lanzhou, Gansu, PR China
b
Institute of Epidemiology and Statistics, School of Public Health, Lanzhou University, Lanzhou, Gansu, PR China

A R T I C L E I N F O A B S T R A C T

Keywords: Aim: A stable induced type 2 diabetes model (T2DM) still needs to be explored for basic and clinical research, due
T2DM model to nonuniform model methods and unstable consequences. Our aims were to explore and establish an optimized
Insulin resistance induced T2DM model in mice that exhibits insulin resistance and β-cell damage.
Glucocorticoid
Materials and methods: C57BL/6 mice were treated with a high-fat diet (HFD), streptozotocin (STZ) and dexa­
β-Cell damage
methasone (DEX) at different doses and in combination. The general growth status, blood glucose and fasting
Streptozotocin
insulin were detected, and the success rate and insulin sensitivity indices were calculated.
Key finding: Low-dose STZ injection multiple times was more secure in the process of T2DM model production.
Combined intervention was more efficient in reducing insulin sensitivity and improving the success rate of T2DM
model construction.
Significance: Combined with a high-fat diet, glucocorticoids and streptozotocin, a new mouse model of T2DM
with insulin resistance and β-cell damage could be established. The optimized experimental method can serve as
a stable model for further studies on the mechanisms and therapy of T2DM.

1. Introduction
Type 2 diabetes mellitus (T2DM) is a metabolic syndrome with
persistent hyperglycemia [1]. Persistent high blood glucose and insulin
resistance are the main characteristics of diabetes. At present, commonly
used animal models of diabetes include two categories. One is the
spontaneous animal model caused by genetic defects, such as ob/ob
mice, db/db mice and KK mice [2], and the other is the nonspontaneous
animal model induced by human factors. Compared with the sponta­
neous model, the induced model is more popular and has more advan­
tages, including low cost, stable effect and easy acquisition.
In terms of the induced model, high-fat diet (HFD) induction can
mimic the development of insulin resistance and hyperglycemia caused
by diet with simple operation. However, it always takes a long time to
feed and detect changes in blood glucose and insulin frequently.
Chemical induction is a good method to induce hyperglycemia by
damaging β-cells specifically, but the intervention dose is difficult to
control, and it easily causes type 1 diabetes mellitus (T1DM) and even
death. It is also not an appropriate inducer in terms of inducing insulin
resistance in animals. Currently, HFD combined with streptozotocin
(STZ) is one of the prevalent methods of inducing T2DM in animal
models and can easily show the characteristics of diabetes mellitus [3].
However, there are no specific standards for the intervention time of
HFD and the treatment dose of STZ in this model; moreover, this method
is commonly accompanied by unstable consequences.
At present, the clinical treatment of T2DM mainly focuses on glucose
control, and the improvement of insulin resistance and prevention of
diabetic complications are less executed [4]. However, a T2DM mouse
model, which is consistent with the pathogenesis of T2DM and has a
stable effect on insulin resistance and β-cell damage, is scarce. Gluco­
corticoids have powerful anti-inflammatory, anti-immune and anti-
shock effects and are widely used in the clinical treatment of immune
diseases, blood diseases, organ rejection and other diseases. In addition,
unreasonable dosages also affect the metabolism of lipids and glucose

Abbreviations: T1DM, type 1 diabetes mellitus; T2DM, type 2 diabetes mellitus; STZ, streptozotocin; DEX, dexamethasone; HOMA, homeostasis model assessment;
HFD, high-fat diet; GLUT2, glucose transporter-2; IR, insulin resistance; QUICKI, quantitative insulin sensitivity check index; Fins, fasting insulin.
* Corresponding author at: Key Laboratory of Preclinical Study for New Drugs of Gansu Province, Basic Medical College, Lanzhou University, Lanzhou, Gansu
730000, PR China.
E-mail address: chengn@lzu.edu.cn (N. Cheng).

https://doi.org/10.1016/j.lfs.2021.120062
Received 5 May 2021; Received in revised form 11 October 2021; Accepted 13 October 2021
Available online 18 October 2021
0024-3205/© 2021 Published by Elsevier Inc.
