Hindawi Publishing Corporation

BioMed Research International

Volume 2014, Article ID 160980, 11 pages
http://dx.doi.org/10.1155/2014/160980

Research Article
Studies on the Antidiabetic Activities of
Cordyceps militaris Extract in Diet-Streptozotocin-Induced
Diabetic Sprague-Dawley Rats

Yuan Dong,1 Tianjiao Jing,1 Qingfan Meng,1 Chungang Liu,1 Shuang Hu,1
Yihang Ma,2 Yan Liu,1 Jiahui Lu,1 Yingkun Cheng,1 Di Wang,1 and Lirong Teng1,3
1
College of Life Science, Jilin University, Changchun 130012, China
2
College of Clinical Medicine, Jilin University, Changchun 130012, China
3
College of Life Science, Zhuhai College of Jilin University, Zhuhai 519000, China

Correspondence should be addressed to Di Wang; jluwangdi@gmail.com and Lirong Teng; tenglr@jlu.edu.cn

Received 3 December 2013; Revised 29 January 2014; Accepted 30 January 2014; Published 11 March 2014

Academic Editor: Gabriel F. Anhe

Copyright © 2014 Yuan Dong et al. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Due to substantial morbidity and high complications, diabetes mellitus is considered as the third “killer” in the world. A
search for alternative antidiabetic drugs from herbs or fungi is highly demanded. Our present study aims to investigate the
antidiabetic activities of Cordyceps militaris on diet-streptozotocin-induced type 2 diabetes mellitus in rats. Diabetic rats were orally
administered with water extract or alcohol extract at 0.05 g/kg and 2 g/kg for 3 weeks, and then, the factors levels related to blood
glucose, lipid, free radicals, and even nephropathy were determined. Pathological alterations on liver and kidney were examined.
Data showed that, similar to metformin, Cordyceps militaris extracts displayed a significant reduction in blood glucose levels
by promoting glucose metabolism and strongly suppressed total cholesterol and triglycerides concentration in serum. Cordyceps
militaris extracts exhibit antioxidative effects indicated by normalized superoxide dismutase and glutathione peroxidase levels. The
inhibitory effects on blood urea nitrogen, creatinine, uric acid, and protein revealed the protection of Cordyceps militaris extracts
against diabetic nephropathy, which was confirmed by pathological morphology reversion. Collectively, Cordyceps militaris extract,
a safe pharmaceutical agent, presents excellent antidiabetic and antinephropathic activities and thus has great potential as a new
source for diabetes treatment.

1. Introduction normalize blood glucose and fat levels, possess hypotensive

activity, and improve microcirculation [4]. Traditional ther-
Diabetes mellitus (DM) is characterized by chronic hypergly- apy only focuses on pancreatic islet function recovery and
caemia which is resulted by the defects of insulin secretion blood glucose regulation, which fails to control the diabetic
or action. Diabetes patients suffer with a series of metabolic complications [5]. As reported previously, insulin injection
disorders in carbohydrate, fat, and proteins [1]. Noninsulin- and some oral antihyperglycemic agents, such as metformin
dependent diabetes mellitus (NIDDM), caused by insulin and pioglitazone, display undesirable adverse effects [6].
resistance, is known as the most common form of dia- Pioglitazone induces hepatocellular-cholestatic liver injury
betes (type 2 diabetes) [2]. According to statistics, till 2025, and metformin causes diarrhea and nausea or vomiting [7].
8 billion people in the whole world will suffer with type Additionally, weight gain, hypoglycemia, edema, gastroin-
2 diabetes. Additionally, various complications including testinal disturbances, and insulin resistance are observed in
cardiovascular disease, nephropathy, neuropathy, retinopa- diabetes patients who receive long-term insulin treatment [8].
thy, and hyperlipemia are observed in most diabetes patients Meanwhile, diabetes mellitus requires lifelong medication,
[3]. and the economic burden of patients should receive attention
As a cosmopolitism tough problem, no satisfactory ther- [9]. Due to the limitation of existing antidiabetic agents, a
apeutic regimen can cure diabetes although most of them search for alternative treatment is highly demanded.
2 BioMed Research International

