Leaflets 13: 47-50. 2009.
Investigations on Hepatoprotective
Activity of Leaf Extracts of Aegle
marmelos (L.) Corr. (Rutaceae)
T. Kalaivani1, N. Premkumar1, S.Ramya1, R. Siva1,
V. Vijayakumar2, E. Meignanam1,
C.Rajasekaran1* and R. Jayakumararaj3
of Biotechnology, Chemical and Biomedical Engineering, VIT
– 632 014.
of Humanities and Social Sciences, VIT
– 632 014.
Department of Botany, RD Govt. Arts
College, Sivagangai – 630561
Author: Dr. C. Rajasekaran, School
of Biotechnology, Chemical and
Biomedical Engineering, VIT University,
Vellore – 632 014
Issued 04 January 2009
present study was carried out to screen and evaluate the hepatoprotective activity of leaf extracts of Aegle
marmelos (L.) Corr. Hepatoprotective
activities of ethanolic and aqueous extracts of A. marmelos were examined against carbon tetrachloride induced
liver damage in mice using silymarin as control. Enzyme activities of Serum
Glutamate Oxaloacetate Transaminase (SGOT), Serum Glutamate Pyruvate
Transaminase (SGPT) and Alkaline Phosphatase (ALP) were analyzed. Results
indicate that ethanolic and aqueous leaf extracts of A. marmelos had moderate activity over carbon tetrachloride treatment
as compared to control. Results of the present investigation confirm the traditional
uses of this plant as a potential hepatoprotective agent.
KEYWORDS: Aegle marmelos; Hepatoprotective activity; Serum Glutamate Oxaloacetate Transaminase
(SGOT); Serum Glutamate Pyruvate Transaminase (SGPT); Alkaline Phosphatase
plants form the backbone of traditional system of medicine in India. Pharmacological studies have acknowledged the value of
medicinal plants as potential source of bioactive compounds (Prusti et al.,
2008). Phytochemicals from medicinal plants serve as lead compounds in drug
discovery and design. Medicinal plants are rich source of novel drugs that
forms the ingredients in traditional systems of medicine, modern medicines,
nutraceuticals, food supplements, folk medicines, pharmaceutical
intermediates, bioactive principles and lead compounds in synthetic drugs
(Ncube, 2008). WHO pointed out that more than 80% of world’s population
depends on plants to meet their primary health care needs. However,
overexploitation of the selected medicinal plant species lead to the
reduction in number of plants in the wild and inclusion of their name in the
red data book (Ahmedullah and Nayar 1999).
Aegle marmelos (L.)
Corr., belongs to the family Rutaceae, and is popularly known as Bael
tree (Eng.) (Gamble,
1935; Mathew, 1983). In Hindu mythology leaves and wood of Aegel marmelos are used to worship
Lord Shiva. This is a sacred tree amongst the Hindus. This tree is commonly found
in Hindu scared grooves. It is considered sacrilegious to destroy it;
enormous quantities of the leaves are gathered for use during ritual
ceremonies. In ancient time it is frequently alluded to as an emblem of
fertility (Jain and Sastry, 1979). Hindu physicians regard the unripe or half
ripe fruit as astringent, digestive, and stomachic, and prescribe it for diarrhoea
and dysentery (Bakhru, 1997). The ripe fruit is aromatic, cooling and is used as laxative.
The root bark is used as a remedy in hypochondriasis, melancholia and
palpitation of the heart. The fresh juice of the leaves is taken with honey
as a laxative and febrifuge; it is used in asthmatic complaints. Addition of
black pepper in anasarca is used to treat costiveness and jaundice; moreover,
in external inflammations it is given to correct the supposed derangement of
the humours. Small unripe fruit is consumed with fennel seeds and ginger in
decoction for piles (Kamalakkannan and Prince, 2005). The fruit
is used as a remedy for diarrhoea. Two tols of bark juice is given with
a cummin in milk to increase the quality of seminal fluid.