S. Liu et al.
and even cause metabolic syndrome, such as Cushing's syndrome char­
acterized by centripetal obesity, hyperlipidemia, hypertension and in­
sulin resistance [5–7]. By adding glucocorticoids to the HFD + STZ
model and optimized design, we established a mouse model of T2DM,
which has a stable effect on insulin resistance and hyperglycemia and
improved repeatability and success rate.
2. Materials and methods
2.1. Animals
6–8 weeks old male C57BL/6 mice (purchased from Lanzhou Vet­
erinary Research Institute, Chinese Academy of Agricultural Sciences,
Animal Production License No.: [SCXK (Gan) 2015-0001]) were kept in
the Animal Experiment Center of Lanzhou University [SYXK (Gan)
2018-0002], with the temperature of 20–25 ◦ C and humidity of 50 ±
10%. During that period, water and feed were free to take in the light/
dark cycle of 12 h every day.
2.2. Reagents and instruments
HFD: 20% sucrose, 10% lard, 2.5% cholesterol, 1% sodium cholate,
1% mineral mixture and 0.5% vitamin mixture is added on the basis of
65% mouse maintenance diet (Beijing Boai Port Trading Co., LTD.).
Dexamethasone sodium phosphate injection (DEX, 5 mg/ml, Sinophar­
ynx Rongsheng Pharmaceutical Co., LTD., H41020036). STZ (Sigma-
Aldrich, S0130). Citric acid buffer (0.1 mM, pH = 4.5). Acidity meter
(OHAUS STARTER 3100). Electronic balance (Sartorius, BSA224S)
2.3. Experimental procedures
First, we performed an experiment to observe the long-term stability
of the combined intervention model. Thirty mice were randomly divided
into 3 groups: normal control group (NC, n = 10), combined interven­
tion Group a (HFD + DEX + STZ-a, STZ 50 mg/kg, injection 5 times, n =
10), and combined intervention Group b (HFD + DEX + STZ-b, STZ 110
mg/kg, injection one time, n = 10). The NC group was fed a normal diet,
while the other groups were fed a HFD until the end of the experiment.
After 4 weeks of dietary induction, HFD + DEX + STZ-a was injected
with DEX (1 mg/kg/day, subcutaneous injection) 7 times [8,9], followed
by STZ (50 mg/kg/day, intraperitoneal injection) 5 times. HFD + DEX +
STZ-b was injected with DEX (1 mg/kg/day, subcutaneous injection) 7
times continuously, followed by STZ (110 mg/kg, intraperitoneal in­
jection) 1 time. The STZ injection was prepared with fresh precooled
citric acid buffer before use. The NC was given a corresponding dose of
buffer solution at the same time. After the intervention, each group was
observed for 6 weeks (Fig. 1).
After that, we compared the effect of methods through experiments.
The mice were randomly divided into four groups: normal control group
(NC, n = 10), model control group (MC, n = 30), combined intervention
Group 1 (HFD + DEX + STZ-1, STZ 50 mg/kg, injection 5 times, n = 30),
and combined intervention Group 2 (HFD + DEX + STZ-2, STZ 110 mg/
kg, injection one time, n = 30). The dietary intervention was the same as
the previous experiment. After 4 weeks of dietary induction, the MC
group was injected with STZ (50 mg/kg/day, intraperitoneal injection) 5
Fig. 1. Design of the T2DM mouse model to observe long-term stability.
2
Life Sciences 286 (2021) 120062
times continuously. The HFD + DEX + STZ-1 and HFD + DEX + STZ-2
groups were the same as the HFD + DEX + STZ-a and HFD + DEX + STZ-
b groups. In our previous experiments, we found that the mouse model
with the combined intervention could be maintained stably for 6 weeks
at least. To optimize the condition of modeling, each group was
observed for 12 days after intervention (Fig. 2).
