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Antimalarial, antiemetic and antidiabetic potential of Grewia aslatica L. leaves

Article · April 2012

DOI: 10.5897/JMPR12.028

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Journal of Medicinal Plants Research Vol. 6(16), pp. 3213-3216, 30 April, 2012
Available online at http://www.academicjournals.org/JMPR
DOI: 10.5897/JMPR12.028
ISSN 1996-0875 ©2012 Academic Journals

Full Length Research Paper

Antimalarial, antiemetic and antidiabetic potential of

Grewia aslatica L. leaves
M. Zia-Ul-Haq1*, Shakir Ahmad Shahid2, Shafi Muhammed3, Mughal Qayum4, Inamullah Khan5
and Shakeel Ahmad6
1
Research Institute of Pharmaceutical Sciences, Department of Pharmacognosy, University of Karachi,
Karachi-75270, Pakistan.
2
Department of Chemistry, University of Sargodha, Sargodha-40100, Pakistan.
3
Department of Pharmacy, University of Balochistan, Quetta, Pakistan.
4
Department of Pharmacy, Abdul Wali Khan University, Mardan 23200, Pakistan.
5
Department of Pharmacy, University of Peshawar, Peshawar-25120, Pakistan.
6
Department of Agronomy, Bahauddin Zakariya University Multan-60800, Pakistan.
Accepted 14 February, 2012

Ethnopharmacological knowledge has firm traditional and conceptual base built on a tremendous
observational data and theoretical structure but lack experimental base, and scientific evidence is
needed to prove rationale of using this indigenous flora to preserve and prosper the cultural legacy.
Researchers are blending traditional knowledge with experimental methodology for testing efficacy and
safety of these herbal remedies. The current study has been designed to authenticate antimalarial,
antiemetic and antidiabetic activities of methanolic extracts of Grewia asiatica L. leaves. The extract
revealed significant antimalarial, antiemetic and antidiabetic activity. These results indicated the
potential of G. asiatica leaves to further explore as natural source for new lead compounds against
malaria, diabetes and emesis.

Key words: Antimalarial, antiemetic, antidiabetic, Grewia asiatica L. leaves.

