Journal of Ethnopharmacology 131 (2010) 140–145

Contents lists available at ScienceDirect

Journal of Ethnopharmacology
journal homepage: www.elsevier.com/locate/jethpharm

Anti-hyperglycemic activity of the leaves of Tetracera scandens Linn. Merr.

(Dilleniaceae) in alloxan induced diabetic rats
Abdulrashid Umar a , Qamar U. Ahmed a,∗ , Bala Y. Muhammad b ,
Bashar Bello S. Dogarai a , Siti Zaiton Bt. Mat Soad a
a
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, International Islamic University Malaysia (IIUM), 25200 Kuantan, Pahang DM, Malaysia
b
Department of Basic Medical Sciences, Faculty of Pharmacy, International Islamic University Malaysia (IIUM), 25200 Kuantan, Pahang DM, Malaysia

a r t i c l e i n f o a b s t r a c t

Article history: Aim of the study: The present study was aimed to investigate the anti-diabetic potential of the leaves of
Received 2 April 2010 Tetracera scandens Linn. Merr. (Dilleniaceae) in vivo with regard to prove its efficacy by local herbalists
Received in revised form 10 June 2010 in the treatment of diabetes frailties.
Accepted 11 June 2010
Materials and methods: Crude aqueous (AQ) and methanol (MEOH) extracts of the leaves of T. scandens
Available online 18 June 2010
L. were administered to both normal and alloxan induced diabetic male albino rats (Wistar strain). The
blood glucose levels were measured at 0, 2, 4, 6 and 8 h after oral administration of AQ and MEOH
Keywords:
extracts.
Tetracera scandens L.
Dilleniaceae
Results: Significant reduction in glucose was observed in fasting blood glucose levels in the treated diabetic
Alloxan induced diabetic rats rats without causing any hypoglycemic effect compared to normal rats. Both polar extracts of the leaves
Anti-hyperglycemic activity in vivo of T. scandens L. exhibited significant anti-hyperglycemic activity at different doses and intervals. The
highest anti-hyperglycemic effect (62.5%) was observed by the AQ extract at 0.25 g/kg body weight (b.w.)
and MEOH extract (36.5%) at 0.5 g/kg b.w. after 8 h. The significant anti-hyperglycemic activity was found
to be comparable with a known oral synthetic hypoglycemic drug, glibenclamide 0.25 mg/kg b.w.
Conclusion: This study provides scientific evidence that the leaves of T. scandens L. have anti-diabetic
efficacy and justifies its utility by local herbalists. However, more experiments at the clinical levels are
required to confirm the utility of this plant by traditional practitioners in the management of diabetes
mellitus.
© 2010 Elsevier Ireland Ltd. All rights reserved.

1. Introduction ation is given to the disease by health care managements both at

Diabetes mellitus is a chronic metabolic disorder which is char-
acterized by hyperglycemia (excessive hepatic glycogenolysis and
gluconeogenesis) resulting from the deficiency in the production of
insulin by the pancreas or its action. The etiology of type-1 diabetes
is the absolute deficiency of insulin secretion, while that of type-2 is
a combination of insulin resistance with inadequate compensatory
insulin-secretory response. Patients with diabetes experience sig-
nificant morbidity and mortality from microvascular (retinopathy,
neuropathy, and nephropathy) and macrovascular complications
(heart attack, stroke and peripheral vascular disease) (Altan, 2003).
The cost of treating diabetes and associated complications exceeds
$100 billion per year. The complications are far less common and
less severe in people who have well-controlled blood sugar levels.
Due to the multiplication of diabetics worldwide, a great consider-
∗ Corresponding author. Tel.: +60 9 5716400; fax: +60 9 5716775.
E-mail addresses: quahmed@iiu.edu.my, qamaruahmed@yahoo.com
(Q.U. Ahmed).
national and international levels. Over the past ten years, the num-
ber of diabetics has risen by 5% par annum (p.a.) to approximately
250 million and has been projected to become one of the world’s
major killers within next 25 years (Zimmet et al., 2001).
The disease affects all regions of the world but the share of dia-
betics in the overall population is particularly high in the eastern
Mediterranean countries and the Middle East (9%), in North Amer-
ica (8%) and in Europe (7%). Up until 2025 the WDF (World Diabetes
Foundation) expects the number of diabetics to increase by 2.5%
per year to about 380 million (Wild et al., 2004). Hence, the neces-
sity for developing newer drug therapies to prevent the burden
of complications associated with such disease. Natural products
and their derivatives have historically been precious as a source of
therapeutic agents. There is now a greater interest in the scientific
community to evaluate both crude and isolated natural products in
experimental studies and traditional medicines have always been
proved to be a fruitful source of future drugs to counteract any
disease including insulin resistance, consistent with a resurgence
of interest in drug discovery from natural products (Koehn and
Carter, 2005; Bnouham et al., 2006; Frode and Medeiros, 2008). The