In the above experiment, HFD + DEX + STZ-1 group had a more
stable effect on hyperglycemia and insulin resistance. For dosage opti­
mization detection, 80 mice were randomly divided into two groups: the
combined intervention group with 50 mg/kg STZ (HFD + DEX + STZ-
50, STZ 50 mg/kg, injection 5 times, n = 30) and the combined inter­
vention group with 60 mg/kg STZ (HFD + DEX + STZ-60, STZ 60 mg/
kg, injection 5 times, n = 30). The mice were fed a HFD for 4 weeks and
then injected with DEX (1 mg/kg/day, subcutaneous injection) 7 times
continuously. Subsequently, the HFD + DEX + STZ-50 and HFD + DEX
+ STZ-60 groups were treated with STZ 5 times at 50 mg/kg and 60 mg/
kg doses, respectively. During the experiment, the general growth status
of animals in groups was observed.
Determination of blood glucose: In the experiment that evaluating
the model stability, blood glucose was measured weekly. In the exper­
iment of optimizing method and dosage, blood glucose was measured on
the 3rd, 5th and 12th days of the observation period. After fasting for 2 h
in the morning, we took blood from the tail tip of the mice with a blood
collection needle, and the blood glucose was measured with a Roche
Glucometer (ACCU-Chek® Performa) and Roche blood glucose test
paper (ACCU-Chek® Performa). If the value was higher than 11.1
mmol/L, it was considered to be a successful sample. If it was greater
than 20 mmol/L, it was considered too high and may become T1DM. The
success rate was calculated after the experiment. An oral glucose toler­
ance test (OGTT) was performed after observation period (the 13th day).
The mice in each group were fasted for 14 h with free water and new
pads. Then, fresh glucose solution (2 g/kg of body weight) was given by
gavage. Blood glucose was measured at 0 h, 0.5 h, 1 h and 2 h with a
Roche Glucometer. After that, the blood glucose-time curve was plotted,
and the area under the curve (AUC) was calculated.
Evaluation of insulin sensitivity: Blood samples taken from the retro-
orbital region after fasting overnight, centrifugation to separate the
serum after measuring fasting blood glucose (the 14th day). Fasting
insulin (Fins) content was detected by ELISA kit (Wuhan Huamei
Biotechnology Co., LTD., CSB-E05071 m). The quantitative insulin
sensitivity check index QUICKI {=1 / [log(Fins, μU/ml) + log(FPG, mg/
dl)]} and HOMA value, including HOMA-IR (=FPG × FINS / 22.5),
HOMA-IS (=1 / HOMA-IR) and HOMA-β [=20 × FINS / (FPG − 3.5)%]
[10], were calculated after the experiment.
2.4. Statistics
SPSS 22.0 (IBM, Armonk, NY, USA) and Origin 2017 (OriginLab,
USA) statistical software were used to analyze the results. Descriptive
statistics are expressed as the mean ± SD (M ± S), and categorical
variables are expressed as frequencies (percentages). Continuous
Fig. 2. Design of optimization method of combined intervention method for
T2DM mouse model.
S. Liu et al.
variables were compared using Student's t-test between 2 groups, anal­
ysis of variance (ANOVA) was used for comparison of more than two
groups, and LSD was used for post hoc test. Categorical variables were
compared using the chi-square test. p < 0.05 was considered statistically
significant.
3. Results
3.1. The stability of combined intervention model
3.1.1. The growth status and blood glucose
The results showed that within six weeks after intervention, the body
weight of mice in the combined intervention groups decreased obviously
and was significantly lower than that in the NC group (Fig. 3a, p < 0.05).
The body weight of HFD + DEX + STZ-a group remained stable for six
weeks with no significant fluctuations. The body weight of HFD + DEX
+ STZ-b group decreased among the first three weeks and kept stable
after fluctuation, which remained lower than the other groups (p <
0.05). The blood glucose levels of the HFD + DEX + STZ-a and HFD +
DEX + STZ-b groups were always significantly higher than NC group
(Fig. 3b, p < 0.05). It was obviously that the blood glucose peak in HFD
+ DEX + STZ-a reached in the 3rd week and then remained stable, while
the peak in HFD + DEX + STZ-b group reached in the 2nd week and then
gradually decreased.
