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The Effect of Cinnamomum burmannii Extract as an Immunomodulator on the

Increase of GR-1 Expressing IFNγ and Macrophage

Article · October 2015

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The Effect of Cinnamomum burmannii Extract as an Immunomodulator on
the Increase of GR-1 Expressing IFNγ and Macrophage
Pratiwi TS 1, Putri A2, Murwani S3
1
Professor of Animal Husbandry Faculty, University of Brawijaya, Malang-65145,
Indonesia, 2 Student of Medical Veterinary School, University of Brawijaya, Malang-65145,
Indonesia, 3.Lecturer of Medical Veterinary School , University of Brawijaya Malang,
Indonesia 65145

ABSTRACT
Cinnamomum burmannii was known as one of herbal medicine that has been used
traditionally as an ingredient of traditional medicine extract, contains cinnamaldehyde,
which is a naturally trigger of the body's immune response. This study aimed to evaluate the
immunostimulant effect of C. Burmannii and the increase of Granulocyte Receptor - 1 (GR-
1) that can be recognized from neutrophil expressing IFNγ and the increase of macrophage
phagocytosis activities.Thirty wistar mouse were appropriatelly infected with Salmonella
enteridis and then orally treated with C.burmanni alcohol extract at different dosages. The
mouse were observed for the increase of GR-1 expressing IFNγ using flow-cytometry and
the increase of macrophage phagocytosis activities using Giemza staining. The result showed
that increasing dosage of C. burmannii extract treatment increased the GR-1 level and the
IFNγ by 97.7% and macrophage phagocytosis activity by 98.1% (P<0.05). Thus, the
observations reflected clearly that C.burmannii ethanol extract can be utilized as
immunomodulator that increased immune response.

Keywords: Cinnamomum burmannii, GR-1, IFNγ, macrophage, phagocytosis

Corresponding author : pratiwi_trisunuwati@yahoo.com

INTRODUCTION models of Mus musculus. Several

Burmannii cinnamon of Cinnamomum
species is one of the medicinal plants that
are often found in regions of Indonesia
(Gunawan and Mulyani, 2004), contains
some compounds that are beneficial to
health including drug efficacious for gout,
high blood pressure. Arrar (2009) reported
that C burmannii was also proven to be
antibacterial in Bacillus cereus, Listeria
monocytogenes, Staphylococccus aureus,
Helicobacter pylori, Salmonella
typimurium, and Escherichia coli. Active
compounds contained in cinnamon is
essential oil, safrole, sinamadehid,
eugenol, tannins, resins, calcium oxalate,
tanning agents, flavonoids, saponins and
other nutritional content such as sugar,
protein, crude fat and pectin which
allegedly helped in immune response
(Gunawan and Mulyani, 2004; Guanther,
2006; Wang, 2009). Ramchandra (2006)
reported that cinnamon of Cinnamomum
zeylanicum species at a dose of 100 mg /kg
had immunostimulatory effects in animal
researchs on herbs to enhance immune
responses have been carried out, for
example research on the effect of extracts
of Hedyotis corymbosa, Cassia Alata L
leaf extract, Blumea balsamifera on
phagocytic activity of macrophages Mus
musculus strain BALB/c that were infected
by Salmonella typhimurium, Listeria
monocytogenes on the efect of
imunomudulatory (Kusmardi, 2007);
(Munawaroh, 2008) on macrophage
phagocytic potential. Dewanti et al (2010)
examined the effect of aqueous extract
Grass Gelata against interferon gamma
expression and activity of NK cells,
cytotoxic cells and macrophages in vivo to
determine the effect its cytotoxicity using
myeloma cell culture Mus musculus.
However, research on the
immunomodulatory effects of ethanol
extract Cinamomum of Cinamomum
burmannii to increase phagocytic activity
of interferon gamma and macrophages on
Mus musculus strain BALB/c has not yet
been done before and needs further
research. T cells plasma in the immune
system produce IFNγ is a specific activator
of macrophage function and plays a
critical role in the host immune defense to
bacterial infection that activate
macrophages as a system of mononuclear
phagocytes and polymorphonuclear
neutrophils as phagocyte system (Abbas,
2007)
MATERIAL AND METHODS
Thirty male wistar BALB/c (SPF) were
appropriatelly infected with Salmonella
enteridis at a dose of 0.2 ml x 108 ml/CFU
and then orally treated along 14 days with
C.burmanni ethanol (high polar) extract at
different dosages i.e 0, 50, 100,150 and
200 mg/kg body weight (P0, P1, P2,, P3,
P4). Samples of peritonial fluid and spleen
tissue were collected, one drop of
peritonial fluid sample as a thin smear
Giemza staining test to evaluate the
number of active macrophages in activity
phagocytosis. While the mouse were
observed for the increase of GR-1
expressing IFNγ using flow-cytometry
(Rifa'i, 2004). Analysis of data on the
number GR-1 that express IFNγ and
phagocytic activity of macrophages using
one-way ANOVA.
RESULTS AND DISCUSSION
Based on the number of CD4 that
expressed IFNγ (bound to the extracellular
antibody GR-1), it was shown that mice
treated using C. burmanii ethanol extract
showed significantly higher (p<0.05)
average number of CD4 that expressed
IFNγ compared to those in either the
negative control treatment or positive
control treatment (Tab.1 and Fig.1). While,
positive control treatment (P0+) showed
also significantly higher (p<0.05) average
number of CD4 that expressed IFNγ than
negative control treatment (P0-). The data
were in line with research conducted by
Eckmann (2001) and Lestarini (2008).
They indicated that number of neutrophil
cells (CD4) and IFNγ was higher in
animals infected with Salmonella
enteritidis than in non-infected. In
addition, interferon gamma (IFNγ)
regulate gene expression in a number of
neutrophils including complement receptor
regulator, B lymphocyte stimulator,
dendrites chemotactic factors, chemokines
receptors, neutrophil chemotactic factors
and pro-inflammatory cytokines (Roilides
et al., 2002).Sinamaldehid inhibits the
release of ROS and activates MAPKs as an
expression of pro-inflammatory cytokines.
Finally express highly increased against
GR-1(IFNγ) and the number of active
macrophage cells and regulate gene
expression against complement regulator
B cells and proinflamatory cytokines. .

