Immunomodulatory activity of Argania spinosa seeds

Abstract

Objective: Various compounds of medicinal plants have been widely investigated since ancient times for their possible immunomodulatory properties in the body's immune system. Methods: In the present study, the immunostimulatory effect of Argania spinosa crude extract was evaluated in vivo. The immunostimulant potential of crude extract on the phagocytic activity was measured by the carbon clearance rate test. Results: Our research revealed that at different doses (30,50, 100 and 150 mg/kg), Argania spinosa crude extract increased the phagocytic activity in a dose dependant manner when compared with the control and thus the clearance rate of carbon was faster after the administration of the plant extract P=0,000. Conclusion: Argania spinosa crude extract exhibited a dose-dependent immunostimulant effect on the reticuloendothelial system, which could be attributed to the presence of active principles in this crude extract.

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Research Article
IMMUNOMODULATORY ACTIVITY OF ARGANIA SPINOSA SEEDS
BOUTHEYNA ARIBI
1
, SAKINA ZERIZER*
1
, ZAHIA KABOUCHE
2
¹Department of Animal Biology, University of Constantine 1, Algeria Option Immuno-Oncology,
1*
Department of Animal Biology,
University of Constantine 1, Algeria
2
Laboratoire d’obtention de substances thérapeutiques (Lost), Université de- Constantine 1,
Laboratoire d’Obtention de Substances Thérapeutiques (L.O.S.T), Campus Chaabat Ersas, 25000 Constantine, Algeria.
Email: zerizersakina@yahoo.fr
Received: 03 Apr 2013, Revised and Accepted: 19 May 2013
ABSTRACT
Objective: Various compounds of medicinal plants have been widely investigated since ancient times for their possible immunomodulatory
properties in the body’s immune system.
Methods: In the present study, the immunostimulatory effect of Argania spinosa crude extract was evaluated in vivo. The immunostimulant potential
of crude extract on the phagocytic activity was measured by the carbon clearance rate test.
Results: Our research revealed that at different doses (30,50, 100 and 150 mg/kg), Argania spinosa crude extract increased the phagocytic activity
in a dose dependant manner when compared with the control and thus the clearance rate of carbon was faster after the administration of the plant
extract P=0,000.
Conclusion: Argania spinosa crude extract exhibited a dose-dependent immunostimulant effect on the reticuloendothelial system, which could be
attributed to the presence of active principles in this crude extract.
Keywords: Immunomodulatory, Argania spinosa seeds, Phagocytic activity, Carbon Clearance rate, Reticuloendothelial system.
INTRODUCTION
The immune system is the most complex biological systems in the
body. At the time of infection immune system go under the attack of
a large number of viruses, bacteria and fungi[1]. There are two
branches of immunity response: humoral immunity and cellular
immunity [2].
Immunity disorders may affect both cellular and humoral
components. An important role in the cellular immunity is played by
reticuloendothelial system which mainly comprise of phagocytic
cells whose function is to ensure elimination of senescent cells,
pathogenic microorganisms and immune complex from blood and
tissues and participate in inflammation. This way they contribute to
non-specific immunity. These cells also participate in specific
immunity by way of antigen presentation and cytokine secretions
[3].
In order to perform phagocytic function, cells of reticuloendothelial
system must be transformed to the active state. This specific ability
is significantly suppressed by the action of physiological and
pathological factors in nature. However, it is possible to influence
this ability using certain immunomodulating agents [4, 5]
Immunomodulation is a procedure which can alter the immune
system of an organism by interfering with its functions [6]. In the
innate immune the nature killer cell plays an important role to the
defiance against virus – infected and malignant cell to destroy the
abnormal cell [7]
Medicinal plants which are used as immunomodulatory effect to
provide alternative potential to conventional chemotherapy for a
variety of diseases, especially in relation to host defense mechanism.