0378-8741/$ – see front matter © 2010 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.jep.2010.06.016
 A. Umar et al. / Journal of Ethnopharmacology 131 (2010) 140–145
chosen plant given below could be a potential candidate for this
aim.
Tetracera scandens (Linn.) Merr. (T. scandens L.) (Family: Dil-
leniaceae) (Synonyms: Delima sarmentosa L., Tragia scandens L.,
Tetracera monocarpa Blanco, Tetracera hebecarpa (DC.) Boerl.) (local
Malaysian names-mempelas kasar, palas, pampan, and stone leaf)
is a climbing vine growing from 3 to 5 m or more in length and grows
widely in India, southern China, Indonesia, Myanmar, Philippines,
Thailand, Vietnam and Malaysia.
Different parts (leaves, stems and roots) of T. scandens L. are
used in folk remedies by various indigenous people in different
countries for the treatment of rheumatism, lowering hypertension,
lowering blood pressure, inflammatory diseases, hepatitis, internal
pains, urinary disorders, dysentery, child birth, sore throat, gout and
diabetes infirmities. Traditionally, in different regions of Malaysia,
the leaves of T. scandens L. are applied to boils to ripen them, juice
is generally taken to treat internal pains, a decoction of the plant
is administered after childbirth, the roots are used as an astrin-
gent in diarrhea and a traditional ingredient in a mixture against
burns, droplets of water/sap from freshly cut stems are used for eye
irritation, juice gathered by smashing the stem is taken to reduce
body heat and roots are ground and the juice of it is applied to
mouth ulcers. In Indonesia, the finely crushed young shoots are
made into a poultice and put on bites of poisonous snakes. The sap
of the stem is drunk as a cough medicine. In Philippines, an infu-
sion of the stem is drunk against haemoptysis in tuberculosis. It is
used as a gargle against thrush. Externally the infusion is applied
to a sore throat, the action being due to the large amount of tan-
nins, it contains. In Cambodia, the stem is used as a diuretic, and
in combination with other plants in oedemas of hepatic and renal
origin. In Vietnam, root and stem are used in hepatitis, gout and
inflammation. In India, juice of aerial portion is given orally once a
day in burning sensation during urination (Tawan, 2001; Werner,
2002; Nguyen et al., 2004; Purkayastha et al., 2007; Myung et al.,
2009).
The polar extracts of T. scandens L. have been reported to
exhibit potential therapeutic Xanthine oxidase (XO) inhibitory
activity in a concentration-dependent manner in vitro (Nguyen et
al., 2004). Five iso-flavonoids namely: genistein and its derivatives
3,5-diprenylgenistein, 6,8-diprenylgenistein, derrone and alpinum
isoflavone, isolated from the leaves of T. scandens L., have been
shown to exert significant glucose uptake effect in basal and
insulin-stimulated L6 Myotubes in vitro suggesting its great poten-
tial in the management of diabetes (Myung et al., 2009). However,
no scientific report of this plant in vivo has ever been recorded
or mentioned in the literature showing an anti-diabetic effi-
cacy of the leaves of T. scandens L. with respect to confirm its
utility in folkloric medicine by the local herbalists in Malaysia,
where leaves are used in herbal medicine preparations in diabetes
frailties.
Significant glucose uptake activity of the polyphenolic (iso-
flavonoids) compounds of T. scandens L. in basal and insulin-
stimulated L6 Myotubes in vitro and its usage by the local
traditional practitioners in different formulations to treat various
ailments including diabetes related complications in Malaysia have
prompted our current research interest in evaluating the efficacy of
the leaves of T. scandens L. in the management of diabetes mellitus
using animal model to confirm its effectiveness.
2. Materials and methods
2.1. Collection of plant material
Fresh leaves (10 kg) of T. scandens L. were collected from Taman
Pertanian, Kuantan, Indera Mahkota, 25200 Kuantan, Pahang Darul
141
Makmur, Malaysia, in October 2008. The plant was identified by Dr.
Richard Chung Cheng Kong (Taxonomist), F.R.I.M., K.L., Malaysia.
The voucher specimen (NMPC-Q24) has been deposited in the
Herbarium, Kulliyyah of Pharmacy, IIUM, Kuantan, Pahang DM,
Malaysia for future references.
2.2. Preparation of polar extracts of the leaves of T. scandens L.
The authenticated fresh leaves were dried in a laboratory dryer
for a temperature range (30–40 ◦ C) and were pulverized to a coarse
powdered form using Universal Cutting Mill, and the grinded mate-
rial was used for the extraction processes.
2.2.1. Aqueous (AQ) extract of the leaves of T. scandens L.
0.5 kg powdered (pulverized) material was soaked in 2l of ster-
ile double-distilled water in a tightly closed round bottom flask at
