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131]

Original Article

The Potency of Nanoparticle of Pinus merkusii as

Immunostimulatory on Male Wistar Albino Rat
Sri Agus Sudjarwo, Giftania Wardani1, Koerniasari Eraiko2, Koerniasari3
Department of Pharmacology, Faculty of Veterinary Medicine, Airlangga University, 1Department of Pharmacy Biology, Faculty of Pharmacy, Hang Tuah University,
2
Department of Conservative Dentistry, Faculty of Dentistry, Airlangga University, 3Study Program of Environmental Health, Polytechnic of Health,
Surabaya, Indonesia

Abstract
Objective: Medicinal herbs are the commonly used worldwide immunomodulators in the management of various disease conditions. The aim
of this study was to evaluate the immunostimulatory activity of the nanoparticle extract of Pinus merkusii in Wistar albino rats. Materials and
Methods: It was an experimental study that was conducted on various groups of animals each with six healthy adult rats. Neutrophil adhesion
test, hemagglutinating antibody (HA) titer, delayed-type hypersensitivity (DTH) response, phagocytic activity, and cyclophosphamide-
induced myelosuppression were determined in various groups of animals. Results: The nanoparticle of extract P. merkusii at doses 500 mg/kg
BW but not at doses 125 mg/kg BW and 250 mg/kg BW induced a significant increase in percent neutrophil adhesion fibers as well as a dose-
dependent increase in antibody titer values and potentiated the DTH reaction induced by sheep red blood cells. Also, it prevented
myelosuppression in cyclophosphamide drug-treated rats and good response toward phagocytosis in carbon clearance assay. Conclusion:
From these findings, it can be concluded that nanoparticle extract of P. merkusii possesses immunostimulatory activity and has therapeutic
potential for the prevention of immune-depressed conditions.

Keywords: Immunostimulatory, nanoparticle, Pinus merkusii

INTRODUCTION immunostimulatory in traditional medicines, for the treatment

Immunomodulatory is a substance which suppresses or
stimulates the components of immune system including both
innate and adaptive the immune responses. It has been reported
that immunomodulation of the immune response could provide
an alternative to conventional chemotherapy for a variety
of disease conditions, especially when impaired immune
responsiveness of the host’s in under conditions or when a
selective immunosuppressant has to be induced in a situation
like autoimmune disorders and organ transplantation.[1,2]
The immunostimulation of the immune system of medicinal
plant products has become subject to scientific investigations
currently worldwide and their active components provide
a potential alternative to conventional immunotherapy
for a variety of immunologic diseases. In this context,
the development of medicinal plant product-based drug
candidates as immunostimulatory has gained momentum in
research studies directed toward design and discovery of
drugs.[3,4] Medicinal plant products have long been used as
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of many immunological disorders. Andrographis paniculata,
Allium sativum, Cajanus indicus, Gymnema sylvestre,
Asparagus racemosus, Piper longum Linn., Curcuma longa,
Phyllanthus emblica Linn., Ocimum sanctum Linn., Tinospora
cordifolia, and Pinus radiata are among the medicinal plants
claimed to possess potential immunostimulatory agent.[5,6]
It has been demonstrated that the medicinal properties of Pinus
plant were due to the phytochemicals possessed, including
alkaloids, polyphenols, flavonoids, lignans, triterpenes, sterols,
glycosides, triterpenoids, and saponins.[7,8] Recent research
activities have shown that Pinus plant is an important source
of pycnogenol that contains proanthocyanidins (procyanidins).
[9-11]
Proanthocyanidins are potent, free radical scavengers,
Address for correspondence: Prof. Sri Agus Sudjarwo, Department of
Pharmacology, Faculty of Veterinary Medicine, Airlangga University,
Surabaya - 60115, Indonesia.
E-mail: ags158@yahoo.com
This is an open access article distributed under the terms of the Creative Commons
Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix,
tweak, and build upon the work noncommercially, as long as the author is
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For reprints contact: reprints@medknow.com

How to cite this article: Sudjarwo SA, Wardani G, Eraiko K,

DOI: Koerniasari. The Potency of Nanoparticle of Pinus merkusii as
10.4103/ijnpnd.ijnpnd_72_17 Immunostimulatory on Male Wistar Albino Rat. Int J Nutr Pharmacol
Neurol Dis 2018;8:10-5.