Herbal medicine turns out to be a valuable reservoir for content of the total polysaccharides was 163 ± 2.32 mg/g in
novel drugs due to its few side effects [10]. Amount of research WE and 104 ± 1.27 mg/g in AE.
demonstrated that natural products possess antidiabetic
activity with less adverse effects and show great auxiliary ther- 2.3. In Vivo Experiment in Animal Model of Diabetes. Exper-
apeutic effect on complications [11, 12]. Cordyceps militaris, an imental protocol was approved by the Lab Animal Centre
anamorph of Cordyceps sinensis, is advertised as a Chinese of Jilin University (licence number SCXK-(JI) 2006-0001).
herb with antioxidant [13], immunomodulatory [14], anti- Sprague-Dawley male rats weighing 120–140 g were housed
cancer, and anti-inflammatory pharmacological [15] effects. in groups of two in clear plastic cages and maintained on a
Cordyceps polysaccharides, the richest and most important 12 h light/dark cycle (lights on 07:00–19:00 h) at 23 ± 1∘ C with
activity component, display a hypoglycemic activity [16]. water and food available ad libitum.
Additionally, several studies have shown that water extracts The experimental protocol for diabetic rat model estab-
of Cordyceps militaris possess notable activity via increment lishment and drug administration was shown in Figure 1.
of insulin secretion and cholinergic activation in normal To produce experimental model of diabetes, 42 male Sprague-
Wistar rats [17, 18]. Excitingly, separated research finds that Dawley rats were administrated with a modified high fat
Cordyceps militaris can be used for kidney protection [19]. high sucrose diet (HFHSD; 68.8% standard chow, 20%
However, the regulatory effects of polysaccharide-enriched sucrose, 10% lard, 0.2% cholesterol, and 1% salt mixture;
fraction of Cordyceps militaris on Sprague-Dawley rats with purchased from the Lab Animal Centre of Jilin University,
diabetes have not been reported yet. Jilin, China) [22] for 8 weeks followed by the injection of
We therefore hypothesized that Cordyceps militaris 30 mg/kg streptozotocin (STZ) for 3 days (i.p., once per day)
extracts may show antidiabetic, hypolipidemic, and even [23]. During the experiment, 5% glucose solution was fed
antinephritic effects. To test this hypothesis, the present study to rats 4 h after STZ injection to prevent hypoglycaemia.
aims to investigate the related biological activities of Cordy- Rats with fasting serum glucose levels between 11 mmol/L
ceps militaris extracts via in vivo experiments. After treat- and 26 mmol/L were identified as severe diabetic groups
ment with polysaccharide-enriched fractions of Cordyceps for further study [24]. Another 7 male Sprague-Dawley rats
militaris, the changes of serum fasting glucose levels, pyruvate feeding with normal diet for 8 weeks and injected with citrate
kinase activity, triglyceride (TG), and total cholesterol in buffer for 3 days served as control group (CT) which were
experimental diabetic Sprague-Dawley rats were detected. treated with normal saline orally for another 3 weeks. All
Several indexes associated with oxidation resistance and diet-STZ-induced diabetic rats were separated for 6 groups
hypolipidemic activity were also determined. Furthermore, randomly as follows and received drug administration for 3
the therapeutic effects of Cordyceps militaris extracts on weeks (once a day):
diabetic nephropathy were detected through histopathologic
morphology observation and four indexes analysis including diabetic model group (DM; 𝑛 = 7): treatment with
blood urea nitrogen (BUN), uric acid (UA), creatinine, and normal saline orally;
urine protein.
metformin (DH) group (𝑛 = 7): treatment with
120 mg/kg metformin orally;
2. Methods
low dose AE treated group (𝑛 = 7): treatment with
2.1. Submerged Fermentation of Cordyceps militaris. Cordy- 0.05 g/kg AE orally;
ceps militaris (NBRC9787; obtained from National Biological high dose AE treated group (𝑛 = 7): treatment with
Resource Center, Japan) was cultured in a rotary shaker 2 g/kg AE orally;
incubator (10 L, Biostat B; Germany) at 150 rpm for 5 days and
the cultured temperature was 26∘ C. The cultured medium was low dose WE treated group (𝑛 = 7): treatment with
as follows: glucose, 20 g/L; peptone, 10 g/L; yeast extract pow- 0.05 g/kg WE orally;
der, 18 g/L; KH2 PO4 , 3 g/L; MgSO4 ⋅7H2 O, 3 g/L; (NH4 )2 SO4 ,
high dose WE treated group (𝑛 = 7): treatment with
10 g/L; ZnCl2 , 0.01 g/L; Vitamin B1 , 0.24 g/L. The mycelium
2 g/kg WE orally.
pellets were harvested and lyophilized for further using.
After 3-week treatment, food intake, water intake, and
2.2. Cordyceps militaris Extract Preparation. As reported urine excretion in all rats were monitored within 16 h. Blood
previously [20], the water and the alcohol extract from and urine samples were collected, and the fasting serum
Cordyceps militaris were prepared as follows: 100 g mycelial glucose, pyruvate kinase (PK), superoxide dismutase (SOD),
powder was extracted two times in double distilled water at glutathione peroxidase (GSH-Px), triglycerides (TG), total
80∘ C for 3 h. After centrifuging at 5000 rpm for 10 min, using cholesterol, blood urea nitrogen (BUN), uric acid (UA),
Sevag reagent [V (n-butanol) : V (chloroform) = 1 : 4, 50 mL], creatinine, and urine protein levels were determined. All the
the proteins that existed in the extracts were removed [21]. assay kits were obtained from Nanjing Biotechnology Co. Ltd.
After concentration, the water extract (WE) was freeze-dried (Nanjing, China). After oral glucose tolerance test, animals
and stored in vacuum environment. Similarly, the alcohol were sacrificed by administration of 200 mg/kg pentobarbital;
extract (AE) was prepared using alcohol distillation at 60∘ C meanwhile, liver and kidney were collected and fixed in 4%
for 3 h followed by proteins removing and freeze-drying. The paraformaldehyde.
BioMed Research International 3