room temperature for a period of 24 h and filtered through Buch-
ner funnel. The whole process was repeated three times and further
heated at 95 ◦ C on water bath for 1 h to ensure maximum yield of
water soluble compounds. The extract was freeze-dried to give a
final yield of 50 g AQ extract (10%).
2.2.2. Methanol (MEOH) extract of the leaves of T. scandens L.
1.5 kg of powdered material was soaked in 3l analytical grade
methanol for 24 h, filtered through Buchner funnel, concentrated
using Buchi rotary evaporator and the recovered methanol was
again poured into the already extracted powder, filtered and con-
centrated. The entire process was repeated till the plant material
produced no coloration and eventually freeze-dried giving a final
yield of 205.4 g MEOH extract (13.69%).
2.3. Experimental animals
Adult male albino rats of the Wistar strain weighing 200–280 g,
between 2 and 3 mo. were used in this study. They were carefully
housed in an air conditioned animal house, maintained under stan-
dard condition of temperature 25 ± 5 ◦ C, relative humidity 35–60%
and 12 h light/dark cycles. All experimental subjects were allowed
to acclimatize laboratory ambiance for a period of 2 wk, fed with
standard pellet diet and tap water ad libitum, prior to the experi-
ment (Frode and Medeiros, 2008).
2.4. Acute toxicity study of AQ and MEOH extracts of the leaves of
T. scandens L.
A separate experiment was conducted to know whether any
toxic effect could be produced by AQ and MEOH extracts of the
leaves of T. scandens L. on normal rats. Normal healthy male rats
starved for 12 h were randomly divided into two groups (n = 10)
and were fed orally with both AQ and MEOH extracts separately
starting with the dose of 0.15, 0.48, 1.536, 4.915.2 and 5 g/kg
body weight (b.w.), respectively. Animals were dosed individu-
ally and observed continuously for 48 h and their behavioural and
neurological changes were vigilantly observed for sign of acute tox-
icity by using up and down method (OECD, 2008). All rats were
allowed to pellet diet and tap water ad libitum, and the mor-
tality caused by the extracts within this period of time was also
observed.
2.5. Induction of diabetes
Diabetes mellitus was induced in the rats by single intraperi-
toneal injection of 160 mg/kg b.w. of freshly prepared alloxan
monohydrate in normal saline. In order to prevent fatal hypo-
glycemia due to massive pancreatic insulin release, rats were
142 A. Umar et al. / Journal of Ethnopharmacology 131 (2010) 140–145
treated with 20% glucose solution intraperitoneally after 6 h fol-
lowed by 5% glucose solution bottles in their cages for a period
of 24 h. After one wk, the animals showing blood glucose level
>13.8 mmol/l were considered diabetic and used for the study. All
experiments were conducted under strict observance of animal
ethics guidelines and with permission from the ethics committee
of International Islamic University Malaysia (IIUM) (approval no.
IIUM/305/20/4/10/3/2009), Faculty of Medicine, Kuantan Campus,
IIUM, Malaysia.
2.6. Experimental design
Experiment consisted of 132 male albino rats of the Wistar
strain, divided equally (n = 6) among the AQ and MEOH extracts
treatment groups, and fasted overnight. The MEOH extract pre-
pared in the form of emulsion (oil:water 6:4) and AQ extract in
sterile double-distilled water were administered to the experimen-
tal animals by gastric intubation using forced feeding needle. Blood
samples from the tail vein were collected for the measurement
of blood glucose at 0 (baseline), 2, 4, 6 and 8 h after administer-
ing the extracts. Blood glucose was measured using one touch
Ultra glucometer and the results were compared with those of
6th group treated with glibenclamide (0.25 mg/kg b.w.) a standard
oral hypoglycemic agent (Ji et al., 2006). The rats were randomly
distributed in eleven groups separately as follows both for the
aqueous (AQ) as well as for the methanol (MEOH) extracts sepa-
rately, viz., untreated diabetic control rats (group 1 (AQ extract), 1
(MEOH extract)), diabetic rats receiving the AQ or MEOH extracts
at 0.25 g/kg b.w. (group 2, 2 ), 0.5 g/kg b.w. (group 3, 3 ), 1 g/kg
b.w. (group 4, 4 ), 2 g/kg b.w. (group 5, 5 ), diabetic rats treated
with glibenclamide (GLBC) 0.25 mg/kg b.w. (group 6, 6 ) normal
control rats (group 7, 7 ), normal rats receiving the AQ or MEOH
extracts at 0.25 g/kg b.w. (group 8, 8 ), 0.5 g/kg b.w. (group 9, 9 ),
1 g/kg b.w. (group 10, 10 ) and 2 g/kg b.w. (group 11, 11 ). In case
of MEOH extract, group 1 and 7 were administered the emulsion
(oil:water 6:4) only and in case of AQ extract, group 1 and 7 were
administered sterile double-distilled water to confirm any unto-
ward, anti-diabetic or cumulative effect by both emulsion as well
