Supplementation of Cantharanthus Roseus Leaf Extract as Anti-Inflammatory Substance to Hypercholesterolemic Wistar Derived-Rats.
Pure and Applied Chemical Sciences, Vol. 3, 2015, no. 1, 1 - 9
HIKARI Ltd, www.m-hikari.com
http://dx.doi.org/10.12988/pacs.2015.41011
Supplementation of Cantharanthus Roseus Leaf
Extract as Anti-Inflammatory
Substance to Hypercholesterolemic Wistar
Derived-Rats
Sri Wahjuni1, Sri Rahayu Santi1 and A. A Ngurah Gunawan2
1
Department of Chemistry
Faculty of Mathematics and Natural Science
Udayana University, Bali, Indonesia
1
Department of Chemistry
Faculty of Mathematics and Natural Science
Udayana University, Bali, Indonesia
2
Department of Physics
Faculty of Mathematics and Natural Science
Udayana University, Bali, Indonesia
Copyright © 2014 Sri Wahjuni, Sri Rahayu Santi and A. A Ngurah Gunawan. This is an open
access article distributed under the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited.
Abstract
The present study on supplementation of Cantharanthus roseus leaf extract
(CRLE) to rats suffer from hypercholesterolemia was aimed to determine whether
it has property as an anti-inflammatory substance assessed from concentrations of
tumor necrosis factor-α (TNF- α) and Interleukin-6 (IL-6) in the blood serum. A
randomized pre-test and post-test design was employed, using 4 weeks old Wistar
derived-rats at body weight ranging from 70 – 72 gram. The rats were initially fed
with normal rat pellets for 1 month followed by high fat ration for 6 weeks in
order to induce hypercholesterolemia condition. Then they were divided into 5
groups according to various treatments applied as follows. Group 1: control –
2
Sri Wahjuni et al.
supplemented with 0% CRLE; Group 2: 10% CRLE; Group 3: 15% CRLE; Group
4: 20% CRLE; and Group 5: 25% CRLE. CRLE supplementation was last for 1
month. TNF- αconcentration was determined using Sandwich Enzyme (ELISA)
kit spectrophotometer (at π =500 nm) andconcentration of IL-6 was determined
using Quintikine HS immunoassay kit. In general, results of the current study
showed that CRLE may reduce inflammation based on assessment of
concentrations of TNF- α and IL-6 in the serum. Following supplementation of
20% CRLE, TNF- α concentration decreased from 28.62 pg/ml to 15.56 pg/ml
while IL-6 decreased from 134.64 g/ml to 113.87 pg/ml. In Indonesia,
Cantharanthus roseus grown as ornament plant and contain tannin, tripernoids,
and alkaloids. The component that acts an anti-inflammation is flavanoids.
Keywords: Leaf of Cantharanthus roseus, tumor necrosis factor (TNF),
interleukin-6 (IL-6), anti-inflammation, ELISA
Introduction
In Indonesia, research work on traditional medicine is still preferable for
combating animal diseases and human diseases. One of herbal plants that have
been proved beneficial for curing various diseases is Cantharanthus roseus (C.
roseus). It grown as ornament plants and its local name is “tapak dara ”. As
reported from many countries such as India and China, extract of C. roseus leaf
can be used to cure many diseases (Singh and Singh, 2001). It has been
discovered that extract leaf of C. roseus contains tannin, trypernoid, alkaloid, and
flavonoids. Flavonoids occur in the form of glycoside and it has been shown in
vivo that flavonoids may inhibit activity of NF-kB (nuclear factor kappa-B),
leading to inhibition of inflammation reaction (Han et al., 2007).
Inflammation is a complex condition that starts from tissue damaging.
Damage of the tissues are due to endogen factors (such as tissue necrosis) and to
external factors such as contact with foreign particles or infections. Inflammation
is generally a kind of response of vascular living tissues, that followed by
important endothelium process. Process occurring in the endothelium of blood
vessel lead in turn to development of atherosclerosis (Hosaya et al.,
2005).Atherosclerosis may become the consequence of hypercholesterolemia, also
called dyslipidemia – the increase in lipid content in the serum. Low density
lipoprotein (LDL) particularly the oxidized form may significantly cause damage
of the endothelium. On the other hand, high density lipoprotein (HDL) is the
protective component (Robbin and Cataren, 2002). Furthermore, chronic
hypercholesterolemia may result in cardiovascular diseases.