10 © 2018 International Journal of Nutrition, Pharmacology, Neurological Diseases | Published by Wolters Kluwer - Medknow
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Sudjarwo, et al.: The potency of nanoparticle of Pinus merkusii as immunostimulatory
antibacterial agents, exhibit vasodilatory, antiallergic, anti-
inflammatory, cardioprotective, immune-stimulating, antiviral,
and estrogenic activities.[12,13]
In recent years, synthesis of nanoparticles is an interesting
issue of the nanoscience and nanobiotechnology. There is a
growing attention to biosynthesis the nanoparticles using
herbal. Among these organisms, plants seem to be the best
candidate, and they are suitable for large-scale biosynthesis
of nanoparticles. Nanoparticles produced by plants are
more stable, and the rate of synthesis is faster than that
in the case of other organisms. Moreover, the nanoparticles
vary greatly in shape and size in comparison with
those produced by other organisms.[14,15] Pinus plant
nanoparticles have drawn the attention of researchers
because of their suitable applications in the fields of
material science and medicine.[12] The objective of the
present study was to evaluate the immunostimulatory
activity of the nanoparticle extract of Pinus merkusii in
Wistar albino rats.
MATERIALS AND METHODS
Experimental animals
Male Wistar albino rat weighing approximately 200–250 g
(2.5–3 months) were obtained from Gadjah Mada University,
Yogyakarta, Indonesia for experimental purpose. They were
housed in plastic cages in an air-conditioned room with a
temperature maintained at 26 ± 2°C and 12 h alternates light
and dark cycles. The rats were given ad libitum with tap water
and fed with standard commercial rat chow. This study was
reviewed by the Ethical Clearance Committee for preclinical
research, Institute of Tropical Disease, Airlangga University
and obtained ethical clearance under No. 93/ITD/5/2017.
Preparation of ethanol extract of P. merkusii
Plant material and extract preparation of P. merkusii leaf were
collected from Surabaya, Indonesia. P. merkusii leaf
materials were cleaned with running tap water and
chopped into pieces. They were dried under shade at
ambient temperature for 5 days, and the air-dried
P. merkusii were then ground to powder for extraction.
The powdered P. merkusii (1 kg) was macerated with
ethanol (5 L) for a week at 37°C. The supernatant was
then collected and filtered through Whatman No. 1 filter
paper in a Buchner funnel under vacuum. The filtrate was
concentrated by evaporation with a vacuum rotary evaporator
at 45°C. The extract was dried at reduced pressure, stored at
0–4°C, and used for the experimentation.
Preparation of P. merkusii extract nanoparticle
P. merkusii extract nanoparticles were prepared by ionic
gelation of chitosan and sodium tripolyphosphate (TPP)
anions. One gram of P. merkusii extract was dissolved in
50 ml Tween 80 solutions and added 100 ml chitosan in
glacial acetic acid (0.25% v/v). Three hundred and fifty
milliliters of 0.84% w/v TPP was added under magnetic
International Journal of Nutrition, Pharmacology, Neurological Diseases ¦ Volume 8 ¦ Issue 1 ¦ January-March 2018
stirring at room temperature in drop wise using 10 ml
syringe needle into P. merkusii extract and chitosan
solution. The pH of the solution was adjusted by adding
0.1 M NaOH solution to the chitosan P. merkusii complex and
stirred for 2 h on a magnetic stirrer. Chitosan P. merkusii
complex was centrifuged at 12,000 rpm for another 30 min
and decanted.[16,17] The supernatant was kept and dried in an
oven at 40°C overnight for onward usage.
Sheep red blood cells preparation
Fresh blood was collected from a sheep sacrificed in the local
slaughter house. Sheep red blood cells (SRBCs) were washed
