Journal of Ethnopharmacology 70 (2000) 309 314

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Short communication

Effect of Punica granatum Linn. (flowers) on blood glucose

level in normal and alloxan-induced diabetic rats
M.A. Jafri a,*, M. Aslam a, Kalim Javed a, Surender Singh b
a

Department of Ilmul Ad6ia, Faculty of Medicine (Unani ), Jamia Hamdard, New Delhi-110 062, India
b
Department of Pharmacology, College of Pharmacy, Pushp Vihar, New Delhi, India
Received 28 April 1999; received in revised form 30 June 1999; accepted 27 September 1999

Abstract
Gulnar farsi, male abortive flowers of Punica granatum L., are used for the treatment of diabetes mellitus in Unani
medicine. Oral administration of its aqueous-ethanolic (50%, v/v) extract led to significant blood glucose lowering
effect in normal, glucose-fed hyperglycaemic and alloxan-induced diabetic rats. This effect of the extract was
maximum at 400 mg/kg, b.w. 2000 Elsevier Science Ireland Ltd. All rights reserved.
Keywords: Alloxan; Antihyperglycaemic; Diabetes; Gulnar; Hypoglycaemic; Punica granatum L.

1. Introduction
Punica granatum Linn. (Punicaceae) is a shrub
or small tree and considered to be a native of Iran
and Afghanistan. It is also found growing wild in
the warm valleys and outer hills of the Himalayas,
and is cultivated throughout India (Satyavati et
al., 1978). Gulnar (flower of P. granatum L.) has
been known for a long time in Unani literature as
an astringent, haemostatic, and as a remedy for
diabetes (Jurjani, 1878; Majoosi, 1889). The root
bark as well as the stem bark of the plant is
astringent and also used as anthelmintic specifically against tapeworms. The rind is valued as an
* Corresponding author.

astringent in diarrhoea and dysentery. The juice

of the leaves and young fruits and the decoction
of the bark are used in dysentery. The powdered
flower buds are useful in bronchitis. The seeds are
considered to be stomachic and the pulp cardiac
and stomachic. The green leaves are made into a
paste and applied in conjunctivitis (Anonymous,
1969; Satyavati et al., 1978).
The biological activities, viz. antibacterial (Chopra et al., 1960; Trivedi and Kazmi, 1979), antifungal (Janardhanan et al., 1963; Charya et al.,
1979), anthelmintic (Prakash et al., 1980; Singhal,
1983) and antifertility (Dhawan and Saxena, 1958;
Gujral et al., 1960), of the various extracts of
different parts of this plant have also been reported. The extracts of root of P. granatum (Car-

0378-8741/00/$ - see front matter 2000 Elsevier Science Ireland Ltd. All rights reserved.
PII: S 0 3 7 8 - 8 7 4 1 ( 9 9 ) 0 0 1 7 0 - 1

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M.A. Jafri et al. / Journal of Ethnopharmacology 70 (2000) 309314

raz et al., 1978) and rind of this plant (Nogueira

and Pereira, 1984, 1986a,b; Zafar and Singh,
1990) have been reported to exert some sugar
lowering action in animals.
Since only the flowering part of the plant has
been recommended in Unani literature (Jurjani
1878; Majoosi, 1889) as a remedy for the treatment of diabetes, it was, therefore, considered
worthwhile to investigate the effect of the flowers
of P. granatum on blood glucose levels of glucosefed hyperglycaemic, alloxan-induced diabetic and
normal rats and to compare it with tolbutamide
as a reference standard.

2.4. Beha6ioural effect and toxicity

Albino mice (Swiss strain) were divided into
five groups of ten animals each. The test drug
extract, suspended in vehicle (Tween 80 in distilled water, 10% v/v) was administered in the
dose of 200, 400, 600, 800 and 1000 mg/kg i.p. in
a volume of 10 ml/kg to the animals of I, II, III,
IV and V groups, respectively. Then the mice were
observed continuously for 1 h, intermittently for 6
h and at the end of 24 h for any gross behavioural
changes and deaths.

2.5. Effect of the test drug on oral glucose

tolerance
2. Materials and methods

2.1. Plant material

The flowers of P. granatum were purchased
under the Unani name Gulnar farsi from Khari
Baoli, Delhi, and its identity was confirmed by the
department of Botany, Faculty of Science, Jamia
Hamdard, New Delhi. Its voucher specimen (MA
98 01) was deposited in the pharmacognosy section of the Department of Ilmul Advia, Faculty of
Medicine (Unani) Jamia Hamdard, New Delhi.

2.2. Preparation of plant extract

The flowers of P. granatum (200 g) were extracted by refluxing with aqueous-ethanol (50%
v/v, 1000 ml) on boiling water bath for 6 h. The
result was filtered and residue was re-extracted in
the same manner four times. All the filtrates were
combined together and recovery of the solvent
under reduced pressure yielded semisolid mass
(80.20 g).

2.3. Experimental animals

Adult albino rats (Wistar strain) of either sex
(120 220 g) were obtained from the Animal
House, Jamia Hamdard, New Delhi. Animals,
described as fasted, were deprived of food for at
least 16 h but allowed free access to water.

