The Potential of Total Flavonoids and Phenol Contents in Stem Bark of Gayam (Inocarpus fagiferus Fosb) as an Antioxidant through the Decrease of MDA Level, Increase of SOD Activities and Improvement o.
ejbps, 2015, Volume 2, Issue 6, 55-58.
Rahayu et al.
SJIF Impact Factor 2.062
Research Article
ISSN 2349-8870
European of
Journal
of Biomedical and Pharmaceutical Sciences
European Journal
Biomedical
Volume: 2
Issue: 6
AND Pharmaceutical sciences
55-58
Year: 2015
http://www.ejbps.com
THE POTENTIAL OF TOTAL FLAVONOIDS AND PHENOL CONTENTS IN STEM
BARK OF GAYAM (Inocarpus fagiferus Fosb) AS AN ANTIOXIDANT THROUGH THE
DECREASE OF MDA LEVEL, INCREASE OF SOD ACTIVITIES AND IMPROVEMENT
OF LIPID PROFILE IN RATS ATHEROSCLEROSIS HYPERCHOLESTEROLEMIA
*Sri Rahayu Santi, I Made Sukadana and I Made Siaka
Department of Chemistry-Faculty of Science and Maths
*Author for Correspondence: Sri Rahayu Santi
Department of Chemistry-Faculty of Science and Maths
Article Received on 10/09/2015
Article Revised on 30/09/2015
Article Accepted on 22/10/2015
ABSTRACT
This study aimed to demonstrate the antioxidant potential of n-buthanol extract of gayam stem bark in
preventing atherosclerosis through the decrease of malondialdehyde (MDA) level, lipid profile improvements
such as the decrease of total cholesterol, triglycerides, LDL cholesterol (LDL-cholest) and the increase of
HDL cholesterol (HDL-cholest) levels, and SOD activity in Wistar rats that were treated with a high fat
diet or hypercholesterolemia conditions within 3 months of observation. The study was initiated by preparing
n-buthanol extract of gayam stem bark which was obtained from the partition of 95 g ethanol extract of gayam
stem bark. A total of 30 g of n-buthanol concentrate extract was obtained and then was applied to Wistar rats
for 3 months with the use of the posttest only control group design. Twenty-five Wistar rats were randomized
into 5 groups, K1 (negative control), K2 (positive control, hypercholesterolemia), P1 (hypercholesterolemia +
n-buthanol extract in the dose of 50 mg/kg bw), P2 (hypercholesterolemia + n-buthanol extract in the dose of
100 mg/kg bw), and P3 (hypercholesterolemia + n-buthanol extract in the dose of 150 mg/kg bw). The results
showed that n-buthanol extract of gayam stem bark in the dose of 50 mg/kg bw was able to decrease the levels
of MDA, total cholesterol, triglycerides, and increase the SOD activity, while in the dose of 100 mg/kg bw the
extract could decrease the LDL cholesterol and increase of HDL cholesterol levels of blood plasm in wistar
rats hypercholesterolemia.
KEYWORDS: Inocarpus fagiferus Fosb, Atherosclerosis, MDA, lipid profile, SOD activity.
I. INTRODUCTION
Medicinal plants are one source of natural
antioxidants that can increase endogenous antioxidant
and hence, it may reduce the risk of certain diseases such
as cancer, heart disease, neurodegenerative diseases,
stroke,
inflammation,
and
atherosclerosis.[1,2]
Secondary metabolites such as natural phenols,
flavonoids, and their derivatives contained in medicinal
plants are potentially used as a source of natural
antioxidants for free radicals scavenger.[2,3] Gayam
(Inocarpus Fagiferus Fosb) or in Bali known as gatep
is one of the herbs that empirically in the Fiji Islands
is used as a drug to prevent heart disease and
atherosclerosis.[4] The stem bark of gayam contains
compounds such as triterpenoids, anthraquinone,
steroids and flavonoids and phenols as the main
content. n-buthanol extract of gayam stem bark
contains total flavonoids and phenols respectively by
0.09% and 14.16%, that are able to capture the free
radicals DPPH by 50% at a concentration of 20 ppm,
and are able to inhibit the formation of lipid peroxide
www.ejbps.com
by 63.03%. The antioxidant activity of n-buthanol
extract of gayam stem bark in vitro against DPPH is
greater than the antioxidant activity of vitamin E which
provides IC50 at a concentration of 25 ppm, as
well as its ability to inhibit the formation of lipid
peroxides by 50.39 %.[5] Therefore, the potential of
antioxidant activity of n-buthanol extract of gayam
stem bark is required to be further investigated in
vivo by determining the activity of SOD, levels of
malondialdehyde (MDA), as well as its effect on lipid
profile improvements such as total cholesterol,
triglycerides, LDL cholesterol (LDL-cholest) and
HDL cholesterol (HDL-cholest) as a marker to
prevent the occurrence of atherosclerosis in wistar rats
with high fat diet or hypercholesterolemia conditions.[6,7]
This study discusses the differences in the
levels of MDA, total cholesterol, triglycerides,
HDL cholesterol, LDL cholesterol, and SOD activity
of each treatment group compared with the control
group of hypercholesterolemia, as the marker of cell
55
Rahayu et al.