three times in large volumes of pyrogen free 0.9% normal
saline and adjusted to a concentration of 0.5 × 109 cells/ml for
immunization and challenge.[18]
Neutrophil adhesion test
Joshua et al.’s[19] method was employed for neutrophil
adhesion test. Rats of the control group were given 10 ml/
kg normal saline, whereas treatment groups were pre-treated
with different concentrations of nanoparticle extract of P.
merkusii (125; 250 and 500 mg/kg), peroral for 14 days,
respectively. On day 14 of nanoparticle extract of P.
merkusii treatment, blood samples were collected by
puncturing retro-orbital plexus into heparinized vials and
analyzed for total leukocyte cell (TLC) and differential
leukocyte cell (DLC) counts. After initial counts, blood
samples were incubated with nylon fibers for 15 min at
37°C. The incubated blood samples were again analyzed
by TLC and DLC, respectively, to give the neutrophil
index of blood samples. The percent neutrophil adhesion
was calculated by the following formula:
NIu−NIt
Neutrophil adhesion ð%Þ ¼  100;
NIu
where NIu is the neutrophil index of untreated blood samples,
and NIt is the neutrophil index of treated blood samples.
Hemagglutinating antibody titer
Joshua et al.[19] described the procedure for hemagglutinating
antibody (HA) titer. The rats of all groups were immunized by
injecting 0.1 ml of an SRBCs suspension containing 0.5 × 109
cells intraperitoneally on day 0. The rats, which were divided
into control group were given 10 ml/kg normal saline,
whereas treatment groups were pre-treated with different
concentrations of nanoparticle extract of P. merkusii (125;
250 and 500 mg/kg), peroral for 7 days, respectively. Blood
samples were collected from the heart of each rat for
serum preparation on the 7th day. The blood samples were
centrifuged, and serum was obtained. Antibody levels
were determined by the hemagglutination technique. Equal
volumes of individual serum samples of each group were
pooled. Twofold serial dilutions of pooled serum samples
made in 25 ml volume of normal saline in microtitration
plates were added to 25 ml of 1% suspension of SRBCs in
saline. After mixing, the plates were incubated at 37°C for 1 h
11
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Sudjarwo, et al.: The potency of nanoparticle of Pinus merkusii as immunostimulatory
and the value of antibody titer was assigned to the highest
serum dilution showing visible hemagglutination.
Delayed-type hypersensitivity response
The delayed-type hypersensitivity (DTH) response was
determined using the method of Allen.[16] The rats of all
the groups were immunized by injecting 0.1 ml of a SRBCs
suspension containing 0.5 × 109 cells intraperitoneally on
day 0. The rats, which were divided into control group
were given 10 ml/kg normal saline, whereas treatment
groups were pre-treated with different concentrations of
nanoparticle extract of P. merkusii (125; 250 and 500 mg/
kg), peroral for 8 days, respectively. On the 7th day of
immunization, all the rats were challenged with 0.5 × 109
cells in the left hind foot pad. The right footpad was injected
with the same volume of normal saline, which served as the
control for nonspecific swelling. Increase in footpad
thickness was measured 24 h after the challenge.[20]
Phagocytic response (carbon clearance method)
The method was described by Singh et al.[21] The rats, which
were divided into control group were given 10 ml/kg normal
saline, whereas treatment groups were pre-treated with
different concentrations of nanoparticle extract of
P. merkusii (125; 250 and 500 mg/kg), peroral for 7 days,
respectively. On the 7th day, immediate after the last dose
administered to all the rats of each group control as well as
treated received an intravenous injection of carbon suspension