Prior to the occurrence of atherosclerosis and the subsequent diseases,
inflammation occurs. Thus, attempts must be made to reduce or minimized
inflammation reaction in order to avoid or minimize the effects of such various
health problems. For such aims, the current research work is designed to study the
Supplementation of Cantharanthus roseus leaf extract
3
effect of supplementation of extract leaf of herbal plant C. roseus as antiinflammatory substance, based on determination of pro-inflammatory cytokines
(TNF- α and IL-6) in the serum of experimental rats that underwent
hypercholesterolemia.
Materials and Methods
Materials
Animals used in the present study are: 50 male Wistar derived-rats at 4
weeks of age and weighing 70 – 72 grams. Experimental materials include C.
roseus leaf extract (CRLE), Sandwich enzyme immunoassay (ELISA) kit for
determination of TNF- α, Quintikine HS immunoassay kit for IL-6 determination,
EDTA, and diethyl ether. Serum collected from the experimental animals were
kept at -200C until assays were conducted. Prior to assays, serum samples were
centrifuged at 2000 rpmfor 10 minutes.
Methods
Experimental design employed was a randomized pre-test and post-test.
The experimental animals were divided into 5 groups according to the treatment
applied, namely Group 1 (control): supplemented with 0% CRLE; Group 2: 10%
CRLE; Group 3: 15% CRLE; Group 4: 20% CRLE; and Group 5: 25%
CRLE.Prior to supplementation of CRLE, all experimental animals were initially
raised under normal rat pellets for 1 month. Then, they are all subjected to high fat
ration for 6 weekin order to make them suffering from hypercholesterolemia.Such
feeding treatment has been shown to result in hypercholesterolemia and chronic
inflamationin rats (Wahjuni,2011). At the end of the high fat feeding phase,
concentrations of TNF-α and IL-6 were measured and the values were considered
as the pre-test data. Supplementation of the various dosages of CRLE to the
experimental animals lasted for 6 weeks. At the end of this phase, concentrations
of TNF-α and IL-6 were again measured and the data was considered as the posttest values
The data was analyzed descriptively. Data selection including editing,
coding and tabulation was performed using Windows file navigator statistical
program (Triton, 2006; Pramesti, 2007). For normality analysis of TNF-α and IL6 data, Shapiro Wilk test was employed at 5% level of significant (p = 0.05). H0
hypothesis: frequency of observation/ frequency of expectation. Based on this
test, the data was considered to be normally distributed when P > 0.05. Moreover,
variant homogeneity was analyzed using the Levent’s test.Ho hypothesis:
variation of control group = variation of Group 1 = variation of Group 2 =
variation of Group 3 = variation of Group 4. Ho was accepted when P > a, and
vice versa .
4
Sri Wahjuni et al.
Result and discussion
Decrease in TNF-α concentration
The average level of TNF-α measured from blood of 50 male rats suffering
from hypercholesterolemia at pre-test and post-test was presented at Table 1. The
pre-test TNF-α values for control group, Group 1, Group 2, Group 3 and Group 4
were (28.98 ± 6.00); (29.12 ± 5.79);(29.02 ± 5.34); (28.62 ± 4.72); dan (29,02 ±
5.06) pg/mLrespectively. Meanwhile, the post-test values for control group,
Group 1, Group 2, Group 3 and Group 4 were (28.11 ± 5.94); (27.32
±5.01);(24.42 ± 5.74); (15,56 ±7,20); ( 26,02 ± 8,34 )pg/mL respectively. When
the pre-test and post-test values were compared, there was no significant reduction
in concentration of TNF-α for the control group. On the other hand,
supplementation of CRLE at 10%, 15%, 20% and 25% has resulted in decreasing
level of TNF-α. Supplementation of CRLE at 10% and 15% led to reduction of
TNF-α level up to 0.79 and 3.69 pg/ml, respectively. However, statistical analysis
showed that such differences were not significant (P> 0.05). But a significant
difference (P < 0.05) in TNF-α concentrations between the pre-test and post-test
values was noted for rats subjected to supplementation of 20% CRLE (Group 3).
For this group, TNF-α concentration has decreased at amount of 12.55 pg/ml.
Furthermore, for rats under 25% CRLE supplementation, no significant different
(P > 0.05) in TNF-α concentration was observed.