HFHSD feeding STZ injection Drug administration Test day OGTT test Sacrifice
(once a day) (once a day) Blood
HFHSD: 68.8% standard collection;
chow, 20% sucrose, Met. (120 mg/kg) Last food, water 2 g/kg Organ collection
10% lard, 0.2% 30 (mg/kg) AE (0.05 and 2 g/kg) treatment intake; glucose histopathological
cholesterol, and 1% salt treatment assessment
mixture WE (0.05 and 2 g/kg) urine
excretion;

8 weeks 3 days 3 weeks Monitoring 16 h 12 h fasting

within 120 min test

Figure 1: The experimental protocol for diet-STZ-induced diabetic rat model establishment and drug administration. Details are described
in Section 2.

2.4. Oral Glucose Tolerance Test (OGTT). As shown in 450

Figure 1, after 3-week treatment in diet-STZ-induced diabetic
rats, an oral glucose tolerance test (OGTT) was performed. 400
After a 12 h fast, all the experimental rats were received
physiological saline, metformin, AE, or WE, respectively, as 350
described above; 30 min later, 2 g/kg of glucose was orally

Bodyweight (g)
given to all the rats. Blood samples were collected at 0, 30, 60, 300
and 120 min to detect the blood glucose levels using Glucose
Assay Kit (Nanjing Biotechnology Co. Ltd., Jiangsu, China). 250 ##
Calculation of the area under the blood glucose curve (AUC)
was made according to (I) [25]: 200 ∗

AUC = (basal glycaemia + glycaemia 0.5 h) × 0.25 150

+ (glycaemia 0.5 h + glycaemia 1 h) × 0.25 (I)