The use of plant product like polysaccharides, lectines, peptides,
flavonoids and tannins has been the immune response or immune
system in various in –vitro modals [8].
Argan tree (Argania spinosa (L.) Skeels) belongs to the Sapotaceae
family and it is the only species of this tropical family, is endemic in
South-western Morocco [9] and Algerian region of Tindouf [10, 11].
The fruit of the argan tree is a stone-fruit (as for walnut tree or
almond tree); with pulp covering a lignified endocarp (the nut)
containing one to three kernels (the seeds) which furnish edible and
marketable oil [12].
Pharmacological studies have confirmed that Argania spinosa have
several biological effects including: antiproliferative [13, 14, 15, 16]
Hypolipidemic, hypocholesterolemic [17], antiatherogenic [18, 19,
20] antiradical [21, 22] and anti-inflammatory activities [23].
The present investigation was undertaken to evaluate the
immunostimulatory effect of the crude extract obtained from
Argania spinosa seeds using phagocytic responses by carbon
clearance test in vivo experimental model.
MATERIALS AND METHODS
Plant Material
Seeds of Argania spinosa were collected from Tindouf (South –west
of Algeria).
The fruits were cut into pieces to obtain seeds, and then the seeds
were subjected to size reduction to a coarse powder using a
mechanical grinder. The powder (crude extract) was then used for
treatment preparations by dissolving it in normal saline (0.9%).
Animals
Adult male Mus Musculus mice ( 2.5- 3 month old ) from central
pharmacy Algeria, weighing(28- 35) were used for determination of
the phagocytic activity.
The animals were kept under standard laboratory conditions of
humidity, temperature (25± 1°C) and light (12h day :12h night), and
allowed free access to food and water. The animal studies were
conducted after obtaining clearance from Institutional Animal Ethics
Committee and the experiments were conducted in strict compliance
according to ethical principles and provided by Committee for the
Purpose of Control and Supervision of Experiments on Animal(
CPCSEA).
Phagocytic index
The clearance rate of carbon was measured by the method of [24].
Animals were divided into five groups, consisting of six mice in GI,
GII, GIII, GIV and GV. Group I (control) was given 0,9% Nacl (0,5
ml/mouse i.p.), Groups II-III-VI and V were administered by i.p
injection with different concentrations of Argania spinosa crude
extract (30, 50, 100 and 150 mg/kg/) respectively.
International Journal of Pharmacy and Pharmaceutical Sciences
ISSN- 0975-1491 Vol 5, Issue 3, 2013
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After 48h of i.p injection, the mice were administered with carbon
ink suspension at a dose of ( 0.1ml/10g through the tail vein ; the
mixture consisted of black carbon ink 3ml, saline 4ml and 3%
gelatine solution 4ml.
Blood samples were taken from the retro orbital vein by using glass
capillaries, at 5 and 15 min. Blood sample drops (14) were mixed
with 0.1% sodium carbonate solution (4ml) for the lysis of
erythrocytes and the absorbance measured at 675 nm using a
spectrophotometer.
The animals were sacrificed and the liver and spleen dissected and
weighed immediately in the wet state.
The phagocytic activity is expressed by the phagocytic index K which
measures all the reticuloendothelial system function in the contact
with the circulating blood and by corrected phagocytic index α
which expresses this activity by unit of active weight organs: liver
and spleen. The clearance rate is expressed as the half-life period of
the carbon in the blood (t
1/2
, min). These are calculated by means of
the following equations [25, 26]:
Where OD
1
and OD
2
are the optical densities at times t
1
and t
2
respectively.
Statistical Analysis
Results were analysed for differences between the groups across
dietary treatments by one –way ANOVA test and Tukey’s multiple
comparison tests (SPSS version 9).
RESULTS
The present data showed that there is a highly significantly
difference in the means for the phagocytic index (K) between groups
(GI, GII, GIII, GIV and GV) P= 0,000 and the group V is Highly
significantly different from groups ( GI,GII,GIII and GIV) at P=0,000.