Tablel 1
Decrease level of TNF-α serum Wistar Rat Hiperkolesterolemia
Treatment
Observation TNF-α (pg /mL)
Pre-test
Pos-test
EDGR 0 % (control)
EDGR 10%
EDGR 15%
EDGR 20%
EDGR 25%
28.98 ± 6.00
29,12± 5,79
29,02 ± 5,34
28,62 ± 4,72
29,02 ± 5.06
28.11 ± 5.94
27,32 ± 5,01
24,42 ± 5,74
15,56 ± 7,20
26,02 ± 8,34
The present study showed that supplementation of 20% CRLE has resulted
in the highest reduction in the serum TNF-α concentration. But supplementation
of CRLE at 25% did not lead to further decrease in TNF-α concentration. It seems
that at 25% supplementation, the response has reached saturated state, so that
cannot cause further decreasing effect on TNF-α concentration. The present result
on supplementation of 25% CRLE seems in accordance with the findings of Chen
and Goeddel (2002) who reported that TNF-αproduced by NF-KB do not undergo
transcription reaction when it reaches saturated state. Therefore, transcription
factors are responsible for induction of gene expression for inflammation process
to occur.
Supplementation of Cantharanthus roseus leaf extract
5
Inflammation is generally a response to external and internal factors by
vascular living tissues. The inflammation response is usually followed by an
important process – endothelium process. Endothelium of blood vessel is the site
where the deposition of fat occurs and known as atherosclerosis. Endothelium is
the primary target for mechanical and chemical injuries caused by
hypercholesterolemia. Chronic hypercholesterolemia has been known to result in
the shift of acute pro-inflammation and prothrombosis responses into the chronic
responses. These responses, in turn, are followed by infiltration of leucocytes,
particularly of monocyte, to sub-endothelium, and subsequentlychanging into
macrophage cells. The macrophage cells then phagocyte the remnant of LDL
which is oxidized to form atheroma (Baraas, 2006).
Simopolous (2002) reported that flavonoids as anti-inflammation play role
as immunomodulator. Mechanismeis increas antioxtidan endogen and capcing
free radikal.Inhibition stres oksidatif by some flavonoid incontain of
cantharanthus roseus Leaf Extract (Figure 1). And than flavonoid cantharanthus
roseus isolasitian metabolit sekunder compound, extraction, distilation,
purification, dan identificationby GC-MS(Gas Chromatograph Mass
Spectrometer ) Shimadzu QP2010, identifay flavonoid what typeCantharanthus
roseus Leaf Extract.
Figure 1: Habition Flavonoid 0f NF-kB (Nuclear Factor Kappa Beta)was decreas
habition
interleukin product pro inflamation, COX2 can habition reaction
inflamation.
Its role as anti-inflammation is due to the weakening effect of arachidonic acid.
Arachidonic acid may activate cyclooxygenase and 5-lipooxygenase enzymes.
These two enzymes act as endothelium-dependent vasodilator that cause
relaxation of normal coronary arteries and cause paradox vasoconstriction of
arteries in hypercholesterolemia condition.
6
Sri Wahjuni et al.
Decrease in IL-6 concentration
Concentrations of interleukin-6 (IL-6) in blood serum of rats suffering
from hypercholesterolemia and then treated with CRLE at pre-test and post-test
are presented in Table 2. The pre-test IL-6 concentrations for group supplemented
with 0% CRLE (control), with 10%, 15%, 20%, and with 25% CRLE were
(134.58 ± 2.21); (134.24 ± 2.64); (134.75 ± 2.51); (134.64 ± 1.98); and (135.34 ±
4.57) pg/mL. Respectively. Moreover, their post-test concentration were(133,15 ±
4,01); (130,28 ± 3,59); (127,20 ± 5,56); (113,87 ± 4,30); and (120,87 ± 7,89)
pg/mL,respectively. Thus, the suplementation of CRLE to rats suffering from
hypercholesterolemia, seperti juga terlihat pada Tabel 2 bahwa terjadi penurunan
kadar IL-6 darah tikus Wistar Hypercholesterolemia.
