Pharmaceutical Biology

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Antidiabetic activity of Helicteres angustifolia root

Xuansheng Hu, Delin Cheng & Zhenya Zhang

To cite this article: Xuansheng Hu, Delin Cheng & Zhenya Zhang (2016) Antidiabetic
activity of Helicteres�angustifolia root, Pharmaceutical Biology, 54:6, 938-944, DOI:
10.3109/13880209.2015.1077871

To link to this article: https://doi.org/10.3109/13880209.2015.1077871

Published online: 11 Feb 2016.

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PHARMACEUTICAL BIOLOGY, 2016
VOL. 54, NO. 6, 938–944
http://dx.doi.org/10.3109/13880209.2015.1077871

RESEARCH ARTICLE

Antidiabetic activity of Helicteres angustifolia root

Xuansheng Hua, Delin Chengb and Zhenya Zhanga
a
Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki, Japan; bTaisei Kogyo Co., Ltd, Tokyo, Japan

ABSTRACT KEYWORDS
Context The root of Helicteres angustifolia L. (Sterculiaceae) has been used as folk herbal drug to Antidiabetic activity;
treat cancer, bacterial infections, inflammatory, and flu in China. However, there is no report on its diabetes; glucose uptake
antidiabetic activity.
Objective This study evaluates the antidiabetic activity of ethanol extract from H. angustifolia root. HISTORY
Materials and methods The promoting effect of H. angustifolia root ethanol extract (25, 50, and Received 7 February 2015
Revised 5 July 2015
100 mg/mL) on glucose uptake was evaluated using HepG2 cell, differentiated C2C12 myotubes,
Accepted 27 July 2015
and differentiated 3T3-L1 adipocytes. The antidiabetic activity of the extract was assessed in vivo Published online 9 February
using STZ-induced diabetic rats by orally administration of the extract (200 and 400 mg/kg b.w.) 2016
once per day for 28 d. Blood glucose, TG, TC, TP, HDL-C, UA, BUN, AST, ALT, insulin, and HOMA-IR
were analyzed.
Results The results showed that the extract increased glucose uptake in C2C12 myotubes and 3T3-
L1 adipocytes with an IC50 value of 79.95 and 135.96 mg/mL, respectively. And about 12%, 19%, and
10% (p 5 0.05) in HepG2 cells when compared with the control at the concentration of 25, 50, and
100 mg/mL, respectively. After 28 days’ treatment with the extract, significant reduction was
observed in blood glucose, HOMA-IR, TC, TG, UA, BUN, AST, and ALT levels, while the levels of TP
and HDL cholesterol increased.
Discussion and conclusion These results suggest that H. angustifolia root ethanol extract possess
potent antidiabetic activity, which is the first report on antidiabetic activity of this plant.

Introduction treat cancer, bacterial infections, inflammatory and flu in

Diabetes mellitus is a chronic metabolic disease char-
acterized by high blood glucose level. It is estimated that
the number of diabetics will increase up to 439 million in
2030 (Shaw et al. 2010), and more than 90% of the
diabetics are type 2 diabetics (Attele et al. 2002). Type 2
diabetes is characterized by insulin resistance and a
progressive decline in b-cell function (Laakso 2001). It
has been reported that insulin resistance is an important
factor in the pathogenesis of type 2 diabetes (Meier &
Bonadonna 2013), which is characterized not only by
decreased responsiveness of the peripheral target tissues
(such as liver, adipose tissue, and skeletal muscle) to
insulin but also by remarkable decrease in glucose uptake
and utilization (Yang et al. 2014). Thus, increasing
glucose uptake of the above three key tissues is one of the
effective therapeutic approaches for treating diabetes.
Helicteres angustifolia L. (Sterculiaceae), namely Shan-
Zhi-Ma in Chinese, is one of the traditional medicinal
plants which are wildly distributed in South China and
Southeast Asia. Helicteres isora L. has shown significant
antidiabetic activity (Chakrabarti et al. 2002). Helicteres
angustifolia root has been used as a folk herbal drug to
China (Jiangsu New Medical College 1986). It has also
been reported to possess antihepatic fibrosis activity
(Huang et al. 2012) and antiviral activity (Huang et al.
2013). Different classes of phytochemical constituents
have been isolated from the root of H. angustifolia (Chen
et al. 1994 2006), e.g. triterpenoids (betulinic acid,
oleanolic acid, helicteric acid, and methyl heliceate),
flavonoids (kaempferol 3-O-b-D-glucopyranoside,
5,8-dihydroxy-7,40 -dimethoxyflavone, takakin 8-O-b-D-
glucuronide 600 -methyl ester and takakin 8-O-b-D-
glucuronide 200 -sodium sulfate), phenolic acids (rosmari-
nic acid), quinines (mansonone E, F, H, and H methyl
ester), lignans (lariciresinol, lirioresinol-B and (+)-pinor-
esinol), and cucurbitacins (cucurbitacin D and J).
Among these phytochemical constituents, betulinic acid
(De Melo et al. 2009), oleanolic acid (Gao et al. 2007),
and rosmarinic acid (Jayanthy & Subramanian 2014)
have been shown to exhibit antidiabetic activity. Up to
now, however, there is no report on the antidiabetic
activity of H. angustifolia root. Therefore, this study was
to investigate whether the root of H. angustifolia
possessed antidiabetic activity or not.

