Honey improves lipid profile of diet-induced hypercholesterolemic rats

177

*Department of Physiology,
Faculty of Medicine,
Islamic University of Indonesia

Correspondence:
dr. Titis Nurmasitoh, M.Sc.
Department of Physiology,
Faculty of Medicine,
Islamic University of Indonesia,
Jln. Kaliurang km 14,5
Yogyakarta 55584
Phone: +62815 685 7437
Email: titisnurmasitoh@gmail.com

Univ Med 2015;34:177-86
DOI: 10.18051/UnivMed.2016.v35.177-186
pISSN: 1907-3062 / eISSN: 2407-2230

This open access article is distributed under
a Creative Commons Attribution-Non
Commercial-Share Alike 4.0 International
License

ABSTRACT

UNIVERSA MEDICINA
September-December, 2015September-December, 2015September-December, 2015September-December, 2015September-December, 2015 Vol.34 - No.3 Vol.34 - No.3 Vol.34 - No.3 Vol.34 - No.3 Vol.34 - No.3

BACKGROUND
Coronary heart disease (CHD) is a major cause of morbidity and mortality
throughout the world, including Indonesia. One of the risk factors for
CHD is hypercholesterolemia. One of the natural products that has been
developed for the treatment of hypercholesterolemia is honey. Honey
contains fructooligosaccharides, various vitamins, minerals, and enzymes
which are supposedly able to lower blood cholesterol levels. This research
aimed to study the influence of honey on the levels of blood total
cholesterol, triglyceride, and low density lipoprotein (LDL) levels in Wistar
rats.

METHODS
This study was of experimental post test control group design. Twenty-
four male Wistar rats (Rattus norvegicus) were randomly divided into 4
groups. K1 was the negative control group (with normal diet), K2 was
the positive control group (with high-fat diet), P1 was fed a high-fat diet
for 7 days, followed by high-fat diet plus honey for the next 7 days. P2
was fed a high-fat diet for 7 days, followed by regular diet plus honey for
the next 7 days. After completion of this treatment, total cholesterol,
triglycerides, and LDL levels were measured by the cholesterol oxidase
phenol+aminophenazone (CHOD-PAP) method using enzymatic
spectrophotometry principles.

RESULTS
There were significant differences in total cholesterol, triglyceride, and
LDL levels between all groups after day 15 (p<0.05).

CONCLUSION
Honey supplementation was able to reduce the blood levels of total
cholesterol, triglycerides, and LDL. Honey supplementation accompanied
by non-cholesterol feeds could more effectively lower total cholesterol,
triglycerides, and LDL serum levels in Wistar rats.

Keywords: Cholesterol, triglycerides, LDL, honey, hypercholesterolemia,
rats

Honey improves lipid profile of diet-induced
hypercholesterolemic rats

Titis Nurmasitoh* and Miranti Dewi Pramaningtyas*

DOI: http://dx.doi.org/10.18051/UnivMed.2015.v34.177-186

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Nurmasitoh, Pramaningtyas Honey improves hypercholesterolemic

Madu memperbaiki profil lipid pada tikus
yang diinduksi diet hiperkolesterolemia

LATAR BELAKANG
Penyakit jantung koroner (PJK) merupakan penyebab utama kesakitan dan kematian di seluruh dunia, termasuk
Indonesia. Salah satu faktor risiko PJK adalah hiperkolesterolemia. Salah satu bahan alami yang banyak
dikembangkan dalam pengobatan adalah madu. Madu mengandung fruktooligoakarida, berbagai vitamin, mineral,
dan enzim yang diduga dapat menurunkan kadar kolesterol dalam darah. Penelitian ini bertujuan untuk mempelajari
pengaruh madu terhadap kadar kolesterol total, trigliserida, dan LDL darah.

METODE
Penelitian ini merupakan penelitian eksperimental menggunakan post test control group design. Dua puluh empat
ekor tikus Wistar (Rattus norvegicus) jantan dibagi secara acak dalam 4 kelompok. Kelompok K1 adalah kontrol
negatif (diberi pakan biasa), K2 adalah kontrol positif (diberi pakan tinggi lemak), P1adalah kelompok yang
diberi pakan tinggi lemak selama 7 hari dilanjutkan pakan tinggi lemak ditambah madu selama 7 hari berikutnya,
dan P2 adalah kelompok yang diberi pakan tinggi lemak selama 7 hari dilanjutkan pakan biasa ditambah madu
selama 7 hari berikutnya. Setelah selesai perlakuan, darah diambil untuk diperiksa kadar kolesterol total, trigliserid,
dan LDL menggunakan metode CHOD-PAP dengan prinsip spektrofotometri enzimatis.