Table 2
Decrease in IL-6 concentration in rats suffering from hypercholesterolemia
following supplementation of CRLE
Treatment
EDCR 0% (Kontrol)
EDCR 10 %
EDCR 15 %
EDCR 20 %
EDCR 25 %
Observation IL-6(pg/mL)
Pre-test
Pos-test
134,58 ± 2,21
133,15 ± 4,01
134,24 ± 2,64
130,28 ± 3,59
134,75 ± 2,51
127,20 ± 5,56
134,64 ± 1,98
113,87 ± 4,30
135,34 ± 4,57
120,87 ± 7,89
Extract leaf of Cantharanthus roseus contains flavonoids in abundance that have
anti-inflammatory property. When endothelium cells undergo inflammatory
activation, there are increase in expression of selectin, VCAM-1 and ICAM-1 that
promote adhesion of monocyte. Expression for molecule adhesion is induced by
pro-inflammatory cytokines such as IL-1Band TNF-α ,by acute-phase protein
CRP (produced in the liver as a response to IL-6), by protease activated receptor
signaling, by uptake of oxLDL through oxLDL receptor-1 (LOX-1), and by
interaction of CD40/CD40 ligands in tunica intima of the arteries (Bonetti et al.,
2003).
Interleukin-6 plays important role in inflammation. This cytokine is
released by activated macrophage, causing febrile so that it is called endogenous
pirogen. Cytokine IL-6 also initiates process related with acute response. Acute
phase of response is characterized by production of acute-phase protein by
hepatocyte. As has been demonstrated, acute inflammation is generally followed
by systemic acute response (Coico et al., 2003).
Supplementation of Cantharanthus roseus leaf extract
7
Conclusion
Supplementation of Cantharanthus roseus leaf extract (CRLE) at 20%
may significantly reduce the concentration of TNF-α in rats suffering from
hypercholesterolemia that can be related with inflammation condition. As TNF-α
is considered as a marker for the occurrence of inflammation, CRLE may be
regarded as having anti-inflammatory property. Likewise, such dosage of CRLE
(at 20%) may be used to reduce the concentration of cytokine interleukin-6 (IL-6)
which is also considered as a marker for inflammation.
References
[1] Anonim, Tapak Dara. Wikipedia Bahasa Indonesia. Indonesia: Wikimedia
Foundation Inc, 2009.
[2] Baraas - Faisa, kardiologi Molekuler Radikal Bebas, Disfungsi
EndotelAterosklerosis, Antioksidan.Latihan Fisik dan Rehabilitasi Jantung.
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Aterisoscler Thromb Vase Biol 23, (2003), 168-175.
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[5] Coico, R., S.. J. and Benyamin, E., Element of Innate and Aquiered Immunity
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Supplementation of Cantharanthus roseus leaf extract
9
[21]Wahjuni, S, Asupan Minyak Ikan Lemuru (Sardinella longiceps) Melalui
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Received: November 6, 2014; Published: December 11, 2014
HIKARI Ltd, www.m-hikari.com
http://dx.doi.org/10.12988/pacs.2015.41011
Supplementation of Cantharanthus Roseus Leaf
Extract as Anti-Inflammatory
Substance to Hypercholesterolemic Wistar
Derived-Rats
Sri Wahjuni1, Sri Rahayu Santi1 and A. A Ngurah Gunawan2
1
Department of Chemistry
Faculty of Mathematics and Natural Science
Udayana University, Bali, Indonesia
1
Department of Chemistry
Faculty of Mathematics and Natural Science
Udayana University, Bali, Indonesia
2
Department of Physics
Faculty of Mathematics and Natural Science
Udayana University, Bali, Indonesia
Copyright © 2014 Sri Wahjuni, Sri Rahayu Santi and A. A Ngurah Gunawan. This is an open
access article distributed under the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited.
Abstract
The present study on supplementation of Cantharanthus roseus leaf extract
(CRLE) to rats suffer from hypercholesterolemia was aimed to determine whether
it has property as an anti-inflammatory substance assessed from concentrations of
tumor necrosis factor-α (TNF- α) and Interleukin-6 (IL-6) in the blood serum. A
randomized pre-test and post-test design was employed, using 4 weeks old Wistar
derived-rats at body weight ranging from 70 – 72 gram. The rats were initially fed
with normal rat pellets for 1 month followed by high fat ration for 6 weeks in
order to induce hypercholesterolemia condition. Then they were divided into 5
groups according to various treatments applied as follows. Group 1: control –
2
Sri Wahjuni et al.