European Journal of Biomedical and Pharmaceutical Sciences
membrane damage due to lipid peroxidation, [8,9,10] an
indirect marker of fatty acid oxidation products,[11] and
the marker if n-buthanol extract of gayam stem bark
could be SOD inducer to the endogenous antioxidant.
mg/kg bw
P3
= group of wistar rats fed with a high-fat diet
+ n-buthanol extract of gayam stem bark in the dose of
150 mg/kg bw
II. MATERIALS AND METHODS
The Research Materials
The plant material was obtained from Tabanan Bali
and its classification has been determined. Chemicals
used in this study were ethanol (technical and pa),
chloroform (technical and pa), ethyl acetate (technical
and pa), n-buthanol, and blood plasm of wistar rat.
Chemicals required for plasm analysis were
TBARS
(R&D
Systems,
Cat.
ParameterTM
KGE013), Superoxide Dismutase (SOD) Activity
Assay Kit (BioVision, Cat. K335-100). The
determination of profile lipid was conducted at UPT.
Balai Laboratorium Kesehatan Provinsi Bali. All of the
rats were treated in accordance with the rules of the
etical clerence of Ethics Committee of Animals Use in
Research and Education of the Faculty of Veterinary
Udayana University.
After 12 weeks, the blood plasm of all of the rats such
as the control groups (K1 and K2) and the treatment
groups (P1, P2, and P3) were drawn for MDA, SOD
activity, total cholesterol, triglycerides, HDL-cholest
and LDL-cholest analyses. The difference of all
variables were analyzed by one way Anova with
=
0.05.
III. RESULTS AND DISCUSSION
3.1 MDA Blood Plasm
The results of MDA levels of blood plasm
all the rats (K1, K2, P1, P2, and P3)
significant decreases in the level of MDA
treated with n-buthanol extract in the doses
and 150 mg/kg bw as described in Figure 1.
analysis of
showed a
in the rats
of 50, 100,
Research Instruments
The equipment used in this study was a set of glasses,
blender, sieve, extractor, rotary vacuum evaporator,
centrifuges,
analytical
balance,
thermometer,
micropipette, pipette, volumetric flask, stomach probes,
syringes, and UV-vis spectrophotometer.
Researchers procedure
a. Preparation of n-buthanol extract of gayam stem
bark
As much as 3 kg dried powder of gayam stem bark
was macerated with ethanol for 24 hours. The extract
was filtered, while the crude was remacerated several
times with ethanol until the compound contained in
the sample was extracted out. The filtrate obtained
was evaporated with a rotary vacuum evaporator to
obtain a concentrate extract of ethanol. The concentrate
ethanol extract was further diluted with ethanol-water
mixture (7: 3) and then partitioned successively with
chloroform, ethyl acetate and n-buthanol. The extract
obtained was evaporated and weighed. n-buthanol
extract was then applied to the rats.
b. Applications of n-buthanol extract of gayam stem
bark on Wistar Rats
n-buthanol extract was applied at various doses i.e 50
mg/kg bw, 100 mg/kg bw, and 150 mg/kg bw. Twenty
five wistar rats were randomized into 5 groups with
posttest only control group design.[12] as follows:
K1
= group of wistar rats fed with standard diet
(negative control) K2
=
group of wistar rats
fed with a high-fat diet (positive control)
P1
= group of wistar rats fed with a high-fat diet
+ n-buthanol extract of gayam stem bark in the dose of
50 mg/kg bw
P2
= group of wistar rats fed a high-fat diet + nbuthanol extract of gayam stem bark in the dose of 100
www.ejbps.com
Figure 1.
