Pericarp of Mangosteen Inhibits Nuclear Factor κB’s (NF-κB) Activation and Reduces
Expression of ICAM-1 and IL-6 in Atherosclerosis
Djanggan Sargowo1,2 , Hendarto1, M. Saifur Rohman2
Internal Medicine Department-, 2Cardiology Department- Medical Faculty of Brawijaya University-
Saiful Anwar General Hospital, Malang Indonesia
Background: Atherosclerosis caused more than 50% of all deaths in the developed countries, and major morbidity
in worldwide. Atherosclerosis is widely viewed as an inflammatory disease with hypercholesterolemia being a
dominant underlying risk factor.
Objective: To determine the effect of mangosteen peel in inhibition of NF-κB activation and expression of IL -6
and ICAM-1 in mice with atherosclerosis.
Method: Rattus novergicus wistar strain were divided into 5 treatment groups: 4 groups with high-fat diet and one
group with no high-cholesterol diet. 3 groups of mice with high-fat diet is given a rough skin of the mangosteen
extract 200 mg/kg, 400 mg/kg and 800 mg/kg, and 1 group with a high-fat diet without the extract of mangosteen
peel as a positive control. NF-κB activation and expression of ICAM-1 was tested by immunohistochemistry and IL-
6 protein was measured by ELISA method.
Results: There are significant differences in the activation of NF- κB, and expression of ICAM-1 and IL -6 between
the dyslipidemia mice with nondislipidemia mice (p < 0.05). Dose group 800 mg of mangosteen peel extract has the
highest inhibitory effect on NF-κB’s activation and expression of IL-6. Mangosteen peel extract dose of 400 mg and
800 mg inhibit the expression of ICAM -1 and NF-κB significantly.
Conclusion: The mangosteen peel extract significantly inhibited NF-κB’s activation and reduce expression of
ICAM -1 and IL -6 in mice with atherosclerosis.
Keywords : Atherosclerosis, Xanthone, NF-κB, ICAM-1, Interleukin-6
Background inflammatory disease with hypercholesterolemia
Atherosclerosis, the major cause of death being a dominant underlying risk factor. It is believed
from cardiovascular disease in industrialized to be initiated by retention of LDL particles in the
countries (1-2), is characterized by the progressive lesion-prone areas, which is followed by monocyte
accumulation of lipid and fibrous depositions in the recruitment and their differentiation into cholesterol-
vessel wall of large arteries.(2) laden macrophage foam cells. Excessive cholesterol
Despite considerable therapeutic advances accumulation in macrophages exaggerates innate
over the past 50 years, cardiovascular disease is the immune response that is manifested by upregulated
leading cause of death worldwide. This is mainly a production and secretion of inflammatory cytokines
result of the increasing prevalence of atherosclerosis, and chemokines, thus dramatically amplifying initial
owing to the ageing population, the improved signal originated from the injured artery. (4)
survival of patients with atherosclerotic This requires research on the molecular
cardiovascular disease and, above all, the widespread pathophysiology and the incidence of atherosclerosis
under-recognition and undertreatment of individuals process, as well as the impact progressivity
with risk factors for atherosclerosis. (3) complications, so it can be known with certainty the
Atherosclerosis is a disease of large arteries mechanism of action in which the drug works. One
that accounts for more than 50% of all deaths in the alternative that is very good as a cure at this point is
developed countries and, according to the World to start many examined the alleged benefits of
Health Organization prediction, it will gain a status of mangosteen peel as an anti-sclerotic effects because it
the major morbidity cause worldwide by the year proved to contain xanthone, which only contained a
2010. Atherosclerosis is widely viewed as an substance on the peel as an anti- oxidant mangosteen.
Recently, the pharmacological properties of Results
xanthones in the cardiovascular system have attracted Figure 1. In the negative control group (A)
great interest. Xanthones and xanthone derivatives occurs the expression of NF-κB activation (arrow),
have been shown to have beneficial effects on some the positive control group who received high-
cardiovascular diseases, including ischemic heart cholesterol diet (B) increased expression of NF-κB
disease, atherosclerosis, hypertension and activation (arrows). On treatment with mangosteen
thrombosis. The protective effects of xanthones in the peel extract dose of 200 mg/kg (C) is still there
cardiovascular system may be due to their appeared to be increased expression of NF-κB
antioxidant, antiinflammatory, platelet aggregation activation. But at a dose of 400 mg / kg (D) and the
inhibitory, antithrombotic and or vasorelaxant dose of 800 mg / kg (E) began to appear decreased
activities. In particular, the antagonism of expression of NF-κB activation.
endogenous nitric oxide synthase inhibitors by Figure 2. In the negative control group (A)
xanthones may represent the basis for improved occurs the expression of ICAM-1 (arrow), the
endothelial function and for reduction of events positive control group who received high-cholesterol
associated with atherosclerosis. (5) diet (B) increased expression of ICAM-1 (arrows).