as sterile double-distilled water in the rats, respectively (Rao et al.,
2003).
2.7. Phytochemical characterization of AQ and MEOH extracts of
the leaves of T. scandens L.
Total contents of phenols in AQ and MEOH extracts of the leaves
of T. scandens L. were determined by the Folin-Ciocalteu assay
(McDonald et al., 2001) with some modifications. Briefly, 0.25 ml
of plant extracts (0.1 mg/ml) were mixed with 2.5 ml of Folin-
Ciocalteu reagent (diluted 1:10 with distilled water) and allowed
to stand for 5 min at room temperature. Then, 2 ml of 1 M Na2 CO3
were added and the mixture was incubated at room temperature
for 2 h. Finally, total phenols were estimated at 765 nm using a
spectrophotometer (Jenway 6405 UV/vis Dunmow, Essex, UK). A
standard curve was prepared using gallic acid (0–250 mg/l). Total
phenol values were expressed as gallic acid equivalents (GAE) mg/g
of plant extracts.
Total flavonoids in AQ and MEOH extracts of the leaves of T.
scandens L. were estimated using the Dowd method as adapted by
Arvouet-Grand et al. (1994). Briefly, 2.5 ml of 2% AlCl3 in methanol
were mixed with 2.5 ml of plant extract (0.1 mg/ml). The mixture
was allowed to stand for 10 min at room temperature and the total
flavonoid content was determined by UV spectrophotometer at
415 nm using a quercetin (0–250 mg/l) standard curve. Flavonoids
contents were expressed as quercetin equivalents (QE) mg/g of
plant extract.
Thin layer chromatography (TLC) evaluation of AQ and MEOH
extracts of the leaves of T. scandens L. was carried out on silica gel
60 F254, 0.2 mm thickness aluminium plates, in Benzene:Acetone
(B:A) (9:1, 5:1 and 3:1), Toluene:Ethylformate:Formic acid
(T:E:F) (5:4:1), Benzene:Pyridine:Formic acid (B:P:F) (39:6:5),
Chloroform:MeOH:Formic acid (C:M:F) (90:05:0.6) and Ben-
zene:Acetone:Formic acid (B:A:F) (3:1:0.1), respectively to confirm
the presence of different class of phytochemicals by using different
class of selected reagents (Harbone, 1998).
2.8. Statistical analysis
The results are expressed as mean ± standard deviation (SD).
Data were evaluated using Repeated Measurement GLM ANOVA
(SPSS version 17.0). Values of p < 0.05 were considered signifi-
cant.
3. Results
3.1. Plant extraction and phytochemical characterization of AQ
and MEOH extracts of the leaves of T. scandens L.
0.5 kg leaves of T. scandens L. produced 50 g freeze-dried AQ
extract and 1.5 kg leaves yielded 205.4 g freeze-dried MEOH extract.
TLC examination of these extracts was characterized by analyzing
their contents of phenols, flavonoids, and alkaloids, the main com-
pounds reputed as to be responsible for the anti-hyperglycemic
properties of many anti-diabetic plants (Jung et al., 2006). The
chemical characterization of AQ extract resulted in 4.75 ± 0.02 GAE
mg/g of phenolic compounds, 5.77 ± 0.02 QE mg/g of flavonoids,
whereas the chemical characterization of MEOH extract was
7.26 ± 0.03 GAE mg/g of phenolic compounds, 6.34 ± 0.02 QE
mg/g of flavonoids. Phytochemical TLC analysis of MEOH and
AQ extracts of the leaves of T. scandens L. in BA, TEF, BPF, CMF
and BAF solvent systems vividly showed the presence of more
than ten spots of phenolic compounds and six terpenoidal com-
pounds.
3.2. Determination of non-toxic concentrations of AQ and MEOH
extracts of the leaves of T. scandens L. for in vivo assays
Acute toxicity studies explicitly revealed the non-toxic safe
nature of both AQ as well as MEOH extracts of the leaves of T. scan-
dens L. Experiment was carried out on normal healthy male rats. No
mortality was observed in the extracts treated rats and behaviour
of the treated rats also appeared normal in most of the animals.
Neither lethality nor any toxic reaction found at any dose selected
until the end of the study.
3.3. Determination of anti-diabetic activity of AQ and MEOH
extracts of the leaves of T. scandens L. in vivo
Administration of single intraperitoneal injection of 160 mg/kg
b.w. of freshly prepared alloxan monohydrate produced diabetes
in rats after a week. The effect of different doses of both AQ and
MEOH extracts of the leaves of T. scandens L. on fasting blood glu-
cose levels in normal and alloxan induced diabetic male albino
rats (Wistar strain) are summarized in Tables 1 and 2, respec-
tively. The fasting blood glucose levels in alloxan induced diabetic
rats were in the range of 13.8–30.9 mmol/l. The fasting blood glu-
cose levels of diabetic untreated rats (group 1) were significantly
higher than that of normal untreated rats (group 7) and the treat-
ment with different doses of both AQ and MEOH extracts did
not result hypoglycemia in normal rats (Tables 1 and 2). How-
ever, all doses of AQ extract showed a significant reduction in
 A. Umar et al. / Journal of Ethnopharmacology 131 (2010) 140–145 143