(1:50 dilution of Indian ink, Hi-Media Laboratories Pvt. Ltd.,
Mumbai, India) in a dose of 1 ml/200 g body weight. Blood was
withdrawn from the retro orbital venous plexus before injection
(0 min) and 15 min after injection of the carbon suspension, and
50 ml of blood was lysed with 4 ml of 0.1% sodium carbonate
solution (Na2CO3). The optical density was measured
spectrophotometrically at 650 nm wavelength.
The results were expressed as a phagocytic index:
K ¼ ðln OD12 min Þ−ðln OD0 min Þ=ðt15 min −t0 min Þ;
where OD12 min and OD0 min are the optical densities at time
t15 min and t0 min, respectively.
Cyclophosphamide-induced myelosuppression
The method described by Bin-Hafeez et al.[22] was employed
for cyclophosphamide-induced myelosuppression. Albino
rats were divided into five groups designated as a negative
Table 1: Effect of administration of nanoparticle extract of Pinus merkusii on neutrophil adhesion
Group
UB
Control 331.27 ± 23.35
Pinus merkusii 125 mg/kg 359.16 ± 27.98
Pinus merkusii 250 mg/kg 377.53 ± 31.27
Pinus merkusii 500 mg/kg 458.13 ± 36.62
UB = untreated blood, FTB = fiber treated blood. Values are mean ± SD, n = 6. *P < 0.05 significant.
12 International Journal of Nutrition, Pharmacology, Neurological Diseases ¦ Volume 8 ¦ Issue 1 ¦ January-March 2018
control; positive control and treatment groups, each group
containing six rats. The negative control group received a
saline solution. The positive control group was administered
with only cyclophosphamide at the dose of 30 mg/kg,
i.p. while treatment group rats received cyclophosphamide
and varied concentrations of nanoparticle extract of P.
merkusii (125; 250 and 500 mg/kg, p.o.). The nanoparticle
extract of P. merkusii was given daily for 10 days and
cyclophosphamide was injected with cyclophosphamide on
the 8th, 9th and 10th day, 1 h after the administration of the
respective treatment. Blood samples were collected on the
11th day of the experiment and analyzed for hematological
parameters.
Statistical analyses
Data analyses were performed using the Statistical
Package for the Social Sciences version 12.0 software
for Windows (SPSS Inc., South Wacker Drive, Chicago,
Illinois, USA). All data were expressed as means ±
standard deviation (SD) values. The analysis of variance
test was used to test for differences between the groups.
Duncan’s multiple range test was used to analyze
differences between the mean values and differences
were considered statistically significant at P < 0.05.
RESULTS
Effect of nanoparticle extract of P. merkusii on
neutrophil adhesion
Incubation of neutrophils with nylon fibers produced a decrease
in the neutrophil counts due to adhesion of neutrophils to the
fibers. Effect of nanoparticle extract of P. merkusii on
neutrophil activation by the neutrophil adhesion test is
shown in [Table 1]. The neutrophil adhesion in control
group rats was 5.74 ± 1.72. The nanoparticle extract of P.
merkusii showed a significant increase in neutrophil
adhesion at a dose of 500 mg/kg BW, but not at doses of
125 and 250 mg/kg, when the data were compared with
control group rats, suggesting possible immunostimulant
action of the nanoparticle extract of P. merkusii [Table 1].
Effect of nanoparticle extract of P. merkusii on
humoral immunity parameters
The hemagglutination antibody titer was used to assess
humoral immune response. The humoral antibody titer
value of control group was found to be 31.13 ± 3.85.
Neutrophil index Neutrophil adhesion (%)
FTB
312.14 ± 19.52 5.74 ± 1.72
328.23 ± 26.43 8.63 ± 2.43
336.42 ± 23.78 10.8 ± 3.85
374.19 ± 32.62 18.3 ± 2.27*
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Sudjarwo, et al.: The potency of nanoparticle of Pinus merkusii as immunostimulatory