Fasted rats were divided into five groups of six

animals each. Group I, serving as control, received only vehicle (Tween 80 in distilled water,
10% v/v) orally in a volume of 10 ml/kg and
group II received tolbutamide (500 mg/kg, p.o.)
suspended in vehicle (10 ml/kg). A dose of 300,
400 and 500 mg/kg of the extract of the test drug
suspended in vehicle was administered orally in a
volume of 10 ml/kg to the animals of III, IV and
V groups, respectively. All the animals were given
glucose (2 g/kg, p.o.) 30 min after dosing. Blood
samples were collected from the retro-orbital
plexus just prior to and at 30 and 90 min after the
glucose loading and blood glucose levels were
measured.

2.6. Effect of the test drug on blood glucose

le6els in normal fasted rats
Fasted rats were divided into five groups of six
rats each. Group I received only vehicle (Tween
80 in distilled water, 10% v/v) orally in a volume
of 10 ml/kg and served as control. The extract of
the test drug, suspended in vehicle, was administered at the doses of 300, 400 and 500 mg/kg
orally in a volume of 10 ml/kg to the animals of
group III, IV and V, respectively. Group II received tolbutamide (500 mg/kg, p.o.) suspended in
vehicle (10 ml/kg). Blood samples were collected
from the retro-orbital plexus just prior to and at
1, 2 and 3 h after dosing.

M.A. Jafri et al. / Journal of Ethnopharmacology 70 (2000) 309314

2.7. Effect of the test drug on alloxan induced

diabetes in rats
Rats were made diabetic by a single intraperitoneal injection of alloxan monohydrate 5% (w/v)
in normal saline at a dose of 120 mg/kg, b.w.
(Khosla et al., 1995). Then 5 days later blood
samples were collected and glucose levels were
determined to confirm the development of diabetes. Of the animals receiving alloxan,  82%
were found to be diabetic (fasting blood glucose
259 292 mg/dl). The diabetic rats were divided
into five groups of six animals each. Group I
received only vehicle (Tween 80 in distilled water,
10 v/v) orally in a volume of 10 ml/kg. The test
drug extract, suspended in vehicle, was given in
the doses of 300, 400 and 500 mg/kg orally in a
volume of 10 ml/kg to groups III, IV and V,
respectively. Group II received tolbutamide (500
mg/kg, p.o.) suspended in vehicle (10 ml/kg).
Blood samples were collected just prior to and at
1 and 2 h after dosing.

2.8. Estimation of blood glucose le6el

In each case 200 ml of blood sample was collected and estimated for glucose by o-toluidine
method (Dubowski, 1962).

2.9. Data and statistical analysis

Data were expressed as mean9S.E.M., n = 6.
Statistical comparisons within the same group

311

were performed with Students t-test for paired

observations. One-way analysis of variance
(ANOVA) followed by the least significant difference post-test was used to identify significantly
different groups. PB0.05 indicates significant differences between group means.

3. Results

3.1. Beha6ioural effect and toxicity

The extract of the test drug (P. granatum) was
found to be safe for further biological studies as
no lethality was observed at 1000 mg/kg, i.p. in
mice.

3.2. Effect of the test drug on oral glucose

tolerance
The effects of various doses of the extract of P.
granatum on glucose tolerance are shown in Table
1. By 30 min after starting the glucose tolerance
test, blood glucose concentration doubled from its
initial level of control and then glycaemia started
to decrease gradually, without reaching its initial
value till the end of the studies, i.e. at the 90th
min. All the doses of the test drug extract were
found to be effective in depressing the peak value
of blood sugar. Administration of tolbutamide
(500 mg/kg) to glucose-fed rats induced time dependent hypoglycaemic effect.

Table 1
Effect of oral administration of different doses of P. granatum extract and tolbutamide on oral glucose tolerance in normal ratsa
Sampling time
(min)

0
30
90

Blood glucose level (mg/dl)

Vehicle (10% v/v Tween 80, 10
ml/kg)

Tolbutamide,
500 mg/kg

P. granatum,
300 mg/kg

P. granatum, 400
mg/kg

P. granatum, 500
mg/kg

67.369 1.69
133.00 91.65
100.33 91.65

63.509 1.95
56.839 0.86b,c
41.3391.65b,c

59.33 9 1.60
110.00 9 3.89b
90.16 93.32b

63.66 91.72
100.66 93.85b
81.00 94.28b

64.00 9 1.06
99.33 9 3.54b
80.16 92.94b

Values are expressed as mean 9S.E.M (n = 6).