100
+ (glycaemia 1 h + glycaemia 2 h) × 0.5. HFHSD feeding 3-day Drug administration
8 weeks STZ 3 weeks
injection
2.5. Histopathological Examination. Collected tissues were
CT DH
immerged in 4% paraformaldehyde for 48 h and then dehy-
AE (0.05 g/kg) AE (2 g/kg)
drated in gradient ethanol (50%, 70%, 80%, 90%, 95%, and WE (2 g/kg)
WE (0.05 g/kg)
100%) step by step. Samples were immerged in xylene for DM
30 min and incubated with first paraffin at 65∘ C overnight.
After embedding in wax, tissues were cut into serial sections Figure 2: Bodyweight changes in all separated experimental groups
at 5 𝜇m thickness using microtome (Leica, Germany) and during the whole process. Data are expressed as mean ± SD (𝑛 = 7)
spread over microscopy slides. Sections were deparaffinized and analyzed using one-way ANOVA followed by Dunn’s test. ## 𝑃 <
with fresh xylene for 10 min, hydrated with gradient ethanol 0.01 versus controls. ∗ 𝑃 < 0.05 versus DM group. CT: control rats;
(100%, 90%, 80%, and 70%), and then washed with double DM: diabetic rats; DH: metformin; AE: alcohol extract; WE: water
extract.
distilled water for three times. The sections were analyzed
via haematoxylin and eosin staining (H&E staining) [26]
and examined by a light microscope digital camera (Nikon
Instruments, Tokyo, Japan). excretion and water intakes in diet-STZ-induced diabetic rats
compared with DM rats; however, no significant changes in
2.6. Statistical Analysis. All values were expressed as mean food intakes were observed (Table 1; 𝑃 > 0.05). Compared
± SD. One-way analysis of variance (ANOVA) was used to with CT, the growth of diet-STZ-induced diabetic rats was
detect statistical significance followed by post hoc multiple inhibited strongly (𝑃 < 0.01; Figure 2); however, after 3-
comparisons (Dunn’s test). A value of 𝑃 < 0.05 was consid- week 2 g/kg AE and WE and 120 mg/kg DH treatment, the
ered to be significant. bodyweight was increased significantly compared with DM
group (𝑃 < 0.05; Figure 2, Table 1).
3. Results
3.2. Hypoglycemic Effects of Cordyceps militaris Extracts in
3.1. Bodyweight, Food and Water Intakes, and the Urine Excre- Diet-STZ-Induced Diabetic Rats. To evaluate the hypoglyce-
tion Monitoring. Compared with CT group, DM rats con- mic effects of Cordyceps militaris extracts, the changes in
sumed more food and water (𝑃 < 0.05; Table 1); meanwhile, fasting blood glucose (FBG) levels and PK activity were mea-
more urine excretion in DM rats was noted (𝑃 < 0.05; sured. Fasting blood glucose concentration in DM rats was
Table 1). Similar to DH-administrated rats, both AE and WE 11.1 mmol/L higher than that of CT; while, 120 mg/kg DH and
treatment at 0.05 g/kg and 2 g/kg strikingly decreased urine 0.05 g/kg AE and WE treatment resulted in a 46.1%, 86.3%,
4 BioMed Research International

Table 1: Results on the bodyweight gain, food intake, water intake, and urine excretion in each experimental group.

Group Dose (g/kg/d) Bodyweight gain/(g) Water intake/(mL) Food intake/(g) Urine excretion/(mL)
CT — 5.58 ± 0.27 55.00 ± 10.48 9.17 ± 1.60 36.67 ± 13.70
DM — 6.61 ± 1.29 102.00 ± 5.41b 16.25 ± 0.50b 82.50 ± 5.00b
DH treated 0.12 37.06 ± 2.38a 37.50 ± 9.58a 4.75 ± 1.50a 22.50 ± 2.65a
0.05 6.75 ± 0.68 54.17 ± 32.31a 6.33 ± 2.50 29.60 ± 3.55a
AE treated
2 25.82 ± 2.14a 50.00 ± 17.03a 5.41 ± 1.02 37.60 ± 19.03a
0.05 38.49 ± 3.86a 46.67 ± 5.77a 7.67 ± 2.51 21.33 ± 6.11a
WE treated
2 13.34 ± 1.57 53.33 ± 15.27a 5.67 ± 0.57 56.60 ± 15.27a
Data are expressed as mean ± SD (𝑛 = 7/group) and analyzed using ANOVA followed by Dunn’s test.
a
Statistical significance compared with DM (𝑃 < 0.05); b statistical significance compared with CT (𝑃 < 0.05).
CT: control rats; DM: diabetic rats; DH: metformin; AE: alcohol extract; WE: water extract.