Table 1. Average number of CD4 in Salmonella enteritidis non-infected and infected and cinnamon (C.
burmanii) ethanol extract non-treated and treated animals
Number of CD4 (cells)
Treatment
Replic. 1 Replic. 2 Replic. 3 Replic. 4 Average + Sd
P0- 314.025 306.000 303.400 300.900 306.081,25 ± 5.690,46a
P0+ 1.587.425 1.551.000 1.428.875 1.514.750 1.520.512,50 ± 67.915,16b
P1+ 2.034.875 1.979.125 1.482.800 2.057.000 1.888.450,00 ± 272.411,00b
P2+ 2.554.375 2.593.500 2.672.325 2.013.725 2.458.481,25 ± 30.0534,49c
P3+ 3.243.275 2.662.375 3.329.100 3.185.925 3.105.168,75 ± 301.001,89d
P4+ 3.933.000 3.932.500 4.024.125 3.864.500 3.938.531,25 ± 65.507,98e
Notes: Values in the same column with different superscript showed significantly difference (P <0.05)
Figure 1. IFNγ as measured (flowcytometry) of mice treated C. Burmanii ethanol extract
showed significantly higher (p<0.05) than those in either the negative control
treatment or positive control treatment

Macrophages Phagocytosis Activity Fig.2 Phagocytosis activity of active

Phagocytic activity of macrophages were macrophage cells against S.
calculated from peritonial fluid smears test enteritidis
(Fig.2), average number of active
macrophage cells that performed
phagocytosis in Salmonella enteritidis
non-infected and infected and cinnamon
(C. burmanii) extract non-treated and
treated animals was presented in Tab. 2. It
was found that average active macrophage
cells that performed phagocytosis
increased linearly (P <0.05) with the
increase of level of cinnamon (C.
burmanii) ethanol extract treatment.

Table 2. Average number of active macrophage cells that performed phagocytosis in Salmonella enteritidis
non-infected,infected and cinnamon (C. burmanii) treatment
Number of active macrophage cells
Treatment
Replic. 1 Replic. 2 Replic. 3 Replic. 4 Average + Sd
P0- 46 47 42 39 43.50±3.69a
P0+ 55 56 58 61 57.50±2.64b
P1+ 69 63 64 65 65.25±2.62c
P2+ 74 73 79 81 76.75±3.86d
P3+ 86 84 85 83 84.50±1.29e
P4+ 87 91 93 96 91.75±3.77f
Notes: Values in the same column with different superscript showed significantly difference (P <0.05)
CONCLUSION
C. burmannii extract treatment on S. characterized by the increase of GR-1
enteritidis infected M musculus BALB/c expressing IFNγ and the number of active
have potential as immunostimulatory as phagocyte macrophage cells.

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