This indicates that Argania spinosa crude extract enhanced the
phagocytic activity by stimulating the reticuloendothelial system in a
dose dependant manner (Figure 1).
Fig. 1: Effect of Argania spinosa crude extract on phagocytic activity.
As shown in Figure 2, the half-time of colloidal carbon was highly
significantly faster at 48h, after the administration of Argania
spinosa crude extract between groups P= 0,000 and the clearance
rate was decreased highly and significantly in groups (GII,GIII,GIV
and V) when it is compared to the control group(GI) P=0,000. This
indicates that the extract reduces the elimination time of carbon
particles from blood and affirms that Argania spinosa crude extract
enhanced the phagocytic activity.
Fig. 2: Effect of Argania spinosa crude extract on half time t1/2 of carbon in blood.
The results of this study showed that there is a highly significantly difference in the means for the corrected α between groups (GI, GII,GIII, GIV and
GV) P= 0,000 and the corrected α was increased highly and significantly in groups ( GII,GIII,GIV and V) when it is compared to the control group
(GI)P=0,000. Figure 3

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Int J Pharm Pharm Sci, Vol 5, Issue 3, 488-491
490
Fig. 3: Effect of crude extract of Argania spinosa seeds on corrected phagocytic index
DISCUSSSION
Due to high cost of antiretroviral drugs have caused researchers to
turn to plants as prospective therapies in the search of alternative
anti HIV or immunomodulatory compounds [27].
There are a number of natural agents (herbs) which are used for the
enhancing of the body’s response to disease. In recent time a large
number of drugs extracted from the plants are coming in to the
marked by proper clinical trials. When taking any of these agents
take proper advice on dose, length of treatment [1].
In this study we observed that the animals administered with the
crude extract of Argania spinosa stimulates the phagocytic index at
different concentration. So, this result agrees with those of Shuklaa
et al. [28] and Benmebarek et al. [29] who reported that the oral
administration of ethanolic extraction of Caesalpinia bonducella and
S. mialhesi respectively in the mouse are increased the phagocytic
index at different concentration.
Also Gaoa et al. Have reported that the extract of Curcumin from
Curcuma longa is inhibited the IL-2 induced proliferation of spleen
cells completely at concentrations of 25mmol/L [30]. The study of
[31] confirmed that the methanolic extract of S. mahagoni seeds has
therapeutic potential and could be served as an effective
immunomodulatory candidate without any side effects
Treatment by Argania spinosa crude extract enhanced the rate of
carbon clearance from the blood when it is compared to the control
group. This reflects the enhancement of the phagocytic activity of
phagocytes and non specific immunity, which includes opsonisation
of the foreign particulate matter with antibodies and complement
C3b, leading to a more rapid clearance of foreign particulate matter
from the blood [32].
[33] Showed the immunomodulatory activity of Isoprinosine on
chicken infected by three different viruses : Newcastle disease, fowl
plague and avian infectious bronchitis. A stimulatory influence on
primary anti-Newcastle disease virus antibody response was
observed. In the avian model the Isoprinosine antiviral effect
appears as due mainly to the enhancement of interferon production
and to a synergistic interferon –isoprinosine interaction.
CONCLUSION
In vivo investigations showed that the crude extract of Argania
spinosa at concentrations of 150mg/kg increased the phgocytic
index, corrected α and decreased the rate of carbon clearance this
immunomodulatory effect of Argania spinosa could be attributed to
its interesting chemical composition. It is essentially characterized
by the presence of unsaturated fatty acids, antioxidant compounds
(Vitamin E family), phenolic compounds, triterpenoids, sterols and
saponins [34, 35].
ACKNOWLEDGEMENT
The authors are grateful to the DG-RSDT at the MESRS (Ministry of
Scientific Research, Algeria) for the financial support. I thanks Mr
Racim Boudjemili for grammar corrections.
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