HASIL
Terdapat perbedaan bermakna kadar kolesterol, trigliserida, dan LDL setelah hari ke-15 antara keempat kelompok
(p<0,05).

KESIMPULAN
Pemberian suplementasi madu dapat menurunkan kadar kolesterol total, trigliserida, dan LDL dalam darah pada
tikus. Pemberian suplementasi madu disertai dengan pemberian pakan non-kolesterol dapat menurunkan kadar
kolesterol total, trigliserida, dan LDL dalam darah secara lebih efektif.

Kata kunci: Kolesterol, trigliserida, LDL, madu, hiperkolesterolemia,tikus

ABSTRAK

INTRODUCTION

Coronary heart disease (CHD) is a major
cause of morbidity and mortality throughout the
world, including Indonesia. During the last fifty
years, the number of CHD patients has continued
t o i n c r e a s e , w i t h s u d d e n d e a t h a s f i n a l
outcome.(1-4) The increased incidence of CHD is
associated with increased CHD risk factors such
as dyslipidemia, obesity, hypertension, smoking
habit, diabetes mellitus, and other risk factors
related to lifestyle. In order to decrease CHD
cases, the risk factors should be controlled. As

the increased levels of blood cholesterol is also
a risk factor for CHD, a decrease in total
cholesterol levels, especially reduced levels of
low-density lipoprotein (LDL), is one of the
therapeutic targets to prevent and decrease the
incidence of CHD.(1,3,5)

The American College of Cardiology and the
American Heart Association (ACC/AHA) have
issued guidelines for cholesterol therapy (also
known as Cholesterol Treatment Guidelines,
CTG). The main treatment for
hypercholesterolemia is by the use of statins.
Statins act to lower blood cholesterol levels by

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Univ Med Vol. 34 No.3

inhibiting 3-hydroxy-3-methyl-glutaryl-coenzyme
A (HMG-CoA) reductase.(6) Several other
substances used to lower cholesterol, are among
others bile-acid sequestrants (inhibiting the
absorption of bile acids), vitamin E, and
gemfibrozil. So far, the use of statins is the most
effective strategy to lower blood LDL levels.(1)

Statins can lower blood cholesterol in more than
30% of patients with hypercholesterolemia.
However, the use of statins and other anti-
cholesterol drugs, especially in the long term,
might cause permanent side effects, such as those
associated with gastrointestinal symptoms, skin
rash, anxiety disorders, and hepatic
dysfunction.(7,8) Therefore, controlling risk factors
by improving lifestyle and performing early
monitoring is the best measure to prevent CHD
caused by hypercholesterolemia.

Various studies on the benefits of honey have
been performed. Honey is widely used empirically
for its anti-inflammatory, antibacterial, antifungal,
antiviral, antihypertensive, antioxidant,
cardioprotective, hypoglycemic and other
actions.(3,9,10) However, scientific evidence
supporting honey as a medicine still needs to be
developed to optimize its utilization. Various
literature reports mention that honey contains a
complete array of nutrients and has the potential
to be developed into a medication. Various
vitamins, minerals, fructo-oligosaccharides,
polyphenols, amino acids, and enzymes contained
in honey are expected to be useful for lowering
blood cholesterol levels.

This research aimed to study the influence
of honey on the levels of total cholesterol,
triglycerides, and LDL in the blood of diet-induced
hypercholesterolemic Wistar rats (Rattus
norvegicus).

METHODS

Research design
This was an experimental laboratory

research using post test control group design. The
study took place in the Physiology Laboratory,
Faculty of Medicine, Islamic University of

Indonesia. The study was conducted from January
to May 2010.