3.1.2. Insulin sensitivity
The insulin sensitivity results showed that the mice still had signif­
icant insulin resistance at 6 weeks after the combined intervention. As
shown in Fig. 4a, the OGTT curves of the HFD + DEX + STZ-a and HFD
+ DEX + STZ-b groups were significantly higher than those of the NC
group (p < 0.05), but there was no significant difference between the
two intervention groups. After calculating the AUC, we found that the
AUC value of the combined intervention group was significantly higher
than that of the NC group (Fig. 4b, p < 0.05). Compared with NC group,
FINS and HOMA-IR of HFD + DEX + STZ-a and HFD + DEX + STZ-b
groups were significantly increased (Fig. 4c, d, p < 0.05), while HOMA-
IS, HOMA-β and QUICKI were significantly decreased (Fig. 4e, f, g, p <
0.05), but there was no significant difference between the two inter­
vention groups (p > 0.05).
3.2. Optimization the method of combined intervention T2DM mouse
model
3.2.1. The growth status
During the HFD feeding period, mice in each group had smooth fur
and active movement. Compared with the NC group, the water intake
and body weight of the other three groups showed no significantly
different (Fig. 5d, g), but the food intake was increased (Fig. 5a). During
DEX intervention, the food intake and water intake in HFD + DEX +
STZ-1 and HFD + DEX + STZ-2 were all decreased compared with the
MC (Fig. 5b, e, p < 0.05). A significant reduction (Fig. 5h, p < 0.05) in
Fig. 3. Blood glucose and body weight changes of NC (n = 10), HFD + DEX +
STZ-a (n = 10) and HFD + DEX + STZ-b groups (n = 10). a, Body weight
changes in each group; b, Blood glucose changes in each group. *p < 0.05
compared with NC, &p < 0.05 compared with HFD + DEX + STZ-a.
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Life Sciences 286 (2021) 120062
Fig. 4. Insulin sensitivity indicators of NC (n = 10), HFD + DEX + STZ-a (n =
10) and HFD + DEX + STZ-b (n = 10) groups. a, OGTT in each group; b, AUC in
each group; c, Fins (mIU/L) in each group; d, HOMA-IR in each group s; e,
HOMA-IS value in each group; f, HOMA-β value in each group; g, QUICKI value
AUC in each group. *p < 0.05 compared with NC group.
body weight was observed in the HFD + DEX + STZ-1 and HFD + DEX +
STZ-2 groups compared with the NC and MC groups after DEX inter­
vention. After STZ treatment, the food and water intake in the MC, HFD
+ DEX + STZ-1 and HFD + DEX + STZ-2 groups was significantly
increased compared with that in the NC group (Fig. 5c, f, p < 0.05), but
body weight was significantly decreased (Fig. 5i, p < 0.05). The fur of
mice appeared dim and fell off, and the amount of exercise reduced.
3.2.2. Blood glucose
We measured the blood glucose of mice on the 3rd, 5th, and 12th
days during the observation period. In contrast to the NC group, the
blood glucose of other groups increased significantly (Fig. 6a, b, c). The
results show that blood glucose of the MC group increased clearly, from
11.7 mmol/L on the 3rd day to 15.0 mmol/L on the 5th day and to 16.2
mmol/L on the 12th day. The blood glucose of HFD + DEX + STZ-1 also
showed a steady rise, from 11.1 mmol/L on the 3rd day to 12.9 mmol/L
on the 5th day to 16.6 mmol/L on the 12th day. Compared with the MC
group, the blood glucose of the HFD + DEX + STZ-1 group increased
slowly, but the difference was not statistically significant (p > 0.05). On
the other hand, the blood glucose of HFD + DEX + STZ-2 mice first
increased and then decreased. On the 3rd day, it was lower than the MC
group and HFD + DEX + STZ-1, but it increased rapidly on the 5th day
and higher than the other groups. However, it decreased on the 12th day
and was lower than the MC group and HFD + DEX + STZ-1 again. In
general, blood glucose in HFD + DEX + STZ-1 and HFD + DEX + STZ-2
remained stable during the observation period, and there was no sig­
nificant difference when compared with MC (p > 0.05).