25 400
## ∗∗∗
Fasting blood glucose (FBG) (mmol/L)

20 320

Pyruvate kinase activity (U/L) ∗∗

15 240
∗

10 ∗∗ ∗∗ 160

∗∗ ∗∗ ##
5 80
∗∗

0 0
CT DM DH 0.05 2 0.05 2 CT DM DH 0.05 2 0.05 2
Met. (120 mg/kg) AE (g/kg) WE (g/kg) Met. (120 mg/kg) AE (g/kg) WE (g/kg)
HFHSD + STZ (30 mg/kg) HFHSD + STZ (30 mg/kg)
(a) (b)

Figure 3: Diabetic rat model was established by 8-week HFHSD administration followed by thrice intraperitoneal injection of low doses of
30 mg/kg STZ. Nontreated rats served as control group. Diet-STZ-induced diabetic rats were treated with or without 120 mg/kg metformin
(DH) and Cordyceps militaris extracts at various doses for another 3 weeks. The changes fasting plasma glucose level (a) and pyruvate kinase
activity (b) were determined. Data are expressed as mean ± SD (𝑛 = 7) and analyzed using one-way ANOVA followed by Dunn’s test.
## ∗ ∗∗ ∗∗∗
𝑃 < 0.01 versus controls, 𝑃 < 0.05, 𝑃 < 0.01, and 𝑃 < 0.001 versus DM group. CT: control rats; DM: diabetic rats; AE: alcohol
extract; WE: water extract.

and 85.2% reduction, respectively (𝑃 < 0.01; Figure 3(a)). AE The calculated AUC values for glucose response during
and WE administration markedly enhanced the lower PK the OGTT revealed a striking increment in DM group
activity in diet-STZ-induced diabetic rats (𝑃 < 0.05; (25.69 ± 0.46 mmol L−1 h) compared with CT group (6.58 ±
Figure 3(b)). Especially, 2 g/kg AE treatment increased nearly 0.31 mmol L−1 h) (𝑃 < 0.01; Figure 4(b)). However, com-
4-fold PK activity compared with DM rats (𝑃 < 0.001; pared with DM group, AE and WE treatment at 0.05 g/kg
Figure 3(b)). and 2 g/kg showed a significant reduction in AUC (𝑃 < 0.01;
3.3. Oral Glucose Tolerance Test. In order to avoid false Figure 4(b)).
positive obtained from FBG, OGTT was performed as second
diagnostic indices [27]. Compared with CT rats, dramatically 3.4. Hypolipidemic Effects of Cordyceps militaris Extracts in
higher fasting blood glucose concentration was noted in diet- Diet-STZ-Induced Diabetic Rats. Hyperlipidemia is a com-
STZ-induced diabetic rats from 0 min up to 120 min (𝑃 < mon accompanied disease related to diabetes [28]. Data
0.05; Figure 4(a)) indicating an impaired glucose tolerance showed that 0.05 g/kg WE strikingly reduced serum TG
(IGT) state. AE and WE significantly prevented the blood concentration to 72.43% and total cholesterol concentration
glucose levels shooting up, especially at 60 min and 120 min to 55.02% compared with DM rats (𝑃 < 0.01; Figure 5). AE
(𝑃 < 0.05; Figure 4(a)). treatment suppressed high total cholesterol concentration up
BioMed Research International 5

20

##
16 30
Blood glucose levels (mmol/L)

##
## ## 25
12 ∗∗
∗∗
##
∗∗ ∗∗ 20

AUC (mmol L−1 h)