CONTACT Zhenya Zhang zhang.zhenya.fu@u.tsukuba.ac.jp, Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1, Tennodai,
Tsukuba, Ibaraki 305-8572, Japan
ß 2016 Taylor & Francis

This study evaluates the glucose uptake activity of H.
angustifolia root ethanol extract in HepG2 cell, differ-
entiated C2C12 myotubes and differentiated 3T3-L1
adipocytes in vitro, and antidiabetic activity in STZ-
induced diabetic rats. The total flavonoid content,
cytotoxicity, and acute oral toxicity of the extract were
also discussed.
Materials and methods
Chemicals, reagents, and culture medium
CMC-Na, STZ, glibenclamide, and glucose CII-Test were
purchased from Wako Pure Chemical (Osaka, Japan).
Insulin, 3-isobutylmethylxanthine (IBMX), dexametha-
sone (DEX), DMSO, and DMEM essential medium were
purchased from Sigma Aldrich, Inc. (St. Louis, MO).
Horse serum (HS) was from Cosmo-bio (Tokyo, Japan).
Fetal bovine serum (FBS) was from Giboco (Grand
Island, NY), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphe-
nyltetrazolium bromide (MTT) were purchased from
Life Technologies (Grand Island, NY). All other chem-
icals were of analytical grade.
Plant material
The roots of H. angustifolia were collected in the rural
area near the city of Vientiane, Laos in June, 2013. The
collected plant was identified by Dr. Ende Liu from
Kunming Institute of Botany, Chinese Academy of
Sciences, Kunming, China, of which a voucher specimen
(KUN-2014-141) was deposited at the herbarium of the
same institute.
The dried H. angustifolia root (500 g) was powdered
and extracted with 70% ethanol (5 L  3 times) at room
temperature (25 ± 2 C) for 3 d. Then the extract was
concentrated and lyophilized to get powdered product
(yielding 10.8%, w/w). The obtained 70% ethanol extract
from H. angustifolia root was stored at 20 C until
further use.
Cell culture and differentiation
Murine C2C12 myoblasts and human hepatocarcinoma
cell line HepG2 were purchased from RIKEN
Bioresource Center (Tsukuba, Japan). Mouse 3T3-L1
adipocyte was purchased from JCRB Cell Bank (Osaka,
Japan). All cells were cultured in Dulbecco’s modified
Eagle’s medium (DMEM) containing 10% fetal bovine
serum (FBS) and 1% penicillin/streptomycin at 37 C and
5% CO2 incubator. The medium was changed every
2–3 d.
PHARMACEUTICAL BIOLOGY 939
Differentiation of C2C12 myoblasts to myotubes was
carried out according to Kang et al. (2012) with slight
modifications. Briefly, cells were reseeded in 12-well
plates at a density of 2  104 cells/mL. When the cells
were over 80% confluent, the medium was replaced with
DMEM containing 2% horse serum and was changed
every 2 d. Cells were used for glucose uptake assay until
myotubes being observed under microscope.
Differentiation of 3T3-L1 cells to adipocytes was
performed according to Zhang et al. (2014) with some
modifications. In brief, cells were reseeded in 96-well
plates at a density of 5  103 cells/well. When the cells
were over 80% confluent, the medium was replaced with
DMEM containing 1 mM dexamethasone, 0.5 mM
3-isobutyl-1-methylxanthine, 10 mg/mL insulin for 3 d.
Then the cells were incubated in DMEM containing 10%