Based on the work of xanthone found in On treatment with mangosteen peel extract dose of
mangosteen’s skin (Garcinia mangostana) inhibits 200 mg/kg (C) is still there appeared to be increased
inflammatory processes through the transcription expression of ICAM -1. But at a dose of 400 mg/kg
factor NF-κB, we want to prove whether the peel of (D) and the dose of 800 mg / kg (E) began to appear
the mangosteen xanthone in dyslipidemia mice that decreased expression of ICAM -1.
can inhibit NF-κB activation and therefore reduces Table 1. In the negative control group
the expression of IL -6 and ICAM -1. occurred expression of NF-κB activation with the
lowest value (29.67), whereas the positive control
Methods group who received high-cholesterol diet increased
The animal experimental trial study design, the mean expression of NF-κB activation, with the
was conducted at Brawijaya University highest yields (71.83). On treatment with mangosteen
Pharmacology Laboratory, between September and peel extract dose of 200 mg/kg was still an increase
November 2010. A total of 30 subjects, Rattus in the average expression of NF-κB activation
novergicus wistar strain divided into 5 treatment (71.00), and with increasing doses starting to look a
groups; 4 groups with high fat diet and 1 group of decrease in the average expression of NF-κB
non high cholesterol. 3 groups of mice treated with activation, at doses of 400 mg/kg (56.67) and at doses
200 mg/kg, 400 mg/kg and 800 mg/kg of of 800 mg/kg (39.00).
mangosteen pericarp crude extract with 1 group of
high fat diet without treatment as a control positive. Table 1. Mean NF-κB’s activation in aortic endothelial cells
After 12 weeks treatment, NF-κB’s activation and
ICAM-1’s expression was checked by
immunohistochemistry and IL-6 protein was
measured by ELISA methods.
Data analysis, which were performed on Table 2. In the negative control group
One way ANOVA, and continued with Post Hoc occurred expression of ICAM -1 with the lowest
Analyses, to examine the effect of mangosteen’s value (20.50), whereas the positive control group
pericarp with variety doses about NF-κB activation who received high-cholesterol diet increased the
and ICAM-1 expression and IL-6 protein level on mean expression of ICAM -1, with the highest yields
high cholesterol diet treatment group, significance (51.33). On treatment with mangosteen peel extract
level 0.05. Analyses were performed with SPSS dose of 200 mg/kg was still an increase in the mean
version 17.0 for Windows (SPSS, Inc, Chicago, expression of ICAM-1 (45.67), and with increasing
Figure 1. A, B, C, D, E Activation of NF- Figure 2. A, B, C, D, E Activation of
κB Expression in aortic endothelial cells ICAM-1 Expression in aortic endothelial
doses starting to look a decrease in the mean mg/kg obtained average levels of IL-6 is 103.2 pg/ml,
expression of ICAM-1, at doses of 400 mg/kg (37.50) at a dose of 400 mg/kg (249.97 pg/ml) and at doses
and at doses of 800 mg/kg (26.67). of 800 mg/kg (215.67 pg/ml).
Table 3. The mean levels of IL-6 (pg/ml) in serum
Table 2. Mean ICAM-1’s expression in aortic endothelial cells
Table 3. In the negative control group Effect of high dietary cholesterol on the activation
gained an average blood levels of IL-6 is 94.31 pg/ml of NF- κB in the positive control than the negative
and IL-6 levels in the positive control group who control
received high-cholesterol diet that is 93.98 pg/ml. On After the test conducted by One way
treatment with mangosteen peel extract dose of 200 ANOVA and Post Hoc analysis, as found in figure 3
obtained the result that there are significant treatment groups on the expression of NF-κB
differences in the expression of NF-κB activation activation, expression of ICAM-1 (p-value = 0.000)
between the mice fed a cholesterol diet [ K (+) ] with and levels of IL -6 (p-value = 0.016).