∗∗
8 ∗∗ ∗∗
∗∗
∗∗ ∗∗ ∗∗
15
∗∗ ∗∗ ∗∗ ∗∗
∗∗ ∗∗ ∗∗
4
∗ ∗∗
∗∗
∗∗ 10
∗∗

0
0.0 0.5 1.0 1.5 2.0 5
Time (h)

CT DH 0
CT DM DH 0.05 2 0.05 2
AE (0.05 g/kg) AE (2 g/kg)
Met. (120 mg/kg) AE (g/kg) WE (g/kg)
WE (0.05 g/kg) WE (2 g/kg)
DM HFHSD + STZ (30 mg/kg)
(a) (b)

Figure 4: Blood glucose level (a) and AUC of OGTT (2 g glucose kg−1 BW) (b) in normal and diabetic rats in OGTT experiment. Data are
expressed as mean ± SD (𝑛 = 7) and analyzed using one-way ANOVA followed by Dunn’s test. ## 𝑃 < 0.01 versus controls, ∗∗ 𝑃 < 0.01 versus
DM group. CT: control rats; DM: diabetic rats; AE: alcohol extract; WE: water extract.

4.2 4.2
###
##
3.5 3.5
Total cholesterol (mmol/L)

∗
∗
2.8 2.8
TG (mmol/L)

∗∗∗
∗∗
∗∗ ∗∗∗ ∗∗∗
2.1 2.1

1.4 ∗∗∗ 1.4

0.7 0.7

0.0 0.0
CT DM DH 0.05 2 0.05 2 CT DM DH 0.05 2 0.05 2
Met. (120 mg/kg) AE (g/kg) WE (g/kg) Met. (120 mg/kg) AE (g/kg) WE (g/kg)
HFHSD + STZ (30 mg/kg) HFHSD + STZ (30 mg/kg)
(a) (b)

Figure 5: Both AE and WE treatment significantly reduce the levels of serum TG (a) and total cholesterol (b) in diet-STZ-induced diabetic
rats. Data are expressed as mean ± SD (𝑛 = 7) and analyzed using one-way ANOVA. ## 𝑃 < 0.01 and ### 𝑃 < 0.001 versus controls, ∗∗ 𝑃 < 0.01
and ∗∗∗ 𝑃 < 0.001 versus DM group. CT: control rats; DM: diabetic rats; AE: alcohol extract; WE: water extract; TG: triglycerides.
6 BioMed Research International

300 600
∗∗
∗∗ ∗∗
250 500

###
SOD activity (U/mL)

200 400

GSH-Px (𝜇mol/L)
∗∗

150 300 ∗∗

100 200

50 100

0 0
CT DM DH 0.05 2 0.05 2 CT DM DH 0.05 2 0.05 2

Met. (120 mg/kg) AE (g/kg) WE (g/kg) Met. (120 mg/kg) AE (g/kg) WE (g/kg)
HFHSD + STZ (30 mg/kg) HFHSD + STZ (30 mg/kg)
(a) (b)
Figure 6: Diet-STZ-induced diabetic rats were treated with or without 120 mg/kg metformin (DH) and Cordyceps militaris extracts at various
doses for 3 weeks. The serum SOD activity (a) and serum GSH-Px level (b) were detected. Data are expressed as mean ± SD (𝑛 = 7) and
analyzed using one-way ANOVA followed by Dunn’s test. ### 𝑃 < 0.001 versus controls, ∗ 𝑃 < 0.05 and ∗∗ 𝑃 < 0.01 versus DM group. CT:
control rats; DM: diabetic rats; AE: alcohol extract; WE: water extract; SOD: superoxide dismutase; GSH-Px: glutathione peroxidase.