INTRODUCTION

Grewia asiatica L. locally known as phalsa and dhaman
is a large bushy shrub or small tree, and has height of 4
m or more. It is found throughout Punjab province of
Pakistan and has medicinal as well as food uses. Its fruits
are eaten fresh as dessert, are made into syrup, and
extensively employed in the manufacture of soft drinks.
The fruit is used in making jams, pies, squashes,
chutneys etc. However, phalsa fruit has a short shelf life
and is considered suitable only for local marketing
(Anand, 1960; Panda, 2002). The fresh leaves are valued
as fodder and are used by indigenous communities as
antimalarial aid. All parts of plant are believed to possess
anti-emetic properties. Its fruit is mordant, stomachic,
laxative, and an aphrodisiac. Unripe fruits are used for
treating fever, diarrhea, cardiac, respiratory and blood
disorders, and relieves from inflammation. The fruits
*Corresponding author. E-mail: ahirzia@gmail.com.
assuage thirst and burning sensation, and help to remove
dead foetus. Fruit extract possesses radioprotective
potential. The leaves are applied on skin rashes and
pustular eruptions due to their antibiotic potential, while
the root is used for treating strangury, pus and clap
(Morton, 1987; Pankhaj, 2004; Sharma and Sisoda,
2009; Ahmad, 2007). The plant is believed to possess
antipyretic, antidiabetic, analgesic, antifertility, antibiotic
and antimicrobial properties (Tripathi et al., 2010). The
bark is used to treat rheumatism, while infusion of bark is
used as demulcent, febrifuge and anti-diarrhoea. The
bark also cures urinary problems and alleviates burning
in vagina (Sisoda and Singh, 2009; Muhammad et al.,
2006). Besides this, baskets are made from its shoots
which are used to carry fruit and vegetables. Small tough
articles like spades; golf sticks shafts, shoulder poles for
carrying small loads, pestles, bows, billiards cues and
shingles are made from its wood. The mucilaginous
extract of the bark obtained after pounding in water is
used to purify sugar cane juice during the preparation of
3214 J. Med. Plants Res.
Table 1. Enoyl-ACP reductase inhibitory
(antimalaria) potential of Grewia asiatica.
Sample % Inhibition
G. asiatica 69.12 ± 0.01
Standard 93.17 ± 0.02
‘gur’ or brown sugar. In Burma, the bark is used as a
soap substitute while fiber extracted from the bark is
made into rope (Sosef et al., 1998). Despite its so much
medicinal and nutritional importance, very little work is
reported on this plant. In this regard as part of our
continuous studies on exploring the hidden potential of
the indigenous flora of Pakistan (Nisar et al., 2010a, b, c,
2011; Zia-Ul-Haq et al., 2007a, b, 2008a, b, 2009, 2010a,
b, 2011a, b, c, d, e) and evaluated anti-emetic,
antidiabetic and antimalarial potential of methanolic
extracts of G. asiatica L. leaves.
MATERIALS AND METHODS
Plant material and preparation of crude extract
G. asiatica L. leaves were obtained from Department of Agronomy,
Bahauddin Zakariya University Multan, Pakistan and authenticated
by Dr. Shakeel Ahmad, Assistant Professor of same department.
Plant material was cleaned and soaked in methanol for 15 days
with occasional shaking. It was filtered through a muslin cloth and
then through a filter paper. The extracts so obtained were
combined, filtered through filter paper under vacuum and
concentrated under reduced pressure in a rotary evaporator (model
Q-344B – Quimis, Brazil) using a warm water bath (model Q-214M2
- Quimis, Brazil) to obtain a thick gummy mass, which was further
dried in a desiccator and stored in air-tight vial till further use.
Antimalarial assay
Enoyl-ACP reductase was used as described earlier (Surolia and
Surolia, 2001) utilizing crotonyl CoA as a substrate and measuring
the decrease in absorbance at 340 nm using a UV-VIS
spectrophotometer at 25°C in sodium phosphate buffer (50 mM; pH
7.5) containing NaCl (100 mM). 100 µl assay solution was prepared
(Crotonyl CoA 150 µM, NADH 100 µM; PfENR 170 nM; 5% DMSO),
that served as the positive control The sample was pipetted in 96-
well UV-plates, then a mixture containing (buffer + Enzyme +
NADH) was added. This mixture was preincubated for 5 min before
initiating the reaction by adding the substrate, Crotonyl CoA. The
rate of decrease in the amount of NADH in each reaction well was
converted to percentage of inhibition using SOFTmax PRO software
(Molecular Devices, Sunnyvale, Calif.) and following formula:
Rate in the presence of sample
Inhibition (%)  100
Rate of positive control
The results are given in Table 1.
Antiemetic assay
Antiemetic assay was performed as reported previously (Tijani et
al., 2008). Four day old male chicks were divided into four groups of
five chicks each and each chick was kept in a large beaker at 25°C
for 20 min to stabilize. Group 1 served as the control and was
treated with normal saline (5 ml/kg). Groups 2 and 3 were treated
with extract (50 and 100 mg/kg) orally, while group 4 was treated
with metoclopramide (50 mg/kg) intraperitoneally. After one hour,
anhydrous copper sulphate (50 mg/kg) was administered orally to
each chick and then number of retches (an emetic action without
vomiting gastric material) was counted for 10 min. The antiemetic
effect was assessed as the decrease in number of retches in the
treated group. The inhibition (%) was calculated as follows:
Inhibition (%) =
Where A is the control frequency of retching and B is the frequency
of retching of the treated group. The results are given in Table 2.
α-Amylase inhibitory activity
The α-amylase inhibitory activity was determined by a slightly
modified method proposed by Bernfeld (Bernfeld, 1955). α-amylase
(20 µl; 0.05 U/μl) was premixed with sample (20 μl) and starch
solution (250 μl; 2%) in sodium phosphate buffer (0.1 M; pH 6.9)
was added as a substrate to start the reaction. The reaction was
carried out at 37°C for 10 min and terminated by the addition of 200
μl of DNS reagent (1% 3,5-dinitrosalicylic acid; 12% sodium
potassium tartrate in 0.4 M NaOH). The reaction mixture was
heated for 15 min at 100°C, and then diluted with 5 ml of distilled
water. α-Amylase activity was determined by measuring
absorbance at 540 nm using the following equation:
( AI  AIB )
Inhibition %  1   100
( AO  AOB )
Ai is the A540 of sample reactive solution, A0 is the A540 of control
reactive solution, AiB is the blank of sample and A0B is the blank of
control. The results are given in Figure 1.
α-Glucosidase inhibitory assay
α-glucosidase was assessed according to a previously reported
method (Matsui et al., 1996) with minor modifications. p-nitrophenyl
α-D-lucopyranoside (1 ml; 3 mM) in sodium phosphate buffer (0.2
M; pH 6.8) was added as a substrate to the mixture of α-
glucosidase (50 μl; 0.15U/µl) and sample (50 μl) to start the
reaction. The reaction was conducted at 37°C for 15 min and
stopped by the addition of Na2CO3 (750 μl; 0.1 M). The α-
glucosidase activity was assessed by measuring the release of p-
nitrophenol from pNPG at 410 nm. Glucosidase activity inhibition
was determined as percentage of control, as follows:
Where, Ai is absorbance of sample solution, and Ao is absorbance
of control solution. Acarbose was used as a positive control. The
results are given in Figure 1.
RESULTS AND DISCUSSION
Chemical analyses and biological and pharmacological