mice not given a diet high in cholesterol [K(+) 71.83; Based on the results of Post Hoc test seen
K(-) 29.67; p = 0.000]. that the addition of mangosteen peel extracts on the
mice fed high-cholesterol diet lowers the expression
of NF-κB activation and ICAM-1. Dose of 400
mg/kg of mangosteen peel extract has the effect of
decreased expression of NF-κB (p-value = 0.000)
and ICAM-1 (p-value = 0.000) when compared with
positive control group. Dose of 800 mg/kg of
mangosteen peel extract has the effect of decreased
expression of NF-κB (p-value = 0.000) and ICAM -1
(p-value = 0.000) and decreased levels of IL -6 (p-
p = 0.000 value = 0.004) when compared with positive control
Thus concluded that the addition of
Figure 3. Graph of the effect of high dietary cholesterol on the mangosteen peel extract of 800 mg/kg in mice fed a
activation of NF-κB in aortic endothelial cells. Data expressed as high cholesterol diet is the best dose to reduce the
average number, n = 6, meaning if the value of p <0.05 expression of NF-κB’s activation, expression of
ICAM-1, and IL-6 levels, although the inhibitory
In figure 4 obtained the result that there are effect of ICAM -1 expression and activation NF- κB
significant differences in ICAM-1 expression has been significant with a dose of 400 mg/kg with
between mice fed a cholesterol diet [ K (+) ] with significant results.
mice not given a high cholesterol diet [ K (-) ], [K(+)
51.33; K(-) 20.50; p = 0.000]. Discussion
Atherosclerosis is a chronic disease of the
arterial wall where both innate and adaptive
immunoinflammatory mechanisms are involved.
Inflammation is central at all stages of
atherosclerosis. It is implicated in the formation of
early fatty streaks, when the endothelium is activated
and expresses chemokines and adhesion molecules
leading to monocyte/lymphocyte recruitment and
infiltration into the subendothelium. It also acts at the
p = 0.000 onset of adverse clinical vascular events, when
activated cells within the plaque secrete matrix
proteases that degrade extracellular matrix proteins
Figure 4. Graph of the effect of high dietary cholesterol on the and weaken the fibrous cap, leading to rupture and
activation of ICAM-1 in aortic endothelial cells. Data expressed as
thrombus formation. Cells involved in the
average number, n = 6, meaning if the value of p <0.05
atherosclerotic process secrete and are activated by
soluble factors, known as cytokines. (6)
Effect of various doses of mangosteen peel extracts
LDL oxidation has a key role in the
on NF - κB activation and ICAM -1 expression in
activation and facilitation of atherogenesis. The
aortic endothelial cells of mice that were given
signals in the form of oxidized lipoproteins and
reactive oxygen species (ROS) lead to the activation
of transcription factors such as nuclear factor-kB
Based on the ANOVA test shows that there
(NF-κB) that are involved in the regulation of
is a significant difference in effect between the five
expression of ICAM-1. (7)
Figure 5. Graph Effect of mangosteen peel extract on NF-κB’s activation in endothelial cells
aorta rats fed high-cholesterol diet. Data expressed in mean number of endothelial cells
expressing NF-κB activation, n = 6, significant when the p-value <0.05
Figure 6. Graph Effect of mangosteen peel extract on the expression of ICAM -1 on endothelial cells
aorta rats fed high-cholesterol diet. Data expressed in mean number of endothelial cells
expressing ICAM -1, n = 6, significant when the p-value <0.05.
Invitro studies had explained that activation of activation can stimulate ROS production and activate
oxLDL-mediated endothelial LOX-1-linked signaling nuclear factor κB (NF-κB). Activation of NF-κB and
pathway causing pro-inflammatory respons. LOX-1 translocation into cell nucleus, result in increase of
pro-inflammatory and adhesion molecules coding expression of ICAM-1 in mice fed a high cholesterol
genes, i.e. tumor necrosis factor-α, ICAM-1, and diet with mangosteen peel extract in various doses.
VCAM-1. (8) The 400 mg and 800 mg dose of mangosteen peel
can inhibit NF- κB and ICAM-1 significantly.
Several studies have shown that antioxidant
nutrients and (or) natural medicine positively
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