Animals
A total of twenty four male Wistar rats

(Rattus norvegicus) aged 15 weeks, weighing 200-
250 grams, were obtained from the animal house
at the Integrated Research and Testing Laboraory
(Laboratorium Penelitian dan Pengujian
Terpadu, LPPT), Gadjah Mada University,
Yogyakarta. The number of rats needed was
determined based on the 3Rs principle
(replacement, refinement, and reduction) and was
calculated using the following equation:

E = N-T
where: E = a constant between 10-20, N = planned
number of animals per group multiplied by
number of treatment groups, T = number of
treatment groups.

This study used 4 groups. If N or the number
of animals per group = 6, then E = (6x4) -4,
therefore E=20, which is still included in the
specified range of values between 10-20. Thus,
the total number of rats was 24 and each group
contained 6 rats.(11) Before the treatment was
performed, the rats were isolated for an adaptation
stage of 7 days. They were placed in cages with
regulated indoor light intensity, consisting of 12
hours light and 12 hours dark periods. During
this stage, feed and water were given ad libitum.

Treatment
Rats were randomly assigned into four

groups, namely the negative control group (K1),
the positive control group (K2), treatment group
1 (P1), and treatment group 2 (P2). Each group
of six rats was assigned randomly. Group K1 was
given regular feed from the beginning to the end
of the study. Group K2 was given a high-fat diet
for 14 days. Group P1 was given high-fat feeds
for 7 days, followed by high-fat feeds plus honey
for the next 7 days. Group P2 was given high-fat
feeds for 7 days, followed by regular feed plus
honey for the next 7 days. All rats were weighed
daily to determine changes in body weight from
day to day.

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Nurmasitoh, Pramaningtyas Honey improves hypercholesterolemic

Preparation of honey
Honey from one local honey store was

chosen to be used in the study. Based on the
printed label of the packaging, the honey used
for this study contained glucose, fructose,
sucrose, amino acids, zinc, anti-oxidant,
potassium, phosporus, iron, vitamins (B1, B2),
and some enzymes. The honey was given daily
by gavage at a dose of 0.8 mL/day. The dose of
honey for the rats was obtained by multiplying
the usual dose of honey for daily human
consumption by a conversion factor. The usual
dose for human consumption as printed on the
packaging and empirically considered adequate
to reduce cholesterol was 2 tablespoons (30 mL)
each day. After multiplying the dose for humans
by the conversion factor of man to rat (0.018), a
dose of 0.8 mL/day was obtained. Each dose of
honey was dissolved in water to a volume of 2
ml. The honey used in this experiment belonged
to the kapok tree type of honey, which had been
commercialized with a specific brand.

Preparation of high-fat diet
High-fat feed was prepared by mixing the

regular feed with white butter at a ratio of 5:1,
i.e. 5 parts of feed (20 g/day/rat) was mixed with
1 part white butter (4 grams/day). The dose of
white butter was obtained from a preliminary
study. White butter was chosen in this study on
the basis of economic and accessability aspects.
The feed and butter were evenly mixed and then
placed in a feed container in each cage, to be
consumed by the rats. The amount of feed
consumed was determined daily by measuring
the feed left in the container.

Biochemical analyses
Blood sampling of the rats was performed

three times, i.e. before treatment, on day 7 of
treatment, and on day 15, after completion of the
treatment. Blood was taken at the retro-orbital
plexus using 2 ml hematocrit pipettes. The blood
samples were taken to the Laboratory of Nutrition,
Central Inter-University Laboratory, Gadjah
Mada University, to check the levels of total

cholesterol, triglycerides, and low density
cholesterol (LDL) using the cholesterol oxidase
phenol + aminophenazone (CHOD-PAP) method,
based on the principles of enzymatic
spectrophotometry. The results were expressed in
mg/dL.

Statistical analysis
The data on total cholesterol, triglycerides,

LDL, and HDL obtained from laboratory tests
were analyzed using statistical software and
presented as mean ± SD. The distribution of the
obtained numerical data were also examined using
Shapiro Wilk test followed by Levene test in order
to determine the homogeneity of the data. If the
data were distibuted normally, then analysis of
variance (ANOVA) test was used, and Kruskall
Wallis test was used if the data were not normally
distributed. Differences were considered
significant when the probability value was less
than <0.05 (p<0.05).

Ethical clearance
This study was approved by the Ethical

Commision of the Islamic University of
Indonesia.