Administration of nanoparticle extract of P. merkusii doses

Table 2: Effect of administration of nanoparticle extract of
500 mg/kg but not at doses of 125 and 250 mg/kg produced a
Pinus merkusii on hemagglutination titer
significant increase in hemagglutination antibody titer, when
the data were compared with control group rats, as an Group Hemagglutination titer
evidence from hemagglutination after incubation of serum Control 31.13 ± 3.85
with SRBCs [Table 2]. Nanoparticle extract of Pinus 35.21 ± 4.27
merkusii 125 mg/kg BW
Effect of nanoparticle extract of P. merkusii on cell Nanoparticle extract of Pinus 38.23 ± 4.69
merkusii 250 mg/kg BW
mediated immunity parameters
Nanoparticle extract of Pinus 58.75 ± 6.31*
The cell-mediated immune response of nanoparticle merkusii 500 mg/kg BW
extract of P. merkusii was assessed by DTH reaction, that Values are mean ± SD, n = 6. *P < 0.05 significant.
is, foot pad reaction. As shown in Table 3, the nanoparticle
extract of P. merkusii produced a significant, dose-
dependent manner increase in DTH reactivity in rats. The
Table 3: Effect of administration of nanoparticle extract of
nanoparticle extract of P. merkusii at doses 500 mg/kg BW
Pinus merkusii on delayed-type hypersensitivity
but not at dose 125 and 250 g/kg significantly increased the
DTH reactivity as compared to the control. Increase in DTH Group Delayed-type
reaction in rats in response to cell-dependent antigen hypersensitivity 24 h (mm)
revealed the stimulatory effect of nanoparticle extract of Control 0.17 ± 0.05
P. merkusii on T cells [Table 3]. Nanoparticle extract of Pinus 0.21 ± 0.07
merkusii 125 mg/kg BW
Nanoparticle extract of Pinus 0.32 ± 0.09
Effect of nanoparticle extract of P. merkusii on merkusii 250 mg/kg BW
phagocytic response Nanoparticle extract of Pinus 0.78 ± 0.71*
The faster removal of carbon particles has been correlated with merkusii 500 mg/kg BW
the enhanced phagocytic activity. The phagocytic activity of the Values are mean ± SD, n = 6. *P < 0.05 significant.
reticular-endothelial system was measured by the removal of
carbon particles from the blood circulation. The phagocytic
index of the control group was 4.18 ± 0.67 [Table 4]. Oral Table 4: Effect of administration of nanoparticle extract of
administration of nanoparticle extract of P. merkusii a dose- Pinus merkusii on phagocytic index
related increase in the clearance rate of carbon by the cells of the Group Phagocytic index
reticuloendothelial system. The nanoparticle extract of P.
Control 4.18 ± 0.67
merkusii showed significant increase in the phagocytic index Nanoparticle extract of Pinus 4.86 ± 0.91
at doses of 500 mg/kg but not at doses of 125 and 250 mg/kg, merkusii 125 mg/kg BW
when the data were compared with control group rats, Nanoparticle extract of Pinus 5.23 ± 0.89
suggesting phagocytic activity of the nanoparticle extract of merkusii 250 mg/kg BW
P. merkusii. Nanoparticle extract of Pinus 8.75 ± 0.71*
merkusii 500 mg/kg BW
Values are mean ± SD, n = 6. *P < 0.05 significant.
Effect of nanoparticle extract of P. merkusii on
cyclophosphamide-induced myelosuppression
Cyclophosphamide at the dose of 30 mg/kg, i.p. caused DISCUSSION
a significant reduction in the hemoglobin, red blood The immune system is the vital defense against noninfectious
cells (RBCs); white blood cells (WBCs), and platelets count. and infectious diseases. A strong immune system comprises
Combined treatment of cyclophosphamide and the nanoparticle elements that are in balance with one another; if this balance is
extract of P. merkusii a dose-dependent manner result disturbed, our immune system will be incapable to protect the
in a restoration of bone marrow activity as compared body against harmful substances.[2,3] Immunomodulation
with cyclophosphamide treatment alone. Significant using medicinal plants can provide a substitute to conven-
reduction in WBC count was observed in rats treated with tional immunotherapy for a range of diseases, especially
cyclophosphamide alone (positive control) as compared to the when host defense mechanism has to be activated under the
negative control. The nanoparticle extract of P. merkusii at conditions of impaired immune response. There are several
doses 500 mg/kg BB significant increased the levels of diseases where immunostimulant drugs are needed to overcome
hemoglobin, RBCs, WBCs and platelets count as compared the immunosuppression induced by drugs or environmental
to the positive control treated with cyclophosphamide, and it factors. There is a strong necessity of the drugs that can enhance
was observed that nanoparticle extract of P. merkusii at the the immune system to combat the immunosuppressive
doses of 125, 250 and 500 mg/kg, respectively, restored the consequences caused by stress, chronic diseases, and
levels of WBC back to normal [Table 5]. conditions of impaired immune responsiveness. Recently,