FBS, 1% penicillin–streptomycin, and 10 mg/mL insulin
for 2 d. Finally, the cells were incubated in DMEM
containing 10% FBS and 1% penicillin–streptomycin for
5–9 d. The cells were used for glucose uptake assay until
80–90% of them changed into lipid droplets under
microscope.
Cytotoxicity assay
The cytotoxic effect of H. angustifolia root extract on
HepG2, C2C12, and 3T3-L1 cells was determined by
using MTT assay (Lu et al. 2011). Briefly, HepG2,
C2C12, and 3T3-L1 cells were seeded at a density of
5  103 cells/well in 96-well plate and incubated for 24 h,
and then incubated with various concentrations of
H. angustifolia extract for another 24 h. After incubation,
10 mL MTT (5 mg/mL) was added into each well,
incubated for 4 h, then the culture medium was removed
before the addition of 100 mL DMSO. After shaking for
10 min, the optical density (OD) was measured at 570
nm with a microplate reader (Bio-Rad, Tokyo, Japan).
Glucose uptake assay
Glucose uptake in HepG2 cells was determined accord-
ing to Lv et al. (2014) with slight modifications. In brief,
HepG2 cells were seeded at a density of 1  104 cells/well
in 96-well plate with some wells left blank. After the cells
reached confluence, the medium was replaced by DMEM
containing 0.2% BSA. After 12 h incubation, the cells
were treated with DMEM containing 0.2% BSA with or
without the designated concentrations of test samples for
another 24 h. Then 10 mL of medium was removed from
each well (being placed into a new 96-well plate)
into which 200 mL of glucose oxidase reagent (Glucose
CII-Test, Wako, Japan) was added. The plates were
further incubated at 37 C for 15 min and the optical
940 X. HU ET AL.
density (OD) was measured at 490 nm with a microplate
reader (Bio-Rad, Tokyo, Japan). The amount of glucose
uptake was calculated by subtracting the glucose concen-
trations of the blank wells from the remaining glucose in
the cell plated wells. Insulin (10 mg/mL) was utilized as
positive while sterilized water as negative control.
Glucose uptake in differentiated C2C12 myotubes and
differentiated 3T3-L1 adipocytes was determined accord-
ing to the method used by Shimokawa et al. (2000) with
some modifications. Briefly, the differentiated C2C12
myotubes and differentiated 3T3-L1 adipocytes were first
added into 96-well plates. After incubation with DMEM
containing 0.2% BSA for 12 h, the cells were treated with
various concentrations of test samples for 24 h. The
amount of glucose uptake was calculated as that used for
HepG2 cells. Insulin (10 mg/mL) and sterilized water
were also utilized as positive and negative controls.
Animals
Sprague–Dawley rats, 6-week-old, were obtained from
Japan SLC, Inc. (Shizuoka, Japan) and maintained in
Laboratory Animal Resource Center, University of
Tsukuba, Japan. The rats were kept in cages with three
rats in each cage under controlled conditions of
temperature (23 ± 1 C), humidity (55 ± 5%), and
12/12-h light/dark cycle. The rats had free access to tap
water and food before the experiments. All the animal
experiments were based on the guideline of the main-
tenance and use of laboratory animals at the Laboratory