Table 1
Effect of different doses of aqueous (AQ) extract of the leaves of T. scandens L. on blood glucose levels (mmol/l) in normal and diabetic rats at different intervals (h). The values
represent the standard error of mean (S.E.M.).

AQ extract groups/dose 0h 2h 4h 6h 8h

1. Diabetic control rats 27.9 ± 5.4 28.5 ± 5.4 28.9 ± 5.8 27.8 ± 5.6 28.1 ± 5.9
2. Diabetic rats/0.25 g/kg b.w. 25.3 ± 4.1 18.7 ± 1.6* 14.8 ± 2.4** 13.1 ± 2.4** 9.5 ± 4.2**
3. Diabetic rats/0.5 g/kg b.w. 26.9 ± 3.1 25.6 ± 4.8 18.1 ± 6.5** 16.3 ± 6.9** 12.8 ± 7.6*
4. Diabetic rats/1 g/kg b.w. 26.4 ± 5.9 21.3 ± 5.6 17.8 ± 5.7** 15.4 ± 6.6** 12.8 ± 5.2**
5. Diabetic rats/2 g/kg b.w. 26.5 ± 4.1 22.3 ± 3.0 21.3 ± 4.5 19.6 ± 2.9* 15.5 ± 2.0**
6. Diabetic rats/GLBC 0.25 mg/kg b.w. 26.3 ± 4.1 20.8 ± 4.5 14.4 ± 5.6** 13.1 ± 4.6** 10.9 ± 4.6**
7. Normal control rats 3.96 ± 0.2 3.6 ± 0.2 3.6 ± 0.8 3.5 ± 0.3 3.5 ± 0.2
8. Normal rats/0.25 g/kg b.w. 4.9 ± 1.2 4.6 ± 1.0 4.1 ± 0.8 4.0 ± 0.7 3.6 ± 0.7
9. Normal rats/0.5 g/kg b.w. 4.7 ± 0.9 4.6 ± 0.7 4.5 ± 0.6 4.0 ± 0.6 3.4 ± 0.6
10. Normal rats/1 g/kg b.w. 4.8 ± 0.5 5.9 ± 1.6 5.3 ± 1.0 4.7 ± 0.7 4.6 ± 1.0
11. Normal rats/2 g/kg b.w. 4.0 ± 0.9 3.6 ± 0.2 4.3 ± 1.0 4.2 ± 0.4 4.0 ± 0.4

GLBC: glibenclamide.
*
p < 0.05.
**
p < 0.005.

Table 2
Effect of different doses of methanol (MEOH) extract of the leaves of T. scandens L. on blood glucose levels (mmol/l) in normal and diabetic rats at different intervals (h). The
values represent the standard error of mean (S.E.M.).

MEOH extract groups/dose 0h 2h 4h 6h 8h


1 . Diabetic control rats 24.9 ± 6.0 25.4 ± 7.2 23.7 ± 6.4 24.4 ± 7.2 24.6 ± 7.4
2 . Diabetic rats/0.25 g/kg b.w. 30.9 ± 4.5 28.3 ± 3.4 25.8 ± 3.6 23.6 ± 3.3 19.8 ± 2.5
3 . Diabetic rats/0.5 g/kg b.w. 22.2 ± 3.2 20.2 ± 4.1 18.4 ± 5.5 16.4 ± 6.2* 14.1 ± 5.9**
4 . Diabetic rats/1 g/kg b.w. 22.9 ± 4.3 20.7 ± 3.6 18.7 ± 2.4 17.3 ± 4.2* 15.6 ± 6.4*
5 . Diabetic rats/2 g/kg b.w. 26.9 ± 3.3 24.7 ± 3.9 22.8 ± 5.2 20.9 ± 6.2 18.8 ± 5.5
6 . Diabetic rats/GLBC 0.25 mg/kg b.w. 24.6 ± 4.1 22.8 ± 5.2 22.1 ± 4.5 17.4 ± 4.2* 14.6 ± 3.8*
7 . Normal control rats 5.8 ± 0.9 5.3 ± 0.6 4.8 ± 0.7 4.7 ± 0.7 4.8 ± 0.3
8 . Normal rats/0.25 g/kg b.w. 5.7 ± 0.4 4.9 ± 0.4 4.8 ± 1.0 5.1 ± 0.9 5.3 ± 0.7
9 . Normal rats/0.5 g/kg b.w. 6.0 ± 0.7 5.0 ± 0.3 4.6 ± 0.6 4.3 ± 0.8 4.7 ± 0.9
10 . Normal rats/1 g/kg b.w. 5.9 ± 0.4 5.9 ± 0.5 5.9 ± 0.6 6.2 ± 0.8 6.1 ± 0.5
11 . Normal rats/2 g/kg b.w. 6.4 ± 0.5 5.9 ± 0.8 6.2 ± 0.5 5.8 ± 0.8 6.4 ± 1.4

GLBC: glibenclamide.
*
p < 0.05.
**
p < 0.005.