RESULTS

Before treatment, blood sampling was
performed for all groups in order to measure the
initial levels of total cholesterol, triglycerides, and
LDL. The mean levels of total cholesterol,
triglycerides, and LDL are presented in Table 1.
The data analysis began with the Shapiro Wilk
normality test and homogenity test using Levene
test. The Shapiro Wilk test for cholesterol,
triglycerides, and LDL indicated that the data had
a normal distribution. Therefore, one way Anova
test for cholesterol, triglyceride, and LDL was
applied. There were no signifficant differences
between groups before treatment (p>0.05). The
statistical analysis indicated that the subjects were
in similar condition before treatment.

During the first seven days, K1 rats were
given standard feed and water ad libitum, while

181

K2, P1, and P2 rats were treated with high-fat
feed and water ad libitum. On day 7, blood
sampling was again performed. Mean total
cholesterol, triglyceride and LDL levels on day
7 are shown in Table 2.

One way Anova test results for cholesterol
on day 7 showed significant differences between
groups (p=0.000). LSD post hoc test showed
significant differences (p <0.05) between groups
K1 in comparison with groups K2, P1 and P2.
The results of one way Anova test for triglyceride
levels on day 7 showed significant differences
between groups (p=0.000). LSD post hoc test
showed significant differences (p<0.05) between
group K1 in comparison with groups K2, P1 and
P2. Levels of LDL on day 7 also showed
significant differences between groups (p=0.008)
using Kruskal Wallis test (Table 2). Thus, the
administration of high-fat feeds in groups K2,
P1 and P2 led to significantly increased levels
of cholesterol, triglycerides, and LDL compared
with the negative control group (group K1).

The results for day 14 indicated that the
levels of cholesterol, and triglycerides, and LDL
showed significant differences between groups
(p=0.000; p=0.000; and p=0.001) (Table 3).

In this study, total cholesterol, triglycerides,
and LDL levels were highest in group K2 (the

group receiving high-fat feeds without honey).
This showed that the induction with high-fat
feeding in rats led to an increase in the levels of
total cholesterol, triglycerides, and LDL.

DISCUSSION

The induction with high-fat feeds causes an
increase in cholesterol synthesis and eventual
a c c u m u l a t i o n o f b l o o d c h o l e s t e r o l
(hypercholesterolemia). Increase in cholesterol
synthesis is facilitated by the enzyme HMGCoA-
reductase. Tomkin and Owens (12) state that the
decrease in cholesterol synthesis resulting from
inhibition of HMGCoA-reductase enzyme
activity reduces LDL levels and vice versa.
HMGCoA-reductase inhibitory activity is shown
b y s t a t i n s u s e d i n t h e t r e a t m e n t o f
hypercholesterolemia. If the cholesterol level in
the blood increases, LDL as a carrier of
cholesterol to cells throughout the body will also
increase. Furthermore, the accumulation of
cholesterol followed by free radical activity
causes oxidative damage to various tissues. In
addition, LDL is a substance that is readily
oxidized. Oxidized LDL binds to macrophages
and subsequently forms foam cells to cause
atherosclerotic lesions. The hypercholesterolemic

Table 1. Mean total cholesterol, triglyceride, and LDL levels by treatment groups at base line

K1: standard feed control group, K2: high fat feed control group, P1: high fat feeds for 7 days followed by high-fat feed
plus honey for the next 7 days, P2: high-fat feeds for 7 days followed by regular feed plus honey for the next 7 days. a Anova
test

Table 2. Mean total cholesterol, triglyceride, and LDL levels
by treatment groups after 7 days of treatment

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Nurmasitoh, Pramaningtyas Honey improves hypercholesterolemic

Table 3. Mean total cholesterol, triglyceride, and LDL levels by treatment groups
after 14 days of treatment

condition can lead to atherosclerosis, resulting in
complications such as myocardial infarction and
stroke.(12,13)

Uncontrolled hypercholesterolemia can
d e v e l o p i n t o c a r d i o v a s c u l a r d i s e a s e .
Hypercholesterolemia is strongly associated with
high total cholesterol and LDL levels in the
blood. High levels of total cholesterol and LDL
in the blood attract attention because they are
correlated with the incidence of coronary heart
disease.(1,3) Cardiovascular disease, such as
coronary heart disease, is one of the leading
causes of worldwide death. Mortality and
morbidity due to cardiovascular disease are
aggravated by a variety of risk factors, such as:
minimal physical activity and exercise, smoking,
poor dietary habits (diets high in fat and low in
fiber), alcoholism, and others.(14,15) Several
studies were developed to address the conditions
of hypercholesterolemia and maintain cholesterol
homeostasis. Cholesterol homeostasis is
maintained via a complex arrangement in terms
of absorption, anabolism, catabolism, and
excretion.(3,7,14)