Animal Resource Center of University of Tsukuba and
were approved by the Animal Experiments Committee,
University of Tsukuba (Approval number 14-344).
Acute oral toxicity study
Acute oral toxicity assay of ethanol extract from
H. angustifolia root was performed according to the
OECD Guideline 423. Overnight fasted Sprague–Dawley
rats were randomly divided into two groups with three
rats in each group. The control group was given distilled
water, and the experimental group was given the extract
(5 g/kg). The rats were observed for 24 h. After 14 d, the
rats were dissected and their major organs were used for
determination.
Induction of experimental diabetic rats
Induction of experimental diabetic rats was performed
according to the literature (Masiello et al. 1998).
Briefly, after 1 week of preliminary breeding, the rats
were fasted overnight and then were injected intraper-
itoneally with streptozotocin (dissolved in 0.1 M cold
citrate buffer, pH 4.5) at a dose of 50 mg/kg. After 3 d,
the fasting blood glucose of rats was measured by one
touch select glucometer (Sanwa Kagaku Kenkyusho,
Nagoya, Japan). The rats with the fasting blood glucose
greater than 11 mmol/L were regarded as diabetic and
used for the following antidiabetic study.
Animal experimental protocol
The STZ-induced diabetic rats were randomly divided
into five groups with six in each group. The groups were
as follows:
Group I (NC): Normal control rats treated with 0.5%
CMC-Na solution (p.o.).
Group II (DC): Diabetic control rats treated with
0.5% CMC-Na solution (p.o.).
Group III (DE200): Diabetic rats treated with extract
(200 mg/kg b.w. p.o.).
Group IV (DE400): Diabetic rats treated with extract
(400 mg/kg b.w. p.o.).
Group V (DG): Diabetic rats treated with glibencla-
mide (1 mg/kg b.w. p.o.).
The ethanol extract from H. angustifolia root and an
antidiabetic drug, glibenclamide were suspended in 0.5%
CMC-Na solution, respectively, which were given by oral
route (p.o.) to the rats once per day for 28 d according to
the above experimental design.
Plasma collection and biochemical analysis
On day 28, all the rats were fasted overnight and
anesthetized with isoflurane, and then blood was collected
and centrifuged at 3000 rpm and 4 C for 15 min to obtain
serum. The serum sample was then measured with an
automated biochemical analyzer (DRI-CHEM 7000V,
Fujifilm, Tokyo, Japan) for the determination of some
biochemical parameters including blood glucose, total
glycerides (TG), total cholesterol (TC), total proteins
(TP), HDL cholesterol (HDL-C), uric acid (UA), urea
nitrogen (UN), aspartate transaminase (AST), and alanine
transaminase (ALT). Serum insulin levels were deter-
mined using a commercial kit (Mercodia, Uppsala,
Sweden) according to the instructions of the manufac-
turer. Insulin resistance was assessed by Homeosasis
Model Assessment of Insulin Resistance (HOMA-IR)
which was calculated from fasting glucose (mmol/
L  fasting insulin (mU/mL)/22.5 (Matthews et al. 1985).
Determination of total flavonoid content
The total flavonoid content was measured by using
aluminum chloride colorimetric assay (Yao et al. 2013).
 PHARMACEUTICAL BIOLOGY 941