The tribals in Salem,
Dharmapuri, Vellore regions, Tamilnadu,
India offer leaves in
the month of July/ August, to god to overcome sterility problem and
subsequent year the couples are blessed with the child. Therefore, this tree is considered as an emblem
of fertility. Beverages prepared with fruit pulp are used to relive body
heat. Cologne is obtained by
distillation from flowers. The wood is used for carving, small-scale turnery,
tool and knife handles, pestles and combs, taking a fine polish. The ripen
fruit, tamarind and sugar in mixture is used as laxative to overcome
constipation and body heating problems (Jain and Sastry, 1979). Mature but
still unripe fruits are made into jam. A firm jelly is made from the pulp
alone or better still, combined with guava to modify the astringent flavor.
The pulp is also pickled (Bakhru, 1997). The shell of hard fruits has been
fashioned into pill- and snuff boxes, sometimes decorated with gold and
silver. The gum from seeds is used as household glue and as an adhesive by
jewelers. It is used to wash the silver ornamentals and shields. The fruit pulp is used as detergent for washing
clothes. The fruit pulp is used as shampoo. The dried pulp is also used in
local based hair cosmetics along with mehandi and amla. In the present study we have evaluated the hepatoprotective
potential of A. marmelos.
MATERIALS AND METHODS
leaves of A. marmelos were collected from Vellore, Tamilnadu, India during Apr 2008. The Flora of Presidency of Madras (Gamble, 1935) and The Flora of Tamil Nadu Carnatic
(Matthew, 1983) were used for identification and authentication of the
plants. Collected material was washed thoroughly in running tap water, rinsed
in distilled water and shade dried in open air and grounded into powder.
leaves were dried under shade and coarsely powdered. The powder was
successively extracted using soxhlet apparatus with ethanol and water. These
extracts were condensed using rotary vacuum evaporator followed by vacuum
evaporator and stored in desiccator. The powder of all the extracts was
suspended in appropriate solvent systems and was subjected to further
The cross breed albino mice
weighing 20-25g were housed in clean propylene cages and maintained at 30±2oC
under natural light/ dark conditions. They were fed with standard pellet diet
and water was given ad libitum. The animals were acclimatized
to laboratory conditions for 2 weeks. Animals were divided into seven groups
of six mice each. The body weight of each of the animals was recorded
initially. The period of experimentation was 5 days.
tetrachloride induced hepatotoxicity
Group I animals received 1% CMC
in distilled water (2ml/ kg body weight) for five days with olive oil (2ml/kg
body weight i.p) on second and third day. Group II animals received 1% CMC
(2ml/kg body weight) for 5 days with 1: 1 mixture of olive oil and CCl4
(2ml/kg body weight i.p.) on 2nd and 3rd day. Group III
animals served as positive control and were given silymarin (200mg/kg) for
five days. Group IV and Group V animals were given ethanolic extract of AM
(500mg/kg) and (600mg/kg) body weight respectively. Group VI and Group VII
animals received respectively 500mg/kg and 600mg/kg body aqueous extract of
AM weight. In addition to ethanolic and aqueous extracts, 1:1 mixture of
olive oil and CCl4 (2ml/kg body weight i.p.) were given to Group
IV to Group VII animals on 2nd
and 3rd day.
On the fifth
day, blood was collected from the retro orbital plexus of the animals and
serum was allowed to coagulate at 370C for 30 mins, and subjected
to centrifugation at 2500 rpm. Serum samples were stored at 2-80C
until further use. The enzyme assay was determined for Serum Glutamate
Oxaloacetate Transaminase (SGOT), Serum Glutamate Pyruvate Transaminase (SGPT)
(Reitman, 1957), and Alkaline Phosphatase (ALP) (modified method of Kind, 1954)
using commercially available enzyme kit (Crest Biosystems, Goa,
RESULTS AND DISCUSSION
Levels in CCl4 induced hepatotoxicity
with respect to SGOT, SGPT and ALP in mice using aqueous and ethanolic
extracts of A. marmelos is given in
Table 1. As compared to the control (61.3 ± 5.2, 41.3 ± 2.9, 5.7 ± 0.4),
CCl4 treated animals exhibited significantly higher levels of
enzyme activities (142 ± 11.5a, 100 ± 8.7a, 12.0 ± 0.2a)
in serum. Ethanolic extract (500mg/kg) (99.3 ± 7.8b, 53.3 ± 2.9b,
6.0 ± 0.3a) was found to have moderate activity as compared to
silymarin 54 ± 5.7a, 35.3 ± 1.7a and 4.4 ± 0.3a for
SGOT, SGPT and ALP respectively. Analysis of SGOT, SGPT and ALP levels in carbon
tetrachloride induced hepatotoxicity in mice against aqueous and ethanolic
extracts of A. marmelos revealed
that ethanolic and aqueous extracts were moderately effective when compared
to silymarin treatment. Further, the P<0.01 values in the case of AmEE (500
mg/kg) was significant indicating that ethanolic extracts of A. marmelos holds a potential to be
used as an hepatoprotective agent.