According to previous studies, cholesterol
levels in the blood may be reduced through several
mechanisms. First, through competition with
intestinal Niemann Pick C1-like 1 (NPC1L1).
Around 1200-1700 mg of cholesterol enters the
lumen of the small intestine every day.
Approximately 300-500 mg of the cholesterol
comes from the diet and the rest comes from bile
acids. Cholesterol absorption in the small intestine
is mediated by NPC1L1. Once absorbed,
cholesterol is transported by NPC1L1 from the
lumen of the small intestine into the enterocytes.

Cholesterol is then converted into cholesteryl ester
(CE) by intestinal acyl-CoA cholesterol
acyltransferase 2 (ACAT2). In addition, the CE
will be packaged into chylomicrons by microsomal
triacylglycerol transport protein (MTP).
Eventually, chylomicrons head into the circulatory
system in the form of very-low density lipoprotein
(VLDL). Next, VLDL is degraded forming LDL
to deliver cholesterol to cells throughout the body.
The addition of substances that are structurally
similar to the NPC1L1 may reduce the levels of
cholesterol and LDL.(7,12,16)

The second mechanism is inhibition of
intestinal acyl-CoA cholesterol acyltransferase 2
(ACAT2). In mammals, ACAT2 is important for
lipid esterification in enterocytes and the liver to
produce cholesteryl ester which is useful for the
formation of chylomicrons and VLDL. Inhibition
of ACAT2 may lower total cholesterol and LDL.
The third mechanism is inhibition of 3-hydroxy-
3-methylglutaryl (HMG-CoA) reductase, which
is required in the biosynthesis of cholesterol. Thus,
inhibition of HMG-CoA reductase may inhibit the
cholesterol synthesis pathway so that the levels
of cholesterol and LDL in the blood are
reduced.(7,12,16)

The fourth mechanism is activation of LDL
receptors. Increasing the number and activity (up-
regulation) of LDL receptors may improve the
cholesterol-LDL clearance from the circulation.
Therefore, activation of the receptors may reduce
the levels of blood cholesterol and LDL. The fifth
mechanism is inhibition of bile acid reabsorption.
This is done through the binding of bile acids in
the intestine by certain substances to form
insoluble complex compounds that can not be

183

reabsorbed into the liver and are eventually
excreted. Inhibition of bile acid reabsorption
lowers cholesterol levels and increases the hepatic
synthesis of bile acids from cholesterol. The
increase in hepatic bile acid synthesis triggers
blood cholesterol influx into the liver. This is also
caused by LDL receptor activation due to
decreased hepatic cholesterol levels. Thus,
cholesterol levels and LDL in the blood will
decrease.(7,12,16)

The sixth mechanism is activation of
cytochrome P450 7A1 (CYP7A1) or cholesterol-
7á -hydroxylase. Activation (up-regulation) of
CYP7A1 is theoretically able to lower the
cholesterol in the liver, causing blood cholesterol
influx, and ultimately lowering cholesterol levels
in the blood.(7,12,16)

The last mechanism is inhibition of plasma
cholesteryl ester transporting protein (CETP).
CETP inhibition could be expected to lower blood
cholesterol and LDL levels because the
transportation of cholesteryl ester as a raw
material for making chylomicrons is
inhibited.(7,12,16)

Research on the mechanisms of cholesterol-
lowering therapy is important to determine the
substances and the working points which are
expected to lower total cholesterol and LDL levels
in the blood. One substance that is widely studied
as cholesterol-lowering agent is honey. Studies on
honey as an anti-hypercholesterolemia agent has
been widely developed, in both humans and
experimental animals with varying results. In the
present study, administration of honey in group
P1 and P2 resulted in significantly lower
cholesterol, triglycerides, and LDL levels in
comparison with those in group K2. Group P2
(the group receiving high-fat feeds followed by
normal feed plus honey) showed lower results
compared with group P1 (the group receiving
high-fat feeds followed by high-fat feeds plus
honey). In fact, total cholesterol, triglycerides, and
LDL levels of rats in group P2 were almost
identical with those in group K1 (the negative
control group receiving regular feed up to the end
of treatment).

