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International Journal of Marine Science 2013, Vol.3, No.19, 151-157
http://ijms.sophiapublisher.com
152
system to explore the bioactive potential of any
natural or synthetic drug so as to extrapolate the
results to human immune system. In order to screen
the natural products for immunomodulating activities,
a number of studies had been carried out using rodent
models. The aqueous suspension of dried latex of
Calotrophis procera
showed anti-inflammatory property
in rat models (Ranjit et al., 2008). The water extract of
Azadirachta indica
exerted significant anti-inflammatory
activity in the cotton pellet granuloma assay in rats
(Ray et al., 1996).
Gupta et al (2006) has highlighted the therapeutic
potential of immunomodulatory agents from plant
products. Galina et al (2009) had listed out several
plants that were proved to be immunomodulators
(Kannan et al.,
2009). The extracts of the plant
Rubia cardifolia
have been tested successfully for
immunomodulatory activities. (Like plant bioactive
material several immunomodulative bioactive compounds
had been isolated from marine natural products.
The bioactive compound Latrunculin A isolated from
Latrunculia magnifica
was found to inhibit prostrate
tumor cell invasion and hypoxia inducible factor-1
activation of breast tumor cells (El Sayed et al.,
2008).
The Spongistatin compound was isolated from
Spongia
sp. induced apoptosis in leukemia cells
including those that over express the protein. The
sponge
Suberea mollis
was found to contain
subereaphenol A that show antioxidant activity
(Abou-shoer et al., 2008). Acanthomine A isolated from
Acanthostrongylophora
sp. was reported to be cytotoxic
to mammalian cancer cell (Ibrahim et al., 2008).
Likewise many new compounds are isolated from
marine sponges to stimulate or suppress immunity. As
the availability of marine sponge derived bioactive
compound is meager, chemical synthesis of the
bioactive compounds are carried out to develop
possible drugs for immunomodulation. Hence an
attempt has been made to identify few bioactive
compounds in the sponge
A. globostellata
(Ag)
to
evaluate the immunity modifying activity using
clinical trials in rodent model.
2 Materials and Methods
2.1 Sponge and extract preparation
Sponge
Aurora globostrellata
(7.85 kg each) were
collected from Rameswaram (9°28′N, 79°12′E) coastal
area and washed with sea water, air dried and chopped
into small size before being ground into fine paste.
The sponge was successively extracted with solvents
with an increasing polarity (Ethyl acetate). The sponge
extracted material was used for further analysis.
2.2 Immunity study in murine system
2.2.1 Animals and treatment
For the experimental study, Wister rats were chosen.
The rats were obtained from Madras Medical College,
Chennai and reared in laboratory under standard
conditions of light and darkness (12
-
12 h) and
temperature (22±2
℃
). The rats were fed Standard
laboratory mice pellet feed (Lipton Ltd., Mumbai)
(Consisting protein 15~17 %, fat 4~5 %, carbohydrate
45~55 %, fiber 15%, vitamin A 7000 IU/kg, vitamin E
40 mg/kg, vitamin K 2 mg/kg, vitamin B l mg/kg
Hawk-Oser salt 11 mg/kg) and water
ad libitum
to all
the animals. The access to animal room was limited and
kept to minimum.
For the experimental study rats weighing 150 to 210
gm (30 days old) were recruited from the acclimatized
stock. The rats were grouped into several groups and
each group with six individuals. These animals were
housed in a specially designed (polyethylene cage)
cage with provision for systematic supply of pellets and
water. The animals were trained to take water from a
feeding bottle kept in cage.
Sponge
Aurora globostrella
extract and standard drug
treatment was given to animals for 3 to 5 weeks.
During treatment pellet feed and water were given in
ad libitum.
Food consumption, general condition and
other symptoms were observed daily and body
weights were recorded weekly. For treatment with
sponge purified compound, the doses for the treatment
were fixed three groups. In the present study the
bioactive compounds were separated using column
chromatography. The sixteenth fractions separated
from the column showed many bioactive compounds.
The dominant peak showed the presence of
3-hydroxytetradecanoic acid. From these compound
fractions three different concentrations were prepared
using Alsevier's solution
.
Three different concentrations
were 50 mg/kg, 150 mg/ kg and 250 mg/kg.
2.2.2 Immunization
The test animals were divided into equal groups for
stimulation with sheep red blood cells (SRBC). For