HepG2 C2C12 3T3-L1

In brief, 1 mL extract or rutin at various concentration 140
was mixed with 4 mL 70% aqueous ethanol, and then 120
0.5 mL NaNO2 (5%, w/v) was added. After 6 min, 0.5 mL

Cell viability (%)

100
AlCl3 (10%, w/v) and 3 mL NaOH (1 M) were added, 80
followed by the addition of distilled water to reach
60
10 mL. After the solution was mixed and incubated for
40
15 min at 25 C, the absorbance was read at 510 nm using
20
a UV-1800 spectrophotometer (Shimadzu, Tokyo,
0
Japan). The total flavonoid content of in the ethanol 25 50 100 200 400
extract was calculated based on the standard curve Concentration (μg/mL)
obtained with rutin and expressed as rutin equivalent
(RE) in mg/g of dry sample. Figure 1. Cytotoxicity of ethanol extract of H. angustifolia root at
different concentrations on HepG2 cells, C2C12 cells, and 3T3-L1
cells.
Statistical analysis

All the data were expressed as mean ± SD, and ANOVA 250
HepG2 C2C12 3T3-L1

followed by Duncan’s multiple range test (DMRT) was

used for statistical analysis by using SPSS software 200 ∗∗

Glucose uptake (% of control )

∗∗ ∗
(version 17.0, SPSS Inc., Chicago, IL). Significance was ∗∗
assumed if p 5 0.05. 150 ∗
∗∗
∗ ∗ ∗

Results 100

Cytotoxicity assay
50
The cytotoxicity of the ethanol extract from
H. angustifolia root on HepG2, C2C12, and 3T3-L1 0

cells was assessed by MTT assay (Figure 1). At the Control 25 50 100 Insulin

concentration of 100 mg/mL, the cell viability of the Concentration (μg/mL)

extract was about 96% for HepG2, 114% for C2C12, and
103% for 3T3-L1 cells. Therefore, different doses of the
ethanol extract from H. angustifolia root, i.e. 25, 50, and
100 mg/mL were employed in the following glucose
uptake experiments.
Figure 2. Effects of ethanol extract of H. angustifolia root at
different concentrations on glucose uptake in HepG2 cells,
C2C12 myotubes, and 3T3-L1 adipocytes. Insulin and sterilized
water were utilized as positive and negative controls. *p50.05,
**p50.01 compared to negative control.

Glucose uptake assay
The effect of the ethanol extract on glucose uptake in
HepG2 cells, C2C12 myotubes, and 3T3-L1 adipocytes is
illustrated in Figure 2. The extract increased glucose
uptake in C2C12 myotubes and 3T3-L1 adipocytes with
an IC50 value of 79.95 and 135.96 mg/mL, respectively.
And about 12, 19, and 10% (p 5 0.05) in HepG2 cells
when compared with the control at the concentration of
25, 50, and 100 mg/mL, respectively. As a contrast,
insulin, as a positive control at 10 mg/mL, increased
glucose uptake in HepG2 cells, C2C12 myotubes, and
3T3-L1 adipocytes with a value of about 80, 91, and 75%,
respectively.
The above results indicate that the extract has
promoting effect on HepG2 cells, differentiated C2C12
myotubes and differentiated 3T3-L1 adipocytes which
represent the three key tissues involving in glucose
homeostasis, such as liver, muscle, and adipose tissue.
Acute oral toxicity study
During the acute oral toxicity experiments, oral admin-
istration of the extract at a dose of 5 g/kg did not produce
any signs of toxicity, and no death occurred. This
observation aroused us to seek more in depth facts
through the following long-term antidiabetic experiments.
Effect on blood glucose level, insulin level, and
HOMA-IR
As shown in Table 1, the diabetic rats were found to have
significantly higher fasting blood glucose concentrations,
insulin concentrations, and HOMA-IR than the normal
942 X. HU ET AL.

Table 1. Effect of ethanol extract of Helicteres angustifolia root on blood glucose, insulin, and HOMA-IR in the control and diabetic rats
(n ¼ 6) after 28 days’ experiments.
NC DC DE200 DE400 DG
a a a
Blood glucose (mmol/L) 10.91 ± 0.31 31.47 ± 0.30 24.83 ± 0.45 23.86 ± 0.25 22.15 ± 1.30a
Serum insulin (mU/mL) 43.54 ± 6.70 7.24 ± 0.38a 6.71 ± 0.16 6.98 ± 0.22 6.88 ± 0.22
HOMA-IRb 23.95 ± 1.30 10.13 ± 0.56a 7.41 ± 0.27a 7.41 ± 0.29a 6.15 ± 0.26a
Grouping of the animals: NC, normal control rats treated with 0.5% CMC-Na solution; DC, diabetic control rats treated with 0.5% CMC-Na solution; DE200,
diabetic rats treated with extract (200 mg/kg); DE400, diabetic rats treated with extract (400 mg/kg); DG, diabetic rats treated with glibenclamide (1 mg/kg).
a
p 5 0.01 compared with the NC group.
b
HOMA-IR, Homeosasis Model of Assessment-Insulin Resistance calculated from fasting glucose (mmol/L  fasting insulin (mU/mL)/22.5 (Matthews et al.,
1985).