Based on the data analysis perfomed in this
study, honey was proven to be able to reduce the
levels of cholesterol, triglycerides, and LDL. This
is in line with previous studies conducted on
various experimental animals. Nemoseck et al.(17)

stated that honey has many advantages that would
be promising in the world of health. One of the
benefits of honey that is described in their study
was to lose weight and lower blood triglyceride
levels in SD strain rats. In contrast, the other lipid
profile parameters were increased or were not
significantly different from those of the positive
control group or the group given additional
sucrose diet.

In addition, Alagwu et al.(13) suggest that the
administration of pure Nigerian honey can reduce
levels of blood LDL and total cholesterol in rats.
However, Alagwu’s research showed elevated
levels of triglycerides and VLDL in the blood.

Adnan et al.(1) also suggest that honey from
the acacia tree (desi kikar) may lower the levels
of triglycerides, total cholesterol, and LDL in diet-
induced hypercholesterolemic albino rats, as well
as in the group treated with simvastatin. The group
treated with a combination of acacia honey and
simvastatin showed greater reductions in lipid
profile, approaching the values in the negative
control group.

Research on the effects of different variants
of honey on the lipid profile has also been
conducted in humans. A research study by
Mushtaq et al.(18) mentions that the consumption
of 40 grams of honey was shown to improve lipid
profiles in obese adult subjects. Yaghoobi et al.(19)

also found that 70 grams of pure honey daily for
30 days was proven to reduce cardiovascular
disease risk factors and did not cause weight gain
in overweight or obese subjects. In addition,
studies on healthy young subjects with similar
dose and duration (70 grams daily for 4 weeks)
also showed a decrease in total cholesterol,
triglycerides, and LDL compared with the control
group.(20)

Honey is a nutrient-rich food that is
produced mainly by the honeybee (Apis mellifera
L.). Honey has been used by humans since

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Nurmasitoh, Pramaningtyas Honey improves hypercholesterolemic

ancient times. Various literature reports mention
that honey has antibacterial, antifungal, antiviral,
anti-inflammatory, antihypertensive, antioxidant,
antitumor, cardioprotective, hepatoprotective, and
hypoglycemic effects. Honey contains much
carbohydrate (glucose and fructose—85% content
of honey, sucrose, maltose, isomaltose, maltulose,
and others), vitamins (thiamine, riboflavin,
pyridoxine, vitamin A, niacin, pantothenic acid,
vitamin E, vitamin C), trace elements and minerals
(aluminum, arsenic, lithium, sulfur, iodine, cobalt,
sodium, calcium, potassium, magnesium,
phosphorus, zinc, copper, iron, selenium,
manganese, and others), polyphenols, organic
acids, amino acids (proline, glutamic acid, alanine,
phenylalanine, tyrosine, leucine, isoleucine, and
others), enzymes (glucose oxidase, invertase,
amylase, catalase, and acid phosphatase),
polyphenols (phenolic acids, flavonoids, phenolic
acid derivatives [quercetin, chrysin, galangin,
luteolin, kaempferol, apigenin, and others]), nitric
oxide (NO), and other substances. The
composition of the substances contained in honey
is influenced by the type of crop, climate, and
environmental conditions.(10,15, 21)

The hypotriglyceridemic effects of honey
may be caused by fermented nondigestible-
o l i g o s a c c h a r i d e s ( N D O s ) , s u c h a s
fructooligosaccharides (FOS) or other similar
carbohydrate isomaltulose which are contained
in honey. Nondigestible-oligosaccarides (NDOs)
are not hydrolyzed in the small intestine, but are
degraded by microflora in the colon. In the colon,
the degradation process will produce short chain
fatty acids that will affect the normal flora in
the colon. The normal flora of the colon is
important and has a beneficial effect on lipid
metabolism.(17) Polysaccharides that can not be
digested and short chain fatty acids resulting
from the fermentation process by the normal flora
will be able to reduce the levels of LDL
cholesterol in the blood by inhibiting HMG-CoA
reductase (7) and bile acid reabsorption into the
liver.(3,7,9,22) In addition, the normal flora species
which are beneficial and present in sufficient
numbers in the colon, such as Lactobacillus,

B i f i d o b a c t e r i u m , E n t e ro c o c c u s , a n d
Streptococcus, will be able to help lower blood
cholesterol levels by inhibiting the reabsorption
of bile acids into the liver.(7,22) Bogdanov et al.(9)

mention that honey can increase Lactobacillus
in the small intestine and colon.