supplemented with 0% CRLE; Group 2: 10% CRLE; Group 3: 15% CRLE; Group
4: 20% CRLE; and Group 5: 25% CRLE. CRLE supplementation was last for 1
month. TNF- αconcentration was determined using Sandwich Enzyme (ELISA)
kit spectrophotometer (at π =500 nm) andconcentration of IL-6 was determined
using Quintikine HS immunoassay kit. In general, results of the current study
showed that CRLE may reduce inflammation based on assessment of
concentrations of TNF- α and IL-6 in the serum. Following supplementation of
20% CRLE, TNF- α concentration decreased from 28.62 pg/ml to 15.56 pg/ml
while IL-6 decreased from 134.64 g/ml to 113.87 pg/ml. In Indonesia,
Cantharanthus roseus grown as ornament plant and contain tannin, tripernoids,
and alkaloids. The component that acts an anti-inflammation is flavanoids.
Keywords: Leaf of Cantharanthus roseus, tumor necrosis factor (TNF),
interleukin-6 (IL-6), anti-inflammation, ELISA
Introduction
In Indonesia, research work on traditional medicine is still preferable for
combating animal diseases and human diseases. One of herbal plants that have
been proved beneficial for curing various diseases is Cantharanthus roseus (C.
roseus). It grown as ornament plants and its local name is “tapak dara ”. As
reported from many countries such as India and China, extract of C. roseus leaf
can be used to cure many diseases (Singh and Singh, 2001). It has been
discovered that extract leaf of C. roseus contains tannin, trypernoid, alkaloid, and
flavonoids. Flavonoids occur in the form of glycoside and it has been shown in
vivo that flavonoids may inhibit activity of NF-kB (nuclear factor kappa-B),
leading to inhibition of inflammation reaction (Han et al., 2007).
Inflammation is a complex condition that starts from tissue damaging.
Damage of the tissues are due to endogen factors (such as tissue necrosis) and to
external factors such as contact with foreign particles or infections. Inflammation
is generally a kind of response of vascular living tissues, that followed by
important endothelium process. Process occurring in the endothelium of blood
vessel lead in turn to development of atherosclerosis (Hosaya et al.,
2005).Atherosclerosis may become the consequence of hypercholesterolemia, also
called dyslipidemia – the increase in lipid content in the serum. Low density
lipoprotein (LDL) particularly the oxidized form may significantly cause damage
of the endothelium. On the other hand, high density lipoprotein (HDL) is the
protective component (Robbin and Cataren, 2002). Furthermore, chronic
hypercholesterolemia may result in cardiovascular diseases.
Prior to the occurrence of atherosclerosis and the subsequent diseases,
inflammation occurs. Thus, attempts must be made to reduce or minimized
inflammation reaction in order to avoid or minimize the effects of such various
health problems. For such aims, the current research work is designed to study the
Supplementation of Cantharanthus roseus leaf extract
3
effect of supplementation of extract leaf of herbal plant C. roseus as antiinflammatory substance, based on determination of pro-inflammatory cytokines
(TNF- α and IL-6) in the serum of experimental rats that underwent
hypercholesterolemia.
Materials and Methods
Materials
Animals used in the present study are: 50 male Wistar derived-rats at 4
weeks of age and weighing 70 – 72 grams. Experimental materials include C.
roseus leaf extract (CRLE), Sandwich enzyme immunoassay (ELISA) kit for
determination of TNF- α, Quintikine HS immunoassay kit for IL-6 determination,
EDTA, and diethyl ether. Serum collected from the experimental animals were
kept at -200C until assays were conducted. Prior to assays, serum samples were
centrifuged at 2000 rpmfor 10 minutes.
Methods
Experimental design employed was a randomized pre-test and post-test.
