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Pericarp-of-Mangosteen-Inhibits-Nuclear-Factor-κB’s-NF-κB

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					   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
   1
       Internal Medicine Department-, 2Cardiology Department- Medical Faculty of Brawijaya University-
                              Saiful Anwar General Hospital, Malang Indonesia
Abstract
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.


                                                                                                                   1
          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.

Analysis
           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
Illinois).

                                                                                                                        2
              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
                                                                                   cells
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

                                                                                                                    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
                                                                      group.
                                                                                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
high-cholesterol diet
                                                                      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)

                                                                                                                             4
                    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


                                                                                                                           5
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
modulate the susceptibility of LDL to oxidation and          References
enhance the antiatherogenic properties of HDL, thus              1. Kobayashi K, Inoguchi T. Molecular
playing an important role in the prevention of                      Mechanisms of the Antiatherogenic Action
cardiovascular diseases. (9)                                        of     Adiponectin.     Vascular    Disease
         Many studies have shown that free radicals                 Prevention. Vascular Disease Prevention.
cause oxidative damage to fats, and nucleic acids.                  2007;4(1):7-9.
Antioxidants seem very important in the prevention               2. Zandbergen F, Plutzky J. PPARα in
of degenerative diseases ( including heart disease and              atherosclerosis and inflammation. Biochim
blood vessels ), because it can inhibit the formation               Biophys Acta. 2007 August;1771(8):972-82.
of the substrate chain reaction that can be oxidized             3. Brand K, Page S, Walli AK, Neumeier D,
(10)
                                                                    Baeuerle PA. Physiological Society
          Mangostin contained in mangosteen peel,                   Symposium: Impaired endothelial and
that is an effective inhibitor of LDL peroxidation in               smooth muscle cell function in oxidative
vitro as indicated by lagtime to oxidation and                      stress Role of nuclear factor-κB in
malondialdehyde production. Derivatives of the                      atherogenesis     Experimental Physiology.
parent compound have been produced and may offer                    1997;82:297-304.
further benefits and therapeutic potential. (11)                 4. Frolov A, Hui DY. The Modern Art of
          There is substantial evidence to suggest that             Atherosclerosis: A Picture of Colorful
xanthones and xanthone derivatives may be                           Plants, Cholesterol, and Inflammation.
potentially useful as pharmacological agents in the                 Arterioscler     Thromb      Vasc     Biol.
treatment or prevention of cardiovascular diseases,                 2007;27:450-2.
including ischemic heart disease, atherosclerosis and            5. Jiang D-J, Dai Z, Li Y-J. Pharmacological
hypertension. The protective effects of xanthones in                Effects of Xanthones as Cardiovascular
the cardiovascular system may be due to their                       Protective Agents. Cardiovascular Drug
antioxidant, anti-inflammatory, platelet aggregation                Reviews. 2004;22(2):91-102.
inhibitory, antithrombotic andor vasorelaxant                    6. Ozturk Z, Sonmez H, Gorgun FM, Ekmekci
activities. In particular, the antagonism of                        H, Bilgen D, Ozen N. The Relationship
endogenous NOS inhibitors by xanthones may                          Between Lipid Peroxidation and LDL
represent the basis for improved endothelial function               Desialylation        in        Experimental
and for reduction of events associated with                         Atherosclerosis. Toxicology Mechanisms
atherosclerosis. However, the precise effects of                    and Methods. 2007;17:265-73.
xanthones need to be further elucidated in animal                7. Fotie J, Bohle DS. Pharmacological and
experiments in vivo and in humans. Moreover,                        Biological Activities of Xanthones. Anti-
pharmacokinetics,       toxicity     and        structural          Infective Agents in Medicinal Chemistry.
optimization of xanthones should also be explored. (5)              2006;5:15-31.
                                                                 8. Vohra RS, Murphy JE, Walker JH,
Conclusion                                                          Ponnambalam S, Homer-Vanniasinkam S.
                                                                    The LOX-1 Scavenger Receptor And
         There is an increased activation of NF-kB                  Atherosclerotic Plaque Rupture. Trends
and increased expression of ICAM-1 significantly (p                 Cardiovasc Med. 2006;1:60-4.
< 0.05) in mice fed a high cholesterol diet ( K + )              9. Nakatani K, Yamakuni T. γ-Mangostin
compared to negative control ( K - ). There are                     Inhibits Inhibitor-κB Kinase Activity and
inhibition to activation of NF - κB and decreased                   Decreases       Lipopolysaccharide-Induced

                                                                                                              6
    Cyclooxygenase-2 Gene Expression in C6
    Rat Glioma Cells. . Mol Pharmacol.
    2004;66:667-74.
10. Tedgui A, Mallat Z. Cytokines in
    Atherosclerosis: Pathogenic and Regulatory
    Pathways. Physiol Rev. 2006;86:515-81.
11. Williams P, Ongsakul M, Proudfoot J, Croft
    K, Beilin L. Mangostin inhibits the oxidative
    modification of human low density
    lipoprotein. Free Radic Res. 1995
    Aug;23(2):175-84.




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