Table 2. Effect of ethanol extract of H. angustifolia root on serum biochemical parameters in the control and diabetic rats (n ¼ 6) after
28 days’ experiments.
NC DC DE200 DE400 DG
a a a
ALT (mg/dL) 24.50 ± 2.26 83.67 ± 1.45 71.20 ± 2.2 64.67 ± 1.73 55.00 ± 2.73a
AST (mg/dL) 62.67 ± 2.55 164.33 ± 3.54a 145.80 ± 5.72a 112.83 ± 2.10a 103.00 ± 2.19a
TP (mg/dL) 6.40 ± 0.17 4.28 ± 0.12a 5.16 ± 0.07a 5.48 ± 0.11a 5.68 ± 0.70a
UA (mg/dL) 1.15 ± 0.08 4.27 ± 0.24a 2.62 ± 0.12a 2.10 ± 0.06a 1.65 ± 0.09
BUN (mg/dL) 17.47 ± 0.31 48.1 ± 1.50a 44.1 ± 0.56a 38.75 ± 1.14a 32.75 ± 1.04a
TC (mg/dL) 46.33 ± 1.93 234.83 ± 2.63a 132.60 ± 8.15a 124.67 ± 4.21a 86.83 ± 2.48a
TG (mg/dL) 77.67 ± 4.22 346.00 ± 7.38a 255.40 ± 4.41a 236.17 ± 4.50a 131.17 ± 7.85a
HDLC (mg/dL) 69.50 ± 1.57 32.33 ± 1.54a 40.20 ± 1.24a 47.33 ± 1.76a 59.50 ± 1.61a
ALT: alanine transaminase; AST: aspartate transaminase; BUN: blood urea nitrogen; HDL-C: HDL cholesterol; TC: total cholesterol; TG: total glycerides;
TP: total proteins; UA: uric acid.
Grouping of the animals: NC: normal control rats treated with 0.5% CMC-Na solution; DC: diabetic control rats treated with 0.5% CMC-Na solution; DE200:
diabetic rats treated with extract (200 mg/kg); DE400: diabetic rats treated with extract (400 mg/kg); DG: diabetic rats treated with glibenclamide (1 mg/kg).
a
p 5 0.01 compared with the NC group.

control rats. After 28 days’ administration of the extract
(200 and 400 mg/kg), the fasting blood glucose concen-
trations were observed to decrease when compared with
the diabetic rats (p 5 0.05). This observation suggests
that the extract has hypoglycemic effect on STZ-induced
diabetic rats, resulting in decrease in the insulin
resistance.
Effect on serum lipid profile, renal, and hepatic
parameters
As seen from Table 2, the TG, TC, UA, BUN, AST, and
ALT levels of the diabetic rats were higher while TP and
HDL-C were lower than those of the normal rats. After
28 days’ administration of the extract (200 and
400 mg/kg) and glibenclamide (1 mg/kg), however, the
TG, TC, UA, BUN, AST, and ALT levels were signifi-
cantly decreased; in contrast, significant increases in TP
and HDL-C levels were detected in the tested rats when
compared with diabetic rats (p 5 0.01).
Determination of total flavonoid content
The total flavonoid content in the ethanol extract of H.
angustifolia root was determined according to the
calibration curve equation (y ¼ 0.0012x – 0.0115,
R2 ¼ 0.9991) and expressed in rutin equivalents (RE).
Result showed that the extracts contained 430.70 ±
0.92 mg/g of total flavonoids.
Discussion
The results from this study indicate that the ethanol
extract of H. angustifolia root has promoting effect on
HepG2 cells, differentiated C2C12 myotubes, and
differentiated 3T3-L1 adipocytes that represent the
three key tissues involving in glucose homeostasis liver,
muscle, and adipose tissue, respectively. The extract at a
dose of 100 mg/mL could significantly promote the
glucose uptake in C2C12 myotubes (p50.01), HepG2
cells (p50.05), and differentiated 3T3-L1 adipocytes
(p50.05). This phenomenon is similar to the positive
control of insulin. This study for the first time confirms
that the ethanol extract of H. angustifolia root has
antidiabetic effects by stimulating glucose uptake in
HepG2 cells, C2C12 myotubes, and 3T3-L1 adipocytes.
Hypertriglyceridaemia and hypercholesterolaemia
have been reported to be the primary factors of diabetic
state involving in the development of atherosclerosis and
coronary heart disease which are the secondary compli-
cations of diabetes (Ananthan et al. 2003). Results from
this study indicate that the extract administration could
significantly reduce TG and TC levels while significantly
increase HDL-C level in the STZ-induced diabetic rats.