are known to have beneficial therapeutic effects documented in Traditional
Indian System of Medicine. Much work has been done on ethnomedicinal plants
in India. Interest in a large number of traditional natural
products has increased. It has been suggested that phytochemical extracts
from A. marmelos, antidiabetic, antitumoral and antimicrobial agents
(Rajsekaran et al., 2008). Results indicate that ethanolic extracts of
A. marmelos holds a potential to be
used as hepatoprotective agent. Further, this investigation acknowledges the ethnobotanical
uses and hepatoprotective nature of A.
The authors are thankful to VIT
Management for their constant support and encouragements. Thanks are due to
Prof. Lazar Mathew for his valuable comments and suggestions to carry out
this research successfully.
Ahmedullah M and Nayar MP (1999). Red data
book of Indian plants, (Peninsular India), Calcutta:
Botanical Survey of India. Vol. 4.
Bakhru HK (1997) Foods that Heal. The Natural
Way to Good Health. Orient
Paperbacks. ND, India.
Gamble JS (1935)
Flora of the Presidency of Madras. Adlard and Son’s Ltd, London, UK.
and Sastry ARK
(1979). Threatened Plants in India.
Botanical Survey of India.
Calcutta, WB, India.
N and Prince PS (2005). The effect of Aegle marmelos fruit extract in streptozotocin diabetes: a
histopathological study. J Herb Pharmacother 5:87-96.
Matthew KM (1983)
The Flora of Tamil Nadu Carnatic. In The Rapinat Herbarium. St Joseph's College, Tiruchirapalli, India
Ncube NS, Afolayan AJ and Okoh A (2008) Assessment techniques of
antimicrobial properties of natural compounds of plant origin: current
methods and future trends. African
Journal of Biotechnology 7(12):1797-1806.
Prusti A, Mishra SR,
Sahoo S and Mishra SK (2008) Antibacterial Activity of Some Indian Medicinal
Plants. Ethnobotanical Leaflets 12:
Meignanam E, Premkumar N, Kalaivani T, Siva R, Vijayakumar V, Ramya S and
Jayakumararaj R (2008) In Vitro
Evaluation of Antibacterial Activity of Phytochemical Extracts from Leaves of
Aegle marmelos (L.) Corr.
(Rutaceae). Ethnobotanical Leaflets
12: 1124-28. 2008.
Table 1. Analysis of hepatoprotective activity of
leaf extracts of Aegle marmelos.
Control (1% CMC)
61.3 ± 5.2
41.3 ± 2.9
5.7 ± 0.4
142 ± 11.5a
100 ± 8.7a
12.0 ± 0.2a
Silymarin (200 mg/kg) + CCl4
54 ± 5.7a
35.3 ± 1.7a
4.4 ± 0.3a
AmEE (500 mg/kg) + CCl4
99.3 ± 7.8b
53.3 ± 2.9b
6.0 ± 0.3a
AmEE (600 mg/kg) + CCl4
115.3 ± 8.5c
80 ± 4.1c
9.8 ± 0.2c
AmAE (500 mg/kg) + CCl4
120.2 ± 6.1c
85.6 ± 4.7c
9.6 ± 0.4b
AmAE (600 mg/kg) + CCl4
111.3 ± 6.3c
82 ± 5.2c
9.0 ± 0.3b
AmEE (600 mg/ kg) treated
Silymarin (200 mg/ kg) treated
AmAE (500 mg/ kg) treated
AmEE (500 mg/ kg) treated
AmAE (600 mg/ kg) treated
Values are expressed in Mean ± SE for three
animals in each group; aP<0.001; bP<0.01; cP<0.1.