The experimental animals were divided into 5 groups according to the treatment
applied, namely Group 1 (control): supplemented with 0% CRLE; Group 2: 10%
CRLE; Group 3: 15% CRLE; Group 4: 20% CRLE; and Group 5: 25%
CRLE.Prior to supplementation of CRLE, all experimental animals were initially
raised under normal rat pellets for 1 month. Then, they are all subjected to high fat
ration for 6 weekin order to make them suffering from hypercholesterolemia.Such
feeding treatment has been shown to result in hypercholesterolemia and chronic
inflamationin rats (Wahjuni,2011). At the end of the high fat feeding phase,
concentrations of TNF-α and IL-6 were measured and the values were considered
as the pre-test data. Supplementation of the various dosages of CRLE to the
experimental animals lasted for 6 weeks. At the end of this phase, concentrations
of TNF-α and IL-6 were again measured and the data was considered as the posttest values
The data was analyzed descriptively. Data selection including editing,
coding and tabulation was performed using Windows file navigator statistical
program (Triton, 2006; Pramesti, 2007). For normality analysis of TNF-α and IL6 data, Shapiro Wilk test was employed at 5% level of significant (p = 0.05). H0
hypothesis: frequency of observation/ frequency of expectation. Based on this
test, the data was considered to be normally distributed when P > 0.05. Moreover,
variant homogeneity was analyzed using the Levent’s test.Ho hypothesis:
variation of control group = variation of Group 1 = variation of Group 2 =
variation of Group 3 = variation of Group 4. Ho was accepted when P > a, and
vice versa .
4
Sri Wahjuni et al.
Result and discussion
Decrease in TNF-α concentration
The average level of TNF-α measured from blood of 50 male rats suffering
from hypercholesterolemia at pre-test and post-test was presented at Table 1. The
pre-test TNF-α values for control group, Group 1, Group 2, Group 3 and Group 4
were (28.98 ± 6.00); (29.12 ± 5.79);(29.02 ± 5.34); (28.62 ± 4.72); dan (29,02 ±
5.06) pg/mLrespectively. Meanwhile, the post-test values for control group,
Group 1, Group 2, Group 3 and Group 4 were (28.11 ± 5.94); (27.32
±5.01);(24.42 ± 5.74); (15,56 ±7,20); ( 26,02 ± 8,34 )pg/mL respectively. When
the pre-test and post-test values were compared, there was no significant reduction
in concentration of TNF-α for the control group. On the other hand,
supplementation of CRLE at 10%, 15%, 20% and 25% has resulted in decreasing
level of TNF-α. Supplementation of CRLE at 10% and 15% led to reduction of
TNF-α level up to 0.79 and 3.69 pg/ml, respectively. However, statistical analysis
showed that such differences were not significant (P> 0.05). But a significant
difference (P < 0.05) in TNF-α concentrations between the pre-test and post-test
values was noted for rats subjected to supplementation of 20% CRLE (Group 3).
For this group, TNF-α concentration has decreased at amount of 12.55 pg/ml.
Furthermore, for rats under 25% CRLE supplementation, no significant different
(P > 0.05) in TNF-α concentration was observed.
Tablel 1
Decrease level of TNF-α serum Wistar Rat Hiperkolesterolemia
Treatment
Observation TNF-α (pg /mL)
Pre-test
Pos-test
EDGR 0 % (control)
EDGR 10%
EDGR 15%
EDGR 20%
EDGR 25%
28.98 ± 6.00
29,12± 5,79
29,02 ± 5,34
28,62 ± 4,72
29,02 ± 5.06
28.11 ± 5.94
27,32 ± 5,01
24,42 ± 5,74
15,56 ± 7,20
26,02 ± 8,34
The present study showed that supplementation of 20% CRLE has resulted
in the highest reduction in the serum TNF-α concentration. But supplementation
of CRLE at 25% did not lead to further decrease in TNF-α concentration. It seems
that at 25% supplementation, the response has reached saturated state, so that
cannot cause further decreasing effect on TNF-α concentration. The present result
on supplementation of 25% CRLE seems in accordance with the findings of Chen
and Goeddel (2002) who reported that TNF-αproduced by NF-KB do not undergo
transcription reaction when it reaches saturated state. Therefore, transcription
factors are responsible for induction of gene expression for inflammation process
to occur.
Supplementation of Cantharanthus roseus leaf extract
5
Inflammation is generally a response to external and internal factors by
vascular living tissues. The inflammation response is usually followed by an
important process – endothelium process. Endothelium of blood vessel is the site
where the deposition of fat occurs and known as atherosclerosis. Endothelium is
the primary target for mechanical and chemical injuries caused by
hypercholesterolemia. Chronic hypercholesterolemia has been known to result in
the shift of acute pro-inflammation and prothrombosis responses into the chronic
responses. These responses, in turn, are followed by infiltration of leucocytes,
particularly of monocyte, to sub-endothelium, and subsequentlychanging into
macrophage cells. The macrophage cells then phagocyte the remnant of LDL
which is oxidized to form atheroma (Baraas, 2006).