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Sudjarwo, et al.: The potency of nanoparticle of Pinus merkusii as immunostimulatory
Table 5: Effect of nanoparticle extract of Pinus merkusii on cyclophosphamide-induced myelosuppression
The haematological parameter
Control Cyclophosphamide 30 mg/kg
RBC (×106/ml) 8.1 ± 0.9 5.1 ± 1.1
Hb (g/dl) 14.7 ± 1.5 8.7 ± 1.3
PCV (%) 58.3 ± 7.3 41.3 ± 5.6
WBC (×103/ml) 7.6 ± 1.1 4.6 ± 1.2
MCV (fl) 63.5 ± 7.3 48.5 ± 4.2
MCHC (g/dl) 37.6 ± 4.8 21.6 ± 5.1
Lymphocyte (%) 67.6 ± 9.3 46.6 ± 7.3
Monocyte (%) 3.7 ± 0.5 1.9 ± 0.7
Neutrophils (%) 1.3 ± 0.3 0.9 ± 0.4
Values are expressed as mean ± SD. The data represent the average from six rats. *Significant difference between the Pinus merkusii groups and the control
group (P < 0.05).
medicinal plants and their products have been commonly
used as immunomodulatory.[5] Though medicinal plants
have been investigated for diverse pharmacologic activities,
the immunostimulatory potential of P. merkusii still remains
unknown.
The results obtained in the present study indicate that
nanoparticle extract of P. merkusii is a potent immuno-
stimulant, stimulating both specific and nonspecific
immune mechanisms. Neutrophils are an important
component of the innate immune system, with the main
role in the clearance of extra­cellular pathogens. Both
localization and neutralization of microorganisms are
functions of neutrophil that are regulated by specific
inflammatory mediators released from the site of infection.
The neutrophil, an end cell unable to divide and with limited
capacity for protein synthesis is, nevertheless, capable
of a wide range of responses, in particular chemotaxis,
phagocytosis, exocytosis and both intracellular and
extracellular killing.[18] In the present study, nanoparticle
extract of P. merkusii evoked a significant increase in
percent neutrophils. This may potentially help in
increasing immunity of body against microbial infections.
The augmentation of the humoral immune response to SRBCs
by nanoparticle extract of P. merkusii, as evidenced by an
increase in the antibody titer in rats indicated the enhanced
responsiveness of T and B lymphocyte subsets, involved in
the antibody synthesis. The high values of HA titer obtained
in the case of nanoparticle extract of P. merkusii have
indicated that immunostimulation was achieved through
humoral immunity. B lymphocytes and plasma cells
function in the humoral immunity component of the
adaptive immune system by secreting antibodies such as
IgG and IgM are the major immunoglobulins which are
involved in the complement activation, opsonization,
neutralization of foreign bodies.[18]
Cell-mediated immunity (CMI) involves effectors
mechanisms performed by T lymphocytes and their
products (lymphokines). CMI responses are critical to
defense against infectious microorganisms, infection of
14 International Journal of Nutrition, Pharmacology, Neurological Diseases ¦ Volume 8 ¦ Issue 1 ¦ January-March 2018
Groups
125 mg/kg Pinus merkusii 250 mg/kg 500 mg/kg
6.5 ± 1.1 6.9 ± 1.3 7.9 ± 0.8*
7.6 ± 2.1 8.4 ± 1.4 13.3 ± 1.8*
45.2 ± 6.5 51.3 ± 8.2 56.9 ± 5.6*
5.1 ± 0.9 6.5 ± 1.3 6.9 ± 0.8*
51.5 ± 6.4 55.5 ± 9.1 61.7 ± 5.9*
24.6 ± 3.9 29.8 ± 5.1 34.1 ± 4.8*
51.5 ± 7.8 56.6 ± 8.9 63.2 ± 8.1*
3.1 ± 0.6 2.9 ± 0.8 3.5 ± 0.6*
1.1 ± 0.5 0.8 ± 0.7 1.2 ± 0.5*
foreign grafts, tumor immunity, and DTH reactions.[2,3]
Therefore, an increase in DTH reaction in rats in response
to T cell-dependent antigen revealed the stimulatory effect of
nanoparticle extract of P. merkusii on T cells. In the present
study, nanoparticle extract of P. merkusii showed an overall