3.2.3. Insulin sensitivity
The results showed that the insulin sensitivity of mice declined
significantly after treatment with DEX, as reported previously [11,12].
The OGTT and AUC of HFD + DEX + STZ-1 and HFD + DEX + STZ-2
groups were significantly higher than MC and NC group (Fig. 7a, b, p <
0.05), and the data of HFD + DEX + STZ-1 group was higher than HFD
+ DEX + STZ-2 group (p > 0.05). In contrast to the MC and NC groups,
the Fins levels of HFD + DEX + STZ-1 and HFD + DEX + STZ-2 groups
elevated significantly (Fig. 7c, p < 0.05). Meanwhile, the change of
QUICKI and HOMA values were also evident. After combined treatment,
QUICKI and HOMA-IS values of mice declined significantly (Fig. 7g, e, p
< 0.05), while the HOMA-IR value increased obviously (Fig. 7f, p <
0.05). The increase was particularly noticeable in HFD + DEX + STZ-1
group. The HOMA-β levels of the HFD + DEX + STZ-1 and HFD + DEX +
STZ-2 groups declined markedly compared with that of the NC group,
but there was no significant difference from that of the MC group
(Fig. 7f, p < 0.05).
3.2.4. Success rate of modeling
After the experiment, we calculated the success rate of modeling in
each group (Fig. 8). In contrast, the results of model establishment in the
S. Liu et al. Life Sciences 286 (2021) 120062

Fig. 5. Food intake, water intake and body weight changes of NC (n = 10), MC (n = 30), HFD + DEX + STZ-1 (n = 30) and HFD + DEX + STZ-2 (n = 30) groups
during the experiment. a, Food intake in each group during diet intervention; b, food intake in each group during DEX intervention; c, food intake of in each group
during the observation period (12 days); d, water in each group during diet intervention; e, water intake of in each group during DEX intervention; f, water intake of
in each group during the observation period (12 days); g, body weight in each group after diet intervention; h body weight in each group during DEX intervention; i,
body weight in each group after STZ intervention.

the 3rd day during the observation period (Fig. 9a, b, c). However, the
results of modeling success rate had some changes. In contrast to HFD +
DEX + STZ-50, the success rate of HFD + DEX + STZ-60 was signifi­
cantly higher (Fig. 9d, p < 0.05). The mortality rate in both groups was
0. This result indicated that 60 mg/kg STZ was better than 50 mg/kg in
establishing a T2DM mouse model.

Fig. 6. Blood glucose changes of NC (n = 10), MC (n = 30), HFD + DEX + STZ-
1 (n = 30) and HFD + DEX + STZ-2 (n = 30) groups during the observation
period (12 days). a, Blood glucose in each group on the 3rd day; b, blood
glucose of in each group on the 5th day; c, blood glucose in each group on the
12th day. *p < 0.05 compared with NC group.
HFD + DEX + STZ-1 group were better than those in the other two
groups. The success rate of HFD + DEX + STZ-1 group was significantly
higher than that of MC (p < 0.05), and the mortality rate was signifi­
cantly lower than that of MC (p < 0.05). The success rate of HFD + DEX
+ STZ-1 group was obviously higher than that of HFD + DEX + STZ-2,
and the mortality rate was lower than that of HFD + DEX + STZ-2 (p >
0.05). It was suggested that low-dose STZ injection multiple times was
more beneficial to the modeling of T2DM mice.