to 53.42%; however, no inhibitory effect on TG concentration groups were observed in liver tissues indicating Cordyceps
was noted (𝑃 < 0.01; Figure 5). militaris is safe for animal treating (Figure 8). Moreover, data
revealed that STZ caused severe injury in kidney. A large
3.5. Antioxidative Effects of Cordyceps militaris Extracts in amount of inflammatory cell infiltration in renal interstitium
Diet-STZ-Induced Diabetic Rats. Oxidative stress is consid- and the atrophy of renal tubule cells was noted in DM group,
ered as a major pathogenesis in diabetes-related complica- whereas pathological morphology was reversed after 3-week
tions [29]. Serum SOD and GSH-Px levels were detected; DH, AE, and WE treatment (Figure 8).
among them, only abnormal SOD activity was noted in DM
rats (𝑃 < 0.05; Figure 6(a)). AE and WE treatment resulted 4. Discussion
in an increment of SOD activity and GSH-Px level compared
with DM rats (𝑃 < 0.05; Figure 6(b)). Meanwhile, DH only Diabetes has become the third “killer” in the world following
increased serum GSH-Px level in diabetic rats (𝑃 < 0.05; cancer, cardiovascular and cerebrovascular diseases. Due to
Figure 6(b)). various pathologic changes, amount of diabetic complica-
tions occur in blood vessels, cranial and peripheral nerves,
3.6. Antidiabetic Nephropathic Effects of Cordyceps militaris and skin [31]. Retinopathy and nephropathy are also consid-
Extracts in Diet-STZ-Induced Diabetic Rats. The levels of ered as high frequency complications caused by the abnormal
serum BUN, UA, creatinine, and urine protein, considered thickening of the basement membrane in capillaries [32]. The
as sensitive indexes for kidney injury [30], were enhanced antihyperglycemic and antidyslipidemic effects of Cordyceps
significantly in diet-STZ-induced diabetic rats (𝑃 < 0.05; militaris water extracts have already been studied in db/db
Figure 7). Both AE and WE treatment reduced nearly twofold mice [33]. As reported previously, the polished rice cultivated
of serum BUN and protein concentration compared with DM with Cordyceps militaris reduces blood glucose levels and
rats (𝑃 < 0.05; Figures 7(a) and 7(c)). Additionally, only shows antioxidant effects in STZ-induced diabetic rats [27].
WE treatment suppressed the high UA and creatinine levels In our present study, the hypoglycemic effects related to fast-
in serum (𝑃 < 0.05; Figures 7(b) and 7(d)). Interestingly, ing blood glucose levels of Cordyceps militaris extracts were
the antinephropathic effect of Cordyceps militaris extracts is confirmed. Furthermore, the hypolipidemic, antioxidative,
much better than DH which only suppressed serum BUN and and antidiabetic nephropathic effects of Cordyceps militaris
creatinine levels in diet-STZ-induced diabetic rats (𝑃 < 0.05; extracts were observed in diet-STZ-induced diabetic rats.
Figures 7(a) and 7(d)). Through histopathological analysis in liver tissue, Cordyceps
militaris was confirmed as a pharmacological safe agent.
3.7. Histopathological Analysis. Histopathological sections In our present study, the results showed that both WE and
were performed to examine the in vivo toxicity of Cordyceps AE possess hypoglycemic effect similar to that of DH, and
militaris extracts and further confirm their antinephro- this effect may be partially related to the increment of glucose
pathic effects. No significant differences among experimental absorption indicated by the enhancement of PK activity [34].
BioMed Research International 7

6 40

#
5 ##
32

∗
4
BUN (mmol/L)

24 ∗∗

UA (mg/L)
∗
3
∗ 16
∗∗∗
2 ∗∗∗

8
1

0 0
CT DM DH 0.05 2 0.05 2 CT DM DH 0.05 2 0.05 2

Met. (120 mg/kg) AE (g/kg) WE (g/kg) Met (120 mg/kg) AE (g/kg) WE (g/kg)
HFHSD + STZ (30 mg/kg) HFHSD + STZ (30 mg/kg)
(a) (b)
2.5
125
#
2.0 ##
100
Protein concentration (g/L)

Creatnine (𝜇mol/L)