stimulatory effect on the immune functions in rats.
Stimulatory effects were observed on both humoral and
cellular immunity. In DHT test, the nanoparticle extract of
P. merkusii showed an increase response in all doses, but this
increase was significant only in dose 500 mg/kg. This activity
could be due to the presence of pycnogenol that contains
proanthocyanidins which augment the humoral response, by
stimulating the macrophages and B lymphocytes subsets
involved in antibody synthesis. The mechanism behind this
elevated DTH during the CMI responses could be due to
sensitized T-lymphocytes. When challenged by the antigen,
they are converted to lymphoblast and secrete a variety of
molecules including proinflammatory lymphokines, affecting
more scavengers cells to the site of reaction.[3] An increase in
DTH response indicates a stimulatory effect of the plant
which has occurred on the lymphocytes and accessory cell
types required for the expression of this reaction.[4]
Phagocytosis is the process by which certain body cells,
collectively known as phagocytes, ingests and removes
microorganisms, malignant cells, inorganic particles and
tissue debris.[7] Nanoparticle extract of P. merkusii
appeared to enhance the phagocytic function by exhibiting
a dose-related increase in the clearance rate of carbon by the
cells of the reticulo endothelium system.
The administration of nanoparticle extract of P. merkusii
significantly controls the total WBC count, RBCs count,
and hemoglobin and platelets count and also restored the
myelosuppressive effects induced by cyclophosphamide. The
results of the present study indicate that the nanoparticle
extract of P. merkusii can stimulate the bone marrow activity.
The bone marrow a sensitive target particularly to cytotoxic
drugs such as cyclophosphamide. In fact, bone marrow is the
organ most affected during any immunosuppression therapy
with this cyclophosphamide. Loss of stem cells and the
inability of the bone marrow to regenerate new blood cells
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Sudjarwo, et al.: The potency of nanoparticle of Pinus merkusii as immunostimulatory
results in thrombocytopenia and leucopenia.[22] Admini-
stration of the nanoparticle extract of P. merkusii was
found to increase the total WBC count, which was lowered
by cyclophosphamide, a cytotoxic drug, indicating that the
nanoparticle extract of P. merkusii can stimulate the bone
marrow activity.
It could be concluded that nanoparticle extract of P. merkusii
may stimulate both cellular and humoral immune responses.
The nanoparticle extract of P. merkusii not only potentiate
nonspecific immune response but also improve humoral as
well as CMI effectively. The effectiveness of nanoparticle
extract of P. merkusii treated animals in overcoming the side
effects of drug-induced myelosuppression provides sufficient
evidence for balancing and adaptogenic efficacy of the
nanoparticle extract of P. merkusii. Thus, from the results
obtained, it can be concluded that nanoparticle extract of
P. merkusii has therapeutic potential and could be served as
an effective immunomodulatory candidate. Further studies on
the mechanism of action of nanoparticle extract of
P. merkusii, to establish its therapeutic potential for the
prevention of autoimmune diseases are planned in the
laboratory.
Acknowledgements
This study was supported by Ministry of Research,
Technology and Higher Education of the Republic
of Indonesia. Grant No.: 004/Sp2H/LT/DRPM/IV/2017,
April 3, 2017.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
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