Table 2. Antiemetic activities of Grewia asiatica.
Drug/dose
Control
Metoclopramide (50 mg/kg)
G. asiatica (50 mg/kg)
G. asiatica (100 mg/kg)
100
80
Inhibition %
60
40
20
0
alpha-glucosidase
100 µg/ml 250 µg/ml
Figure 1. α-glucosidase and α-amylase inhibitory (%) activities of Grewia asiatica.
assays provide objective evidences to validate traditional
uses by indigenous communities. Research in drug
discovery involves multifaceted approach combining
botanical, phytochemical, biological and molecular
techniques. So, current study is designed to screen a
locally used plant for various tagged pharmacological
bioactivities.
Malaria is a major cause of childhood and adult illness
around the globe. Resistance to the most common
antimalarial drugs has spread to almost every part of the
world. This prompted the urgent development of new
compounds to treat malaria. Efforts are now directed
towards obtaining drugs with different structural features,
along with new strategies in malaria control and the
recognition and validation of traditional medical practices
(VanDyk et al., 2009). Our results indicate that G. asiatica
L. leaves are a potential source of antimalarial drugs. The
crude extract had maximum inhibition, that is, 69%
inhibition (Table 1). The results are very promising and G.
Zia-Ul-Haq et al. 3215
No. of retches % inhibition
50.04 ± 2.31 -
a
8.09b ± 1.65 83.9
a
30.45a ± 1.11 39.14
a
20.17b ± 2.76 59.69
alpha-amylase
500 µg/ml Acarbose 0.1 µg/ml
asiatica L. leaves should be screened to identify the
bioactive molecule that is responsible for such a higher
antimalarial activity. Plant extracts have long been used
for the ethnomedicinal treatment of diabetes in various
systems of medicine and are accepted as an alternative
for diabetic therapy because natural α-glucosidase
inhibitors from plant sources offer an attractive strategy
for the control of postprandial hyperglycemia
(Subramanian et al., 2008). The results (Figure 1)
indicated that G. asiatica L. leaves exhibited antidiabetic
potential and this potential is dose dependent, that is,
higher antidiabetic potential at higher concentrations and
lower antidiabetic potential at low concentrations.
Emesis and puking may be sign of many conditions like
gestation, peptic ulcer, drug noxiousness, renal failure,
and hepatitis, which if uncontrolled, may disturb the
quality of life by dehydration, intense metabolic
imbalances and nutrient diminution. Retching may occur
after administration of cancer chemotherapeutic agents
3216 J. Med. Plants Res.
which may lead to rejection of curative anti-neoplastic
therapy. G. asiatica leaves extract decreased significantly
vomiting frequency. Metoclopramide, which provokes
antiemetic activity by acceleration of GIT movement
(Akita et al., 1998), however was more effective than
extract. This antiemetic effect of extract may in part be
due to its effect on the visceral afferent sympathetic fibre
in the GIT and probably through agonistic effect at the
inhibitory receptor sites such as dopamine and GABA in
central nervous system (Tijani et al., 2008). Previously
antiemetic effect of G. asiatica fruit extract has been
observed on dog models (Yaqeen et al., 2008).
Development of industries based on medicinal plants
should be included as a priority area for translating lab
findings to industrial scale to break confinement from
academic pursuits. There is dire need to build up more
and better organized inter-disciplinary studies among
farmers, agricultural scientists, scientific research
institutes and herbal industries of Pakistan to add value
to medicinal plants from cultivation to marketing by
processing, chemical analysis and bioactivity-guided
fractionation to rationalize their folk uses. There is need
to conserve medicinal germplasm and catalogue of all
genetic material indigenous to Pakistan, and should be
created and maintained to hold effective intellectual
property rights (IPRs).
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