The average of MDA levels of K1 group, treatment
groups (P1, P2, and P3) toward K2. The difference mean
of groups were: K1 vs K2, p
Rahayu et al.
SJIF Impact Factor 2.062
Research Article
ISSN 2349-8870
European of
Journal
of Biomedical and Pharmaceutical Sciences
European Journal
Biomedical
Volume: 2
Issue: 6
AND Pharmaceutical sciences
55-58
Year: 2015
http://www.ejbps.com
THE POTENTIAL OF TOTAL FLAVONOIDS AND PHENOL CONTENTS IN STEM
BARK OF GAYAM (Inocarpus fagiferus Fosb) AS AN ANTIOXIDANT THROUGH THE
DECREASE OF MDA LEVEL, INCREASE OF SOD ACTIVITIES AND IMPROVEMENT
OF LIPID PROFILE IN RATS ATHEROSCLEROSIS HYPERCHOLESTEROLEMIA
*Sri Rahayu Santi, I Made Sukadana and I Made Siaka
Department of Chemistry-Faculty of Science and Maths
*Author for Correspondence: Sri Rahayu Santi
Department of Chemistry-Faculty of Science and Maths
Article Received on 10/09/2015
Article Revised on 30/09/2015
Article Accepted on 22/10/2015
ABSTRACT
This study aimed to demonstrate the antioxidant potential of n-buthanol extract of gayam stem bark in
preventing atherosclerosis through the decrease of malondialdehyde (MDA) level, lipid profile improvements
such as the decrease of total cholesterol, triglycerides, LDL cholesterol (LDL-cholest) and the increase of
HDL cholesterol (HDL-cholest) levels, and SOD activity in Wistar rats that were treated with a high fat
diet or hypercholesterolemia conditions within 3 months of observation. The study was initiated by preparing
n-buthanol extract of gayam stem bark which was obtained from the partition of 95 g ethanol extract of gayam
stem bark. A total of 30 g of n-buthanol concentrate extract was obtained and then was applied to Wistar rats
for 3 months with the use of the posttest only control group design. Twenty-five Wistar rats were randomized
into 5 groups, K1 (negative control), K2 (positive control, hypercholesterolemia), P1 (hypercholesterolemia +
n-buthanol extract in the dose of 50 mg/kg bw), P2 (hypercholesterolemia + n-buthanol extract in the dose of
100 mg/kg bw), and P3 (hypercholesterolemia + n-buthanol extract in the dose of 150 mg/kg bw). The results
showed that n-buthanol extract of gayam stem bark in the dose of 50 mg/kg bw was able to decrease the levels
of MDA, total cholesterol, triglycerides, and increase the SOD activity, while in the dose of 100 mg/kg bw the
extract could decrease the LDL cholesterol and increase of HDL cholesterol levels of blood plasm in wistar
rats hypercholesterolemia.
KEYWORDS: Inocarpus fagiferus Fosb, Atherosclerosis, MDA, lipid profile, SOD activity.
I. INTRODUCTION
Medicinal plants are one source of natural
antioxidants that can increase endogenous antioxidant
and hence, it may reduce the risk of certain diseases such
as cancer, heart disease, neurodegenerative diseases,
stroke,
inflammation,
and
atherosclerosis.[1,2]
Secondary metabolites such as natural phenols,
flavonoids, and their derivatives contained in medicinal
plants are potentially used as a source of natural
antioxidants for free radicals scavenger.[2,3] Gayam
(Inocarpus Fagiferus Fosb) or in Bali known as gatep
is one of the herbs that empirically in the Fiji Islands
is used as a drug to prevent heart disease and
atherosclerosis.[4] The stem bark of gayam contains
compounds such as triterpenoids, anthraquinone,
steroids and flavonoids and phenols as the main
content. n-buthanol extract of gayam stem bark
contains total flavonoids and phenols respectively by
0.09% and 14.16%, that are able to capture the free
radicals DPPH by 50% at a concentration of 20 ppm,
and are able to inhibit the formation of lipid peroxide
www.ejbps.com
by 63.03%. The antioxidant activity of n-buthanol
extract of gayam stem bark in vitro against DPPH is
greater than the antioxidant activity of vitamin E which
provides IC50 at a concentration of 25 ppm, as
well as its ability to inhibit the formation of lipid
peroxides by 50.39 %.[5] Therefore, the potential of
antioxidant activity of n-buthanol extract of gayam
stem bark is required to be further investigated in
vivo by determining the activity of SOD, levels of
malondialdehyde (MDA), as well as its effect on lipid
profile improvements such as total cholesterol,
triglycerides, LDL cholesterol (LDL-cholest) and
HDL cholesterol (HDL-cholest) as a marker to
prevent the occurrence of atherosclerosis in wistar rats
with high fat diet or hypercholesterolemia conditions.[6,7]
This study discusses the differences in the
levels of MDA, total cholesterol, triglycerides,
HDL cholesterol, LDL cholesterol, and SOD activity
of each treatment group compared with the control
group of hypercholesterolemia, as the marker of cell
55
Rahayu et al.