Statistically significant difference to the vehicle.
c
Statistically significant difference to the corresponding zero time value.
a

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M.A. Jafri et al. / Journal of Ethnopharmacology 70 (2000) 309314

Table 2
Effect of oral administration of different doses of P. granatum extract and tolbutamide on the mean blood glucose level in normal
fasted ratsa
Sampling time
(h)

0
1
2
3

Blood glucose level (mg/dl)

Vehicle (10% v/v Tween 80, 10
ml/kg)

Tolbutamide,
500 mg/kg

P. granatum,
300 mg/kg

P. granatum, 400
mg/kg

P. granatum, 500
mg/kg

57.339 1.45
66.169 1.16
67.509 0.61
64.339 0.84

66.339 2.45
60.689 0.33b,c
46.0091.24b,c
40.169 1.11b,c

63.50 9 1.96
69.33 90.71
57.50 90.76b,c
68.00 9 3.60

62.16 9 1.66
70.83 91.49
53.16 91.19b,c
66.50 93.60

66.83 9 1.54
72.50 93.20
52.00 91.39b,c
69.00 94.70

Values are expressed as mean 9S.E.M (n = 6).

Statistically significant difference to the vehicle.
c
Statistically significant difference to the corresponding zero time value.
a

Table 3
Effect of oral administration of different doses of P. granatum extract and tolbutamide on the mean blood glucose level in alloxan
diabetic ratsa
Sampling time
(h)

0
1
2

Blood glucose level (mg/dl)

Vehicle (10% v/v Tween 80, 10
ml/kg)

Tolbutamide,
500 mg/kg

P. granatum,
300 mg/kg

P. granatum, 400
mg/kg

P. granatum, 500
mg/kg

282.83 9 4.12
277.40 9 6.50
271.839 2.15

267.169 6.60
255.009 4.95
240.009 5.65

271.66 9 4.07
193.66 9 3.96b,c
205.50 92.92b,c

280.40 9 7.80
193.16 92.15b,c
200.00 93.04b,c

274.83 92.82
189.20 93.42b,c
198.83 94.15b,c

Values are expressed as mean 9S.E.M. (n= 6).

Statistically significant difference to the vehicle.
c
Statistically significant difference to the corresponding zero time value.
a

3.3. Effect of the test drug on blood glucose

le6els in normal fasted rats
Table 2 shows the effects of various doses of
the extract of P. granatum on blood glucose level
in normal fasted rats. All doses of the test drug
extract showed maximum hypoglycaemic effect at
2 h. Tolbutamide treatment exerted a marked
time dependent hypoglycaemic effect.

3.4. Effect of the test drug on alloxan induced

diabetes in rats
Oral administration of all the doses of the test
drug extract led to significant lowering effect in
alloxan-induced diabetic rats. The fall was seen at

1 h and remained up to 2 h after administration

of the test drug extract whereas the fall in glucose
level in the case of tolbutamide administration
was marginal because alloxan causes diabetes by
destruction of the pancreatic cells and tolbutamide requires more than 30% functional pancreas for effectiveness (Table 3).

4. Discussion
This study reports for the first time, the antihyperglycaemic effect of the extract of P. granatum
(flowers), a Unani drug frequently used by Unani
physicians for the treatment of diabetes.

M.A. Jafri et al. / Journal of Ethnopharmacology 70 (2000) 309314

A clinically used tolbutamide (a sulphonylurea

drug) is known to lower the blood glucose level by
stimulating b-cells to release insulin. Since alloxan
induces diabetes by destroying b-cells and impairing renal function, in the present study tolbutamide exhibited mild hypoglycaemic activity in
the alloxan diabetic rats. However, in these rats
the extract of the flowers of P. granatum showed
marked hypoglycaemic effect. This may not be
due to potentiation of insulin release from pancreatic cells and thus the drug may also be effective
in non insulin-dependent diabetes.
Since the blood glucose lowering effect of the
extract of the flowers of P. granatum was observed in alloxan diabetic rats as well as in fasted
normal rats, this effect could, possibly be due to
increased peripheral glucose utilization. Inhibition
of the proximal tubular reabsorption mechanism
for glucose in the kidney, if any, can also contribute towards blood lowering effect (Sharma et
al., 1983).
The test drug extract also improved oral glucose tolerance in normal rats. At the present
juncture it is not possible to pinpoint the mechanism of anti-hyperglycaemic action of the extract
of P. granatum. However, based on an earlier
report some suggestions can be made for its possible mechanism. It is reported that an infusion of
the epicarps of P. granatum inhibited the intestinal absorption of glucose in rats (Nogueira and
Pereira, 1986b). Thus a possibility exists that retardation of intestinal glucose absorption may
also be partly responsible for inhibition of hyperglycaemia in glucose-fed rats.
The rats treated with 400 and 500 mg/kg of the
test drug extract showed almost the same blood
glucose lowering effect. The higher dose, i.e. 500
mg/kg, did not further lower blood glucose levels
in normal, glucose-fed hyperglycaemic and alloxan induced diabetic rats as compared to the
rats treated with 400 mg/kg. At present we may
conclude that the extract of P. granatum, in totality, was effective in reducing the blood glucose
level under our experiment conditions.
Studies are underway to further elucidate the
mechanism of hypoglycaemic effect of the extract
of the flowers of P. granatum.

313

Acknowledgements
The authors are thankful to the Department of
I.S.M. and H. (Ministry of Health and Family
Welfare) for providing financial support to our
department (Ilmul Advia) and a fellowship to one
of the authors (M.A.).

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