The content of antioxidants in honey is also
predicted to have a beneficial effect on lipid
metabolism. Nevertheless, the content of
antioxidants in honey varies greatly depending on
the type of plant and its environment. Honey with
darker colors have higher antioxidant content than
the honey with a lighter color.(1,17) According to
Chen et al.,(7) antioxidant compounds, such as
quercetin, kaempferol, galangin, and various other
flavonoid compounds, are able to help lowering
LDL and cholesterol levels of blood. The
antioxidant compounds work by inhibiting
ACAT2, activating (up-regulating) LDL
receptors, and activating CYP7A1.

Chepulis and Starkey ( 2 3 ) found that
although there was an increase in HDL-
cholesterol in honey-fed rats compared with rats
fed sucrose or a sugar free diet, but there were
no other differences in lipid profiles. Nemoseck
et al.,(13) Alagwu et al.(17) and Majid et al.(20) also
suggest that the supplementation of honey will
increase the excretion of cholesterol through bile
acids thereby lowering blood cholesterol levels.
In addition to antioxidants, honey also contains
niacin. The content of niacin in honey could be
expected to lower blood cholesterol by inhibiting
the mobilization of deposits of triglycerides from
a d i p o s e c e l l s a n d t h r o u g h i n h i b i t i o n o f
diacylglycerol acyltransferase enzymes in liver
cells that are involved in cholesterol synthesis in
the HMG-CoA reductase pathway. (13,18,20)

Yaghoobi et al.(19) suggest that antioxidants in
honey, in addition to their role in lowering blood
c h o l e s t e r o l a n d L D L l e v e l s , a r e a l s o
advantageous by inhibiting the formation of
atherogenic plaques. Inhibition of atherogenic
plaque formation is also determined by a variety
of trace elements, minerals, vitamins, and NO
contained in honey. Some trace elements that
prevent the formation of atherogenic plaques are

185

copper and zinc. Meanwhile, some of the
vitamins contained in honey playing a role in
preventing oxidative stress and atherosclerotic
plaque formation are vitamin E (niacin), vitamin
C, vitamin B, and vitamin A.(20,24)

Honey is a food supplement with a complete
array of substances, of which some are potentially
of use as antihypercholesterolemic agents. Honey
contains several amino acid components.(15)

According to Chen et al.,(7) protein has the ability
to lower of blood cholesterol and LDL cholesterol
levels by inhibiting the reabsorption of bile acids,
inhibiting HMG-CoA reductase and activating
LDL receptors. In addition, honey also contains
calcium. Supplementation of calcium in the diet
is beneficial to lower cholesterol levels by
inhibiting intestinal NPC1L1, activating
CYP7A1, and increasing bile acid excretion via
feces.(7,25) Meanwhile, there are still many
substances in honey which have not been
scientifically proven to be beneficial in lowering
blood cholesterol levels.

In our study, the group treated with honey
followed by a normal diet had the lowest levels
of cholesterol, triglycerides, and LDL among all
treatment groups. Moreover, the level was almost
identical with that in the negative control group.
This indicates that in addition to adequate
therapy with antihypercholesterolemia agents,
dietary modification also has a positive effect
on the prevention of cardiovascular disease.(7,14,22)

This study had a limitation in the dose of
honey that should be given to rats. Further studies
should use a number of honey doses. The results
of this study may be useful to researchers and
clinicians as a reference in the use of honey
supplementation for those who are dyslipidemic.

CONCLUSIONS

Honey supplementation can reduce levels of
total cholesterol, triglycerides, and LDL in the
blood. Supplementation of honey accompanied by
a non-cholesterol diet can more effectively lower
total cholesterol, triglycerides, and LDL in the
blood.

CONFLICT OF INTEREST

There was no conflict of interest with other
institutions.

ACKNOWLEDGEMENT

This study was funded by the Faculty of
Medicine, Islamic University of Indonesia.

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