blood glucose level (i.e., anti-hyperglycemic activity) in the dia-
betic rats at 4, 6 and 8 h post-treatment, respectively, while the
highest percentage in the blood glucose fall was observed with
0.25 g/kg b.w. at 8 h post-treatment (62.5%) (P < 0.005) (Fig. 1).
0.5 and 1 g/kg b.w. doses of MEOH extract produced signifi-
cant anti-hyperglycemic activity (27.1% and 36.5%, respectively)
in the diabetic rats at 6 and 8 h post-treatment (Fig. 2). Treat-
ment with glibenclamide at a dose of 0.25 mg/kg b.w. in the
6th group of diabetic rats resulted 58% (Fig. 1) and 40% (Fig. 2)
fall of blood glucose level at 8 h in the AQ and MEOH extracts
groups, respectively. The anti-hyperglycemic activity of AQ extract
at the dose of 0.25 g/kg b.w. in diabetic rats was found to be
higher (26.1% and 62.5%) at 2 and 8 h post-treatment, respectively
than that of the oral hypoglycemic agent glibenclamide (GLBC)
(20.9% and 58%, respectively) (Fig. 1). The anti-hyperglycemic activ-
ity of MEOH extract at all the doses in diabetic rats was found
to be higher at 2 and 4 h post-treatment than that of the oral
hypoglycemic agent, glibenclamide (GLBC) (Fig. 2). The highest
anti-hyperglycemic effect (62.5%) (Fig. 1) was observed by the AQ
extract at 0.25 g/kg b.w. and MEOH extract (36.5%) (Fig. 2) at 0.5 g/kg
b.w. after 8 h, respectively. The significant anti-hyperglycemic
activity was found to be comparable with a known oral synthetic
hypoglycemic drug, glibenclamide (GLBC) (Tables 1 and 2). Differ-
ent doses of AQ and MEOH extracts of the leaves of T. scandens L.