Simopolous (2002) reported that flavonoids as anti-inflammation play role
as immunomodulator. Mechanismeis increas antioxtidan endogen and capcing
free radikal.Inhibition stres oksidatif by some flavonoid incontain of
cantharanthus roseus Leaf Extract (Figure 1). And than flavonoid cantharanthus
roseus isolasitian metabolit sekunder compound, extraction, distilation,
purification, dan identificationby GC-MS(Gas Chromatograph Mass
Spectrometer ) Shimadzu QP2010, identifay flavonoid what typeCantharanthus
roseus Leaf Extract.
Figure 1: Habition Flavonoid 0f NF-kB (Nuclear Factor Kappa Beta)was decreas
habition
interleukin product pro inflamation, COX2 can habition reaction
inflamation.
Its role as anti-inflammation is due to the weakening effect of arachidonic acid.
Arachidonic acid may activate cyclooxygenase and 5-lipooxygenase enzymes.
These two enzymes act as endothelium-dependent vasodilator that cause
relaxation of normal coronary arteries and cause paradox vasoconstriction of
arteries in hypercholesterolemia condition.
6
Sri Wahjuni et al.
Decrease in IL-6 concentration
Concentrations of interleukin-6 (IL-6) in blood serum of rats suffering
from hypercholesterolemia and then treated with CRLE at pre-test and post-test
are presented in Table 2. The pre-test IL-6 concentrations for group supplemented
with 0% CRLE (control), with 10%, 15%, 20%, and with 25% CRLE were
(134.58 ± 2.21); (134.24 ± 2.64); (134.75 ± 2.51); (134.64 ± 1.98); and (135.34 ±
4.57) pg/mL. Respectively. Moreover, their post-test concentration were(133,15 ±
4,01); (130,28 ± 3,59); (127,20 ± 5,56); (113,87 ± 4,30); and (120,87 ± 7,89)
pg/mL,respectively. Thus, the suplementation of CRLE to rats suffering from
hypercholesterolemia, seperti juga terlihat pada Tabel 2 bahwa terjadi penurunan
kadar IL-6 darah tikus Wistar Hypercholesterolemia.
Table 2
Decrease in IL-6 concentration in rats suffering from hypercholesterolemia
following supplementation of CRLE
Treatment
EDCR 0% (Kontrol)
EDCR 10 %
EDCR 15 %
EDCR 20 %
EDCR 25 %
Observation IL-6(pg/mL)
Pre-test
Pos-test
134,58 ± 2,21
133,15 ± 4,01
134,24 ± 2,64
130,28 ± 3,59
134,75 ± 2,51
127,20 ± 5,56
134,64 ± 1,98
113,87 ± 4,30
135,34 ± 4,57
120,87 ± 7,89
Extract leaf of Cantharanthus roseus contains flavonoids in abundance that have
anti-inflammatory property. When endothelium cells undergo inflammatory
activation, there are increase in expression of selectin, VCAM-1 and ICAM-1 that
promote adhesion of monocyte. Expression for molecule adhesion is induced by
pro-inflammatory cytokines such as IL-1Band TNF-α ,by acute-phase protein
CRP (produced in the liver as a response to IL-6), by protease activated receptor
signaling, by uptake of oxLDL through oxLDL receptor-1 (LOX-1), and by
interaction of CD40/CD40 ligands in tunica intima of the arteries (Bonetti et al.,
2003).
Interleukin-6 plays important role in inflammation. This cytokine is
released by activated macrophage, causing febrile so that it is called endogenous
pirogen. Cytokine IL-6 also initiates process related with acute response. Acute
phase of response is characterized by production of acute-phase protein by
hepatocyte. As has been demonstrated, acute inflammation is generally followed
by systemic acute response (Coico et al., 2003).
Supplementation of Cantharanthus roseus leaf extract
7
Conclusion
Supplementation of Cantharanthus roseus leaf extract (CRLE) at 20%
may significantly reduce the concentration of TNF-α in rats suffering from
hypercholesterolemia that can be related with inflammation condition. As TNF-α
is considered as a marker for the occurrence of inflammation, CRLE may be
regarded as having anti-inflammatory property. Likewise, such dosage of CRLE
(at 20%) may be used to reduce the concentration of cytokine interleukin-6 (IL-6)
which is also considered as a marker for inflammation.
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Received: November 6, 2014; Published: December 11, 2014