European Journal of Biomedical and Pharmaceutical Sciences
membrane damage due to lipid peroxidation, [8,9,10] an
indirect marker of fatty acid oxidation products,[11] and
the marker if n-buthanol extract of gayam stem bark
could be SOD inducer to the endogenous antioxidant.
mg/kg bw
P3
= group of wistar rats fed with a high-fat diet
+ n-buthanol extract of gayam stem bark in the dose of
150 mg/kg bw
II. MATERIALS AND METHODS
The Research Materials
The plant material was obtained from Tabanan Bali
and its classification has been determined. Chemicals
used in this study were ethanol (technical and pa),
chloroform (technical and pa), ethyl acetate (technical
and pa), n-buthanol, and blood plasm of wistar rat.
Chemicals required for plasm analysis were
TBARS
(R&D
Systems,
Cat.
ParameterTM
KGE013), Superoxide Dismutase (SOD) Activity
Assay Kit (BioVision, Cat. K335-100). The
determination of profile lipid was conducted at UPT.
Balai Laboratorium Kesehatan Provinsi Bali. All of the
rats were treated in accordance with the rules of the
etical clerence of Ethics Committee of Animals Use in
Research and Education of the Faculty of Veterinary
Udayana University.
After 12 weeks, the blood plasm of all of the rats such
as the control groups (K1 and K2) and the treatment
groups (P1, P2, and P3) were drawn for MDA, SOD
activity, total cholesterol, triglycerides, HDL-cholest
and LDL-cholest analyses. The difference of all
variables were analyzed by one way Anova with
=
0.05.
III. RESULTS AND DISCUSSION
3.1 MDA Blood Plasm
The results of MDA levels of blood plasm
all the rats (K1, K2, P1, P2, and P3)
significant decreases in the level of MDA
treated with n-buthanol extract in the doses
and 150 mg/kg bw as described in Figure 1.
analysis of
showed a
in the rats
of 50, 100,
Research Instruments
The equipment used in this study was a set of glasses,
blender, sieve, extractor, rotary vacuum evaporator,
centrifuges,
analytical
balance,
thermometer,
micropipette, pipette, volumetric flask, stomach probes,
syringes, and UV-vis spectrophotometer.
Researchers procedure
a. Preparation of n-buthanol extract of gayam stem
bark
As much as 3 kg dried powder of gayam stem bark
was macerated with ethanol for 24 hours. The extract
was filtered, while the crude was remacerated several
times with ethanol until the compound contained in
the sample was extracted out. The filtrate obtained
was evaporated with a rotary vacuum evaporator to
obtain a concentrate extract of ethanol. The concentrate
ethanol extract was further diluted with ethanol-water
mixture (7: 3) and then partitioned successively with
chloroform, ethyl acetate and n-buthanol. The extract
obtained was evaporated and weighed. n-buthanol
extract was then applied to the rats.
b. Applications of n-buthanol extract of gayam stem
bark on Wistar Rats
n-buthanol extract was applied at various doses i.e 50
mg/kg bw, 100 mg/kg bw, and 150 mg/kg bw. Twenty
five wistar rats were randomized into 5 groups with
posttest only control group design.[12] as follows:
K1
= group of wistar rats fed with standard diet
(negative control) K2
=
group of wistar rats
fed with a high-fat diet (positive control)
P1
= group of wistar rats fed with a high-fat diet
+ n-buthanol extract of gayam stem bark in the dose of
50 mg/kg bw
P2
= group of wistar rats fed a high-fat diet + nbuthanol extract of gayam stem bark in the dose of 100
www.ejbps.com
Figure 1.
The average of MDA levels of K1 group, treatment
groups (P1, P2, and P3) toward K2. The difference mean
of groups were: K1 vs K2, p