Fig. 1. Percentage fall of blood glucose of different doses of aqueous (AQ) extract of Fig. 2. Percentage fall of blood glucose of different doses of methanol (MEOH)
the leaves of T. scandens L. in diabetic rats. (mmol/l); GLBC: glibenclamide. extract of the leaves of T. scandens L. in diabetic rats. (mmol/l); GLBC: glibenclamide.
144 A. Umar et al. / Journal of Ethnopharmacology 131 (2010) 140–145
produced a significant fall in the blood glucose levels of diabetic rats
(anti-hyperglycemic effect) (Tables 1 and 2) and the same doses
did not exhibit any hypoglycemic effect in normal non-diabetic
rats.
4. Discussion and conclusion
Presently, different synthetic drugs viz., biguanides, thiazo-
lidinediones, sulphonylureas, diphenylalanine derivatives, megli-
tinides and ␣-glucosidase inhibitors in addition to insulin, are
widely used in the management of diabetes all over the world. How-
ever, due to untoward side effects, the efficacies of these drugs are
quite controversial and there is a strong demand for new but safe
drugs for the treatment of diabetes efficaciously (Thirunavukkarasu
et al., 2003). Hence, plants have been suggested as a rich, as yet
unexplored source of potentially useful anti-diabetic drugs (Koehn
and Carter, 2005; Bnouham et al., 2006; Frode and Medeiros, 2008).
However, only a few have been subjected to detailed scientific
exploration due to a lack of mechanism-based available in vitro
assays (Saxena and Vikram, 2004).
A number of experiments have vividly shown the beneficial
effects of medicinal plants in the management of diabetes mel-
litus. Numerous mechanisms of actions have been proposed for
these plant extracts. Some reports have linked their effects to
the activity of pancreatic ␤-cells (synthesis, release, cell regenera-
tion/revitalization) (Yi et al., 2009) or the increase in the inhibitory
effect against insulinase and the increase of the insulin sensitiv-
ity or the insulin-like activity of the plant extracts (Meng et al.,
2009; Angel et al., 2010). Others have suggested that the mecha-
nisms may involve improved glucose homeostasis (Ahmad et al.,
2000), increase of peripheral utilization of glucose, increase of syn-
thesis of hepatic glycogen (Yuan et al., 1998) and/or decrease of
glycogenolysis acting on enzymes (El-Missiry and El Gindy, 2000)
inhibition of intestinal glucose absorption (Nicola et al., 1996),
reduction of glycaemic index of carbohydrates (Frati Munari et
al., 1998) and reduction of the effect of glutathione (Raza et al.,
1996).
In our present study, different doses of AQ extract of the leaves
of T. scandens L. produced a significant fall in the blood glucose
levels of diabetic rats and the same doses did not exhibit any
hypoglycemic effect in normal non-diabetic rats. The diminished
anti-hyperglycemic activity of MEOH extract at the dose higher
than 1 g/kg b.w. could be due to the presence of other antago-
nistic components in the MeOH extract (Prince et al., 1999), in
contrast, the AQ extract produced significant and consistent anti-
hyperglycemic effect in the diabetic rats. Although, both the AQ
and MEOH extracts of the leaves of T. scandens L. explicitly exhib-
ited significant anti-hyperglycemic activity in the diabetic rats, the
AQ extract proved more potent in comparison, however, there was
no hypoglycemic activity observed with both extracts in normal
rats.
Flavonoids, terpenoids, steroids, and alkaloids are known to
be bioactive anti-diabetic agents (Jung et al., 2006). Moreover,
flavonoids are known to regenerate the damaged ␤-cells in the
alloxan induced diabetic rats and are considered effective anti-
hyperglycemic agents (Rao et al., 2003). It has already been
investigated in vitro that genistein and its derivatives: 3,5-diprenyl
genistein, 6,8-diprenylgenistein, derrone, and alpinumisoflavone
isolated from the leaves of T. scandens L. exhibited significant glu-
cose uptake effect in basal and insulin stimulated L6 myotube in
vitro suggesting its role in the management of diabetes (Myung et
al., 2009). Presence of greater amount of flavonoids content in the
leaves of T. scandens L. could be responsible for unleashing the sig-
nificant anti-hyperglycemic activity of AQ as well as MEOH extracts
in vivo.
Many new bioactive principles isolated from plants having anti-
hyperglycemic effects have shown anti-diabetic activity equal and
even more potent than known oral hypoglycemic agents such as
daonil, tolbutamide and chlorpropamide. However, many other
active agents obtained from plants have not been well character-
ized and documented. More investigations and sincere efforts are
still sought to evaluate the precise mechanism of action of medici-
nal plants with anti-diabetic effect at the molecular level (Bnouham
et al., 2006).
Based on this study we can conclusively affirm that the polar
extracts (AQ and MEOH extracts) of the leaves of T. scandens L.
exhibited significant anti-hyperglycemic effect in alloxan induced
diabetic rats without showing any hypoglycemic effect and justi-
fies its utility by the local traditional practitioners in the treatment
of diabetes infirmities in Malaysia; however, further chemical and
pharmacological studies at the molecular and clinical levels are still
warranted to identify the anti-diabetic agent responsible for elic-
iting aforementioned anti-hyperglycemic activity and its precise
mechanism of action.
Conflict of interest statement
None.
Acknowledgements
We particularly wish to thank the traditional practitioners and
field supervisors who helped to collect the plant material. We also
thank the botanists of F.R.I.M. (Kuala Lumpur, Malaysia) for the
botanical determination of the plant. All authors are also extremely
indebted to Research Management Center, IIUM for furnishing
grant from the endowment fund-B (EDW-B0804-124) to accom-
plish this work.
References
Ahmad, M., Akhtar, M.S., Malik, T., Gilani, A.H., 2000. Hypoglycaemic action of