3.3. Optimization dosage of combined intervention T2DM mouse model
In the dosage optimization experiment, the growth status of the mice
did not differ significantly from previous experiments. There was no
significant difference in blood glucose between the two groups except on
4. Discussion
The incidence of T2DM, as a metabolic syndrome, has a straight rise
in recent decades and is a serious threat to human health. Insulin
resistance (IR) is one of the important features of T2DM and is caused by
a combination of multiple factors. IR could reduce insulin sensitivity,
hinder insulin signal transduction, suppress glucose uptake and utili­
zation, and even lead to disorders of glucose metabolism and lipid
metabolism [13–16]. It also increases the risk of complications, such as
atherosclerosis, heart disease and stroke [17]. Long-term HFD can cause
obesity, hyperlipidemia, hyperinsulinemia and insulin resistance. Ahlke
Heydemann's study [18] showed that with prolonged HFD feeding time,
fat cells increased to store excess energy, while excess lipids slowly
deposited in various organs, causing lipid toxicity, obesity, decreased
metabolic flexibility, oxidative stress and chronic inflammation. In the
experiment of optimization the method of combined intervention
model, during dietary induction, the food intake in the MC, HFD + DEX
+ STZ-1 and HFD + DEX + STZ-2 groups was significantly higher than
that in the NC group. The insulin sensitivity index of the intervention
groups changed significantly (p < 0.05), although it could be a combi­
nation of HFD and DEX.

4
S. Liu et al.
Fig. 7. Insulin sensitivity indicators of NC (n = 10), MC (n = 30), HFD + DEX + STZ-1 (n = 30) and HFD + DEX + STZ-2 (n = 30) groups. a, OGTT in each group; b,
AUC in each group; c, Fins (mIU/L) in each group; d, HOMA-IR in each group; e, HOMA-IS value in each group; f, HOMA-β in each group; g, QUICKI value in each
group. *p < 0.05 compared with NC group. #p < 0.05 compared with MC group.
Fig. 8. The success rate of modeling in the NC (n = 10), MC (n = 30), HFD +
DEX + STZ-1 (n = 30) and HFD + DEX + STZ-2 (n = 30) groups.
STZ is an antibiotic extracted from Streptomyces achromogenes, which
is a DNA alkylating reagent with a glucosamine-nitrourea structure, in
which the glucose part transports itself to islet β-cells via the glucose
transporter GLUT2, and the nitrourea part leads to DNA alkylation,
which breaks the DNA of β-cells. Studies have shown that STZ can cause
DNA fragmentation in β-cells and insulinoma cells [19–21]. As a com­
mon drug used in animal models of diabetes, STZ can selectively damage
islet β-cells, causing experimental diabetes. The degree of damage in
β-cells is associated with the dosage and frequency of STZ. Some re­
searchers have found that a single high dosage of STZ can lead to rapid
and massive necrosis of β-cells, which easily causes T1DM, while mul­
tiple low doses can cause partial damage [22,23]. Previous researches
suggested that 50 mg/kg dosage was better for modeling of T2DM by
multiple injections with STZ at low doses [24–26]. In this study, we
found that when a single high dose of STZ (110 mg/kg) led a quick rush
of blood glucose and then decreased. When the animals were treated
with multiple small doses of STZ, their blood glucose showed a trend of
increasing gradually, which could be maintained in a stable state of
hyperglycemia. By comparing the effects of different doses, we found
that the modeling success rate was superior in 60 mg/kg, which was
more conducive to establish a T2DM mouse model.
DEX is a glucocorticoid analog that induces insulin resistance and
5
Life Sciences 286 (2021) 120062
Fig. 9. Blood glucose and success rate of HFD + DEX + STZ-50 (STZ 50 mg/kg,
injection 5 times, n = 40), and HFD + DEX + STZ-60 (STZ 60 mg/kg, injection
5 times, n = 40) during the observation period (12 days). a, Blood glucose in
each group on the 3rd day; b, blood glucose in each group on the 5th day; c,
blood glucose in each group on the 12th day. d: The success rate of in each
group. &p < 0.05 compared with HFD + DEX + STZ-50.
exacerbates the symptoms of diabetes. According to previous research,
when rodents intervene, DEX could impair insulin-induced glucose up­
take and influence the metabolism of glucose [27–31]. It could also
cause mitochondrial dysfunction, which can lead to insulin resistance
[32]; however, there was no direct effect on insulin receptors [33].
