Ethnobotanical Leaflets 12: 1206-12. 2008.
Immunomodulatory Effect of Coconut Protein on Cyclophosphamide Induced Immune Suppressed Swiss Albino Mice
1A. Geo Vigila and 2X. Baskaran
1Department of Biochemistry,
2 Department of plant biology and plant biotechnology,
immunomodulatory properties of coconut protein were investigated in Swiss
albino mice by immune suppressed with cyclophosphamide (CP). CP is
a commonly used anti-cancer drug which causes toxicity by its reactive
metabolites such as acrolein and phosphoramide mustard. In this study, the animals were grouped
into four of 6 mice per group. Assessment of immunomodulatory activity was
carried out by testing the
Key words: Cyclophosphamide, coconut protein, immunomodulatory,
The term immunomodulatory means regulation of the immune system by suppression and stimulation of the cells and organs of the immune system (Bafuna and Mishra, 2005). Immunostimulation in a drug-induced immunosuppression model and immunosuppression in an experimental hyper-reactivity model by the same preparation can be said to be true immunomodulation (Patwardhan et al., 1990). Certain agents have been shown to possess activity to normalize or modulate pathophysiological processes and are hence called immunomodulatory agents (Wagner, 1984). A number of plant products are being investigated for immune response modifying activity (Upadhyay, 1997). Cyclophosphamide acts on both cyclic and intermitotic cells, resulting in general depletion of immune-competent cells. Cyclophosphamide (CP) is an alkylating agent widely used in anti-neoplastic therapy (Baumann and Preiss, 2001; Fleming, 1997). It is effective against a variety of cancers such as lymphoma, myeloma and chronic lymphocytic leukemia (Baumann and Preiss, 2001). CP-induced immunosuppression is reported to prompt various types of infection (Angulo et al., 2002; Wijesundera, 1980; Williams and Luzio, 1979).
Coconut palm (Cocos nucifera L.) is an
economic plant which is cultivated in most tropical countries. A coconut
fruit is composed of about 38.5% shell, 51.7% kernel and 9.8% water (Dendy
& Timmins, 1973). Hypolipidemic effect of coconut protein is due
to the high content of L-arginine (Mini et
al., 2004), feeding coconut kernel along with coconut oil in human
volunteers has been found to reduce serum total and
Separation of Coconut Protein
The nuts were freed from shells and skins and ground to a fine paste and suspended in 10% NaCl solution. Toluene was added to remove fat. The mixture was stirred at room temperature for 24 h. The solution was freed from oil and fat by filtering and centrifuging. Saturated ammonium sulfate was added to 60% saturation and the precipitate was filtered and washed with 50% saturated ammonium sulfate solution. The resulting protein precipitate was dissolved in 0.033 M sodium phosphate buffer, pH 6.7, containing 7% NaCl. The protein solution was then dialyzed against the same buffer for 6 days (Balasundaresan et al., 2002).
Preparation of feed
The animals were fed with a synthetic diet:
Coconut protein - 120 g
Sago - 315 g
Groundnut oil - 35 mL
Vitamins - 30 g
Sources of Laboratory Animals
the present study, healthy adult male albino mice were used as experimental
animals. The animals were purchased from
Before starting the experiments the animals were
acclimatized to the laboratory condition for 7 days. After acclimatization of
the laboratory condition, animals were pretreated with coconut protein for 30
days. CP (20 mg/kg/day) was given orally for 10 days for the animal of Group-II
Evaluation of the Immunomodulatory Activity of Coconut protein on Experimental Animals
Twenty four Swiss albino mice were used to assess
the effect of coconut protein on the immune system. The mice were divided
into 4 groups of six mice per cage. The first group was given normal diet and
water and this (Group-I) served as control. The second group served as test
group (Group-II) fed with normal diet and induced with the CP. The third
group served as experimental animals (Group-
At the end of the treatment, mice were sacrificed
under chloroform anesthesia. The blood was collected through cardiac puncture
and blood was collected immediately. Haematological parameters were studied
for White Blood Cell (WBC), Red Blood Cell (
count was done by Turke’s method,
The data were expressed as the mean ± standard deviation of the means (S.D) and statistical analysis was carried out employing student’s t-test.
Modification of immune functions by pharmacological
agents is emerging as a major area of therapeutics. The
Table 1: The
Bone marrow is a site of continued proliferation and turnover of blood cells, and is a source of cells involved in immune reactivity (Pelczar et al., 1990). A high degree of cell proliferation renders bone marrow a sensitive target, particularly to cytotoxic drugs. In fact, bone marrow is the organ most affected during any immunosuppression therapy with this class of drug. Loss of stem cells and inability of bone marrow to regenerate new blood cells will result in thrombocytopenia and leucopenia. The results indicate modulation of bone marrow activity, namely suppression when used CP alone and stimulation to counteract with CP-induced pretreated coconut protein groups.
Table 2: The results of differential counts of normal and pretreated groups.
and other key cells of the immune system are known to activate production of
antibody polymorphonuclear granulocyte to destroy the antigen (
Coconut protein has the ability to stimulate the immune system by activating the bone marrow cells. By activating the cells, the treated animal parameters showed similar normal condition. The increased levels of the immune cells showed a positive result. The result obtained in the present study has showed the immunomodulatory activity of coconut protein in vivo.
Cyclophosphamide is a potent suppressor of immune function, at high doses resulting in a sustained decrease in both the number and function of T and B cells (Cupps et al., 1982). It is an effective therapy in certain autoimmune diseases. Plant based immune stimulation may also contribute to the therapy of the autoimmune diseases. Coconut protein has counteractivity to cyclophosphamide-induced myelosuppression and thrombocytopenia. Thus, Coconut protein has immunostimulatory activity with regard to IgE mediated and cell-mediated hyper-reactivity. Further studies are required to examine the exact mechanism of CP function.
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