the flavonoid fraction of Cuminum nigrum seeds. Phytotherapy Research 14,
103–106.
Altan, V.M., 2003. The pharmacology of diabetic complications. Current Medicinal
Chemistry 10, 1317–1327.
Angel, J.A.C., Rocio, Z.B., José, R.Y., Paul, C.L., Maricela, G.S., Luis, A.S.O., 2010. The
antidiabetic plants Tecoma stans (L.) Juss. ex Kunth (Bignoniaceae) and Teucrium
cubense Jacq (Lamiaceae) induce the incorporation of glucose in insulin-sensitive
and insulin-resistant murine and human adipocytes. Journal of Ethnopharma-
cology 127, 1–6.
Arvouet-Grand, A., Vennat, B., Pourrat, A., Legret, P., 1994. Standardisation d’un
extrait de propolis et identification des principaux constituants. Journal de Phar-
macie de-Belgique 49, 462–468.
Bnouham, M., Ziyyat, A., Mekhfi, H., Tahri, A., Legssyer, A., 2006. Medicinal plants
with potential antidiabetic activity—a review of ten years of herbal medicine
research (1990–2000). International Journal of Diabetes & Metabolism 14,
1–25.
El-Missiry, M.A., El Gindy, A.M., 2000. Amelioration of alloxan induced diabetes mel-
litus and oxidative stress in rats by oil of Eruca sativa seeds. Annals of Nutrition
& Metabolism 44, 97–100.
Frati Munari, A.C., Benitez Pinto, W., Raul Ariza, A.C., Casarrubias, M., 1998. Lowering
glycaemic index of food by acarbose and Plantago psyllium mucilage. Archives
of Medical Research 29, 137–141.
Frode, T.S., Medeiros, Y.S., 2008. Animal model to test drugs with potential antidia-
betic activity. Journal of Ethnopharmacology 115, 173–183.
Harbone, J.B., 1998. Phytochemical Methods: A Guide to Modern Techniques of Plant
Analysis, third edition. Thompson Publishing IT, 5–21.
Ji, S.K., Jung, B.J., Chang, W.C., Sei, C.K., 2006. Hypoglycemic and antihyperlipidemic
effect of four Korean medicinal plants in alloxan induced diabetic rats. American
Journal of Biochemistry and Biotechnology 2, 154–160.
Jung, M., Park, M., Lee, H.C., Kang, Y.H., Kang, E.S., Kim, S.K., 2006. Antidi-
abetic agents from medicinal plants. Current Medicinal Chemistry 13,
1203–1218.
Koehn, F.E., Carter, G.T., 2005. The evolving role of natural products in drug discovery.
Nature Reviews Drug Discovery 4, 206–220.
McDonald, S., Prenzler, P.D., Autolovich, M., Robards, K., 2001. Phenolic content and
antioxidant activity of olive extracts. Food Chemistry 73, 73–84.
Meng, J.L., Yerra, K.R., Keru, C., Yi, C.L., Yew, M.T., 2009. Effect of flavonol glycosides
from Cinnamomum osmophloeum leaves on adiponectin secretion and phospho-
 A. Umar et al. / Journal of Ethnopharmacology 131 (2010) 140–145
rylation of insulin receptor- in 3T3-L1 adipocytes. Journal of Ethnopharmacology
126, 79–85.
Myung, S.L., Chung, H.K., Duc, M.H., Bo, Y.K., Cheon, B.S., Mee, R.K., Jong, S.A., 2009.
Genistein derivatives from Tetracera scandens stimulate glucose uptake in L6
myotubes. Biological & Pharmaceutical Bulletin 32, 504–508.
Nguyen, M.T., Awale, S., Tezuka, Y., Tran, Q.L., Watanabe, H., Kadota, S., 2004. Xan-
thine oxidase inhibitory activity of Vietnamese medicinal plants. Biological &
Pharmaceutical Bulletin 27, 1414–1421.
Nicola, W.G., Ibrahim, K.M., Mikhail, T.H., Girgis, R.B., Khadr, M.E., 1996. Role of the
hypoglycaemic plant extract Cleome droserifolia in improving glucose and lipid
metabolism and its relation to insulin resistance in fatty liver. Bull Chim Farm
135, 507–517.
Organization for Economic Co-operation and Development (OECD). OECD guidelines
for the testing of chemicals. Test No. 425: Acute Oral Toxicity: Up-and-Down Pro-
cedure. October 2008. (Available at http://lysander.sourceoecd.org/ (Accessed
on 21.3.09).
Prince, P.S.M., Menon, V.P., Gunasekharan, G., 1999. Hypolipidaemic action of
Tinospora cardifolia roots in alloxan diabetic rats. Journal of Ethnopharmacology
64, 53–57.
Purkayastha, J., Dutta, M., Nath, S.C., 2007. Ethnomedicinal plants from Dibru-
Saikhowa biosphere reserve, Assam. Indian Journal of Traditional Knowledge
6, 477–480.
Rao, B.K., Sudarshan, P.R., Rajasekhar, M.D., Nagaraju, N., Roa, C.A., 2003. Antidiabetic
activity of Terminalia pallida fruit in alloxan induced diabetic rats. Journal of
Ethnopharmacology 85, 169–172.
145
Raza, H., Ahmed, I., Lakhani, M.S., Sharma, A.K., Pallot, D., Montague, W.,
1996. Effect of bitter melon (Momordica charantia) fruit juice on the
hepatic cytochrome P450-dependent monooxygenase and glutathione S-
transferase in streptozotocin-induced diabetic rats. Biochemical Pharmacology
52, 1639–1642.
Saxena, A., Vikram, N.K., 2004. Role of selected Indian plants in management of type
2 diabetes. Journal of Alternative and Complementary Medicine 10, 369–378.
Tawan, C.S., 2001. Tetracera scandens (L.) Merr. In: van Valkenburg, J.L.C.H., Bun-
yapraphatsara, N. (Eds.), Plant Resources of South-East Asia No. 12 (2): Medicinal
and Poisonous Plants 2. Backhuys Publisher, Leiden, The Netherlands, p. 543.
Thirunavukkarasu, V., Anuradha, C.V., Viswanathan, P., 2003. Protective effect of
fenugreek (Trigonella foenum graecum) seeds in experimental ethanol toxicity.
Phytotherapy Research 17, 737–743.
Werner, R., 2002. Medicines in Malay Villages: Malay Culture of Healing, vol. 2.
University of Malaya Press, Kuala Lumpur, Malaysia, pp. 82.
Wild, S., Roglic, G., Green, A., Sicree, R., King, H., 2004. Global prevalence of diabetes:
estimates for 2000 and projections for 2030. Diabetes Care 27, 1047–1053.
Yi, J.H., Tsung, H.L., Cicero, L.T.C., Yuh, T.H., Wen, C.Y., 2009. Anti-hyperglycemic
effects and mechanism of Bidens pilosa water extract. Journal of Ethnopharma-
cology 122, 379–383.
Yuan, Z., He, P., Cui, J., Takeuchi, H., 1998. Hypoglycaemic effect of water-soluble
polysaccharide from Auricularia auricula-judae Quel. on genetically diabetic KK-
Ay mice. Bioscience, Biotechnology and Biochemistry 62, 1898–1903.
Zimmet, P.Z., Alberti, K.G.M.M., Shaw, J., 2001. Global and societal implications of
the diabetes epidemic. Nature 414, 782–787.