Studies of insulin resistance induced by DEX in healthy subjects showed
that short-term administration of DEX stimulated insulin secretion and
inhibits the regulation of glucose and lipid metabolism by insulin
[34,35]. Other studies have indicated that low-dose intervention of DEX
by subcutaneous injection could induce endothelial dysfunction in mice,
which led to insulin resistance with increased serum insulin concen­
tration, reduced insulin tolerance, and long-term hyperglycemia [36].
Mahesh Ghaisas's study showed that 7 days of DEX (1 mg/kg) injection
induced insulin resistance in mice [8,9], and the same dosage was used
in other studies [37–39]. IR induced by DEX has also been reported in
other studies [40,41]. In our study, DEX (1 mg/kg) intervention was
added based on the HFD + STZ model, the blood glucose in the com­
bined intervention groups (HFD + DEX + STZ-1 and HFD + DEX + STZ-
2) was slightly lower than that in the MC group, but the blood glucose
S. Liu et al.
value remained stable during the observation period, and the insulin
level was significantly higher than that in the MC group (p < 0.05).
Blood glucose in the combined intervention groups (HFD + DEX + STZ-a
and HFD + DEX + STZ-b) maintained significantly higher than that in
NC group for six weeks at least that's not too different from the stability
of HFD + STZ mouse model. Studies have shown that the HFD + STZ
model can be stable for more than 4 weeks [42,43]. In the analysis of
insulin sensitivity, we discovered that compared with NC and MC
groups, the OGTT curve of combined intervention groups shifted up­
ward, AUC increased, QUICKI value and HOMA-IS decreased signifi­
cantly (p < 0.05), and the HOMA-IR was increased obviously (p < 0.05).
The same result was found in the experiment of observing stability, The
insulin sensitivity of combined intervention groups was significantly
lower than that of NC group. In addition, there were no mice dead in the
HFD + DEX + STZ-1 group; besides, the mortality in the HFD + DEX +
STZ-2 group was lower than in the MC group. The cause of death was
speculated due to the high doses of STZ in HFD + DEX + STZ-2 group.
This result indicated that DEX might have protective effect against stress
and reduce mortality.
Compared with the NC group, blood glucose of the combined inter­
vention groups significantly increased and insulin sensitivity signifi­
cantly decreased both in long-term (6 weeks) and short-term (12 days).
This indicated that the combined intervention T2DM mouse model was
stable. To shorten the modeling time and facilitate subsequent research,
the observation period of 12 days is feasible. Compared with the HFD +
STZ mouse model, the effect of HFD + DEX + STZ mouse model was
better, the success rate was higher and mortality was lower. Combined
intervention can effectively reduce insulin sensitivity and cause insulin
resistance in mice, and induce a stable hyperglycemia model.
5. Conclusion
A new mouse model of T2DM with insulin resistance and β-cell
damage could be established by combining intervention with a high-fat
diet, DEX and STZ. The optimized design can serve as a stable rodent
model for further research on the mechanisms and therapy of T2DM.
Funding
This work was supported by the Natural Science Foundation of China
(No. 81673248).
Ethical approval
All procedures performed in studies involving animal experiments
were comply with the ARRIVE guidelines and associated guidelines.
Compliance with ethical guidelines
Sha Liu, Lingling Ma, Xiaoyu Ren, Wenling Zhang, Dian Shi,Yanbei
Huo, Yupei Ba,Yana Bai and Ning Cheng declare that they have no
financial conflicts of interest. This manuscript is an original research
articles on basic medical science, all institutional and national guide­
lines for the care and use of laboratory animals were followed.
CRediT authorship contribution statement
Yana Bai and Ning Cheng: Conceptualization, Methodology;
Sha Liu: Methodology, Validation, Formal analysis Investigation,
Writing - Review & editing;
Lingling Ma: Validation, Formal analysis Investigation;
Xiaoyu Ren, Wenling Zhang, Dian Shi, Yanbei Huo and Yupei
Ba: Validation, Investigation.
6
Life Sciences 286 (2021) 120062
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