Therefore, it can be inferred that the extract has a
potential for improving serum lipid abnormalities in
diabetic conditions and thus decreasing the risk of
atherosclerosis and coronary heart disease.
Liver is the center of glucose metabolism, which not
only maintains normal blood glucose level but also
provides glucose as energy to organs. ALT and AST are
enzymatic markers of liver function and they are usually
used to signal liver damage in clinics. Compared with the
normal rats, the diabetic rats had significantly higher
levels of ALT and AST while lower level of TP (Table 2).
The results showed that the diabetic rats were in a liver
injury state. After administration of the extract, the ALT
and AST levels were found to be significantly decreased
in the STZ-induced diabetic rats while TP was signifi-
cantly increased, implying that the extract could poten-
tially protect liver against diabetic damage.
Kidney is an important organ to remove the metabolic
wastes such as blood urea nitrogen (BUN) and creatinine
from body, thereby helping to maintain body homeo-
stasis of above mentioned substances (Ramachandran
et al. 2012). The results from Table 2 show that the
extract administration could significantly decrease BUN,
an indicator of renal function. It is well known that
diabetic renal disease is one of the diabetic complica-
tions. Therefore, results from this study imply that the
ethanol extract has potential for treating diabetic
complications.
UA is the last metabolite of the purine catabolic
pathway in humans. High UA content in blood is a
characteristic of hyperuricemia (Haidari et al. 2008).
Reducing UA level is one of the effective ways for
preventing and treating gout. The results (Table 2) show
that administration of the extract could significantly
decrease UA, implying that the extract has a potential for
treating gout.
HOMA-IR is a reliable and useful parameter for
assessing insulin resistance in patients with type 2
diabetes (Okita et al. 2013). Results from this study
demonstrate that administration of the extract did not
lead to increased insulin levels, but could significantly
lower the blood glucose level and HOMA-IR. These
results are in consistence with the results from glucose
uptake assay, proving that the extract could increase
glucose uptake in insulin-target tissues and improve
insulin resistance. So it can be concluded that the
mechanism involving in the promoting effect of the
extract on antidiabetic activity is probably realized by
reducing insulin resistance and also by increasing
glucose uptake in insulin-target tissues.
The results from acute oral toxicity experiments
indicate that the ethanol extract of H. angustifolia root
is non-toxic even at a high oral dose of 5 g/kg. In the
PHARMACEUTICAL BIOLOGY 943
long-term antidiabetic test, doses of 200 and 400 mg/kg
were applied in the experiments.
From the results of animal experiments, the extract
exhibited antidiabetic effect on the STZ-induced diabetic
rats and could decrease the insulin resistance. In this
study, no significant change in body weight was observed
between the treatment group and the control group (data
not shown). This is also the first study to disclose the
antidiabetic activity of the ethanol extract from H.
angustifolia root in STZ-induced diabetic rats.
It is commonly recognized that certain flavonoids and
triterpenoids from plant exhibit antidiabetic activity
(Perez et al. 1998). In previous studies, several flavonoids
and triterpenoids have been isolated from H. angustifolia
root (Chen et al. 1994 2006), and some of them have
been proven to have antidiabetic activity. In this study,
the total flavonoid content in the ethanol extract of H.
angustifolia root was also determined which is much
higher than those of mulberry leaves exhibiting anti-
diabetic activity (Wanyo et al. 2011). Further studies are
necessary for the characterization and quantification of
the active ingredients in the ethanol extract of H.
angustifolia root.
Conclusion
Based on the results from cell-based bioassays and
animal-based assays, it can be concluded that the ethanol
extract from H. angustifolia root possesses potent
antidiabetic activity.
Disclosure statement
The authors confirmed that there is no conflict of
interests in the present study.
Funding information
This study was funded by Taisei Kogyo Co., Ltd (No.
CDE26014).
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