1.5 ∗∗ 75 ∗∗
∗∗
∗∗ ∗∗
1.0 50
∗∗
∗∗∗
0.5 25

0.0 0
CT DM DH 0.05 2 0.05 2 CT DM DH 0.05 2 0.05 2

Met. (120 mg/kg) AE (g/kg) WE (g/kg) Met. (120 mg/kg) AE (g/kg) WE (g/kg)
HFHSD + STZ (30 mg/kg) HFHSD + STZ (30 mg/kg)
(c) (d)

Figure 7: Diet-STZ-induced diabetic rats were treated with or without 120 mg/kg metformin (DH) and Cordyceps militaris extracts at various
doses for 3 weeks. Serum BUN (a), UA (b), protein concentration (c), and creatinine (d) levels in all groups were detected. Data are expressed
as mean ± SD (𝑛 = 7) and analyzed using one-way ANOVA followed by Dunn’s test. ## 𝑃 < 0.01 versus controls, ∗∗ 𝑃 < 0.01 versus DM group.
CT: control rats; DM: diabetic rats; DH: metformin; AE: alcohol extract; WE: water extract; BUN: blood urea nitrogen; UA: uric acid.

PK is the rate-limiting enzyme of glycolytic pathway which of glycogen synthase in 90% pancreatectomized rats [38].
can promote the metabolism of sugar [35]. Due to the disor- Further study will be performed to confirm this result.
der of lipid metabolism [36], serum concentrations of total Increased glucose levels result in glucose autooxidation
cholesterol and TG were enhanced significantly in DM rats; and autooxidative glycosylation of proteins [27]. Due to the
in contrast, both WE and AE normalized plasma lipid and unbalanced producing and scavenging, free radicals were
lipoprotein profile. Cordyceps militaris extracts may prevent accumulated in diabetic patients, which played a role in
the accumulation of fatty acid in liver [37]. Since Cordyceps the pathogenesis of the long-term complications of human
militaris extracts displayed no stimulating effect on insulin diabetes [39]. A deficiency of the antioxidant activity of SOD
(data not shown), their antidiabetic effect may not be related and GSH-Px in diabetes mellitus is related to higher concen-
to insulin secretion. Previous study reports that Cordyceps tration of peroxide [40]. Our data demonstrated that both
militaris water extract decreases fasting serum glucose levels AE and WE normalized SOD activity and enhanced GSH-
by increasing glucose disposal rates and fraction velocity Px levels compared with DM group. Our finding is consistent
8 BioMed Research International

Kidney Liver

CT

DM

DH (Met.)
120 mg/kg

AE

0.05 g/kg
HFHSD + STZ (30 mg/kg)

AE
2 g/kg

WE
0.05 g/kg

WE
2 g/kg

Figure 8: Histopathological analysis in liver and kidney collected from all experimental rats through H&E staining (x400). Arrows show
regions of inflammatory cell infiltration in renal interstitium. CT: control rats; DM: diabetic rats; DH: metformin; AE: alcohol extract; WE:
water extract.

with other researches reporting that Cordyceps militaris kidney damage [43]. Cordyceps militaris extracts displayed a
extracts reduce reactive oxygen species (ROS) and nitrogen strong inhibitory effect on BUN, UA, creatinine, and urine
species (RNS) generation caused by high glucose [41]. protein levels compared with DM rats. Interestingly, mesan-
Furthermore, diabetic nephropathy is reported as a seri- gial expansion, mesangial hypercellularity, and a thickened
ous microvascular complication of diabetes [42]. BUN, UA, glomerular basement membrane, known as the morpholog-
urine protein, and creatinine are traditional indexes for ical indicators of nephropathy [44], were not obvious in
BioMed Research International 9

model group. However, a large amount of inflammatory cell Acknowledgment

infiltration in renal interstitium and the atrophy of renal
tubule cells was noted in DM rats, which were strongly This work was supported by the twelfth Five-Year National
reversed after DH, AE, and WE treatment. The pathological key Technology R&D Program: no. 2012BAI29B00.
lesions of renal tubule may also be related to STZ which
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10 BioMed Research International

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