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Research Article
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Effect of Bitters on the Body Weight, Lipid Profile, Catalase and Lipid Peroxidation in Experimental Animals
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M.A. Alabi,
R.M. Sunday,
T. Olowokere,
F.A. Kareem
and
F. Osanaiye
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ABSTRACT
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This study was designed to investigate the potential effect of bitters on body
weight, lipid profile (total cholesterol, triglyceride, high density lipoprotein
(HDL)-cholesterol and low density lipoprotein (LDL)-cholesterol), catalase and
lipid peroxidation in the plasma and heart tissue of Albino rats. Three brands
of bitters (Yoyo, Swedish and Living bitters) were administered daily for thirty-two
days through oral rout to age matched twenty eight Albino rats of both sexes.
The rats were grouped into four with seven rats per group. The control group
was administered normal saline, the treatment groups were administered with
Yoyo, Swedish and Living bitters respectively (15 mL kg-1). The animals
were sacrificed after thirty-two days. There was a significant (p<0.05) decrease
in body weight, a reduction in cardiac total cholesterol, lipid peroxidation,
triglyceride and LDL-cholesterol levels and an increase in catalase activity.
There was also a significant reduction in plasma total cholesterol and triglyceride
levels but significant increase in plasma catalase activity. Hence, bitters
could be recommended to be taken as digestive, as it may help to reduced the
body weight, cardiac total cholesterol, triglyceride, LDL-cholesterol levels,
lipid peroxidation level and increase catalase activity.
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Received: January 26, 2013;
Accepted: March 13, 2013;
Published: May 06, 2013
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INTRODUCTION
Bitters are prescription and natural remedies that are commonly used in developing
countries as a cure for indigestion and other stomach ailments and for treatment
of various diseases. Its use has been on the increase recently without investigations/examinations
of the possible toxic effect or metabolic alteration. Studies or reports over
the last two and a half decades have constantly demonstrate increasing use of
herbal remedies in both developing countries and in developed countries where
modern medicines are predominantly used medicine as the most common form of
alternative medicine and is used by about 60-80% of the world (Bussmann
et al., 2010). Bitters include but are not limited to: Gentian root
(Gentiana spp.) (Jaeschke et al., 2010),
Aloe (Aloe vera syn. A. arbadensis) (Reynolds
and Dweck, 1999), Wormwood (Artemisia absinthium) (Kharoubi
et al., 2008) from which absinthe was made, Dandelion root (Taraxacum
officinale) (Choi et al., 2010), Angelica
root (Angelica archangelica) (Sarker and Nahar, 2004),
Senna leaves (Cassia senna) (El-Kamali and El-Amir,
2010), Zedoary root (Curcuma zedoaria) (Krishnamoorthy
et al., 2009), Myrhh (Commiphora molmol) (Tariq
et al., 1985), Cinchona bark (Cinchona spp.) (Schulz
and Albroscheit, 1988), Turmeric (Curcuma longa syn. C. domestica)
(Kim and Kim, 2010), Shitetta (Swertia chirata
syn. Ophelia chirata) (Verma et al., 2008)
and Saffron (Crocus sativa) (Motamedi et al.,
2010). Bitters generally have been reported to prevent kidney and bladder
infections, help to regulate blood pressure and dilate arteries, facilitate
digestion, prevent disorder like ulcers, gastritis, insomnia, stress and depression
and prevent overweight and excess body fat (Ogbonnia et
al., 2010). The medicinal use of the extract from these plants are well
documented but there exist little or no information as regard the effect on
lipid profile (total cholesterol, triglyceride, HDL-cholesterol and LDL-cholesterol),
catalase and lipid peroxidation (thiobarbituric acid reactive substances (TBARS))
of the heart by the extract these plants when used in combination (Krishnamoorthy
et al., 2009). The herbs are used in combination creates the desired
synergistic effect (Krishnamoorthy et al., 2009).
The aim of this study was to investigate the potential effect of bitters on
the lipid profile (total cholesterol, triglyceride, HDL-cholesterol and LDL-cholesterol),
catalase and lipid peroxidation (TBARS) in the plasma and heart tissue of Albino
rats.
MATERIALS AND METHODS
Food supplements: Yoyo bitters were obtained from Ablatt Pharmaceuticals
Limited, Nigeria, Swedish Bitters was purchased from Swedish Bitters Herb Company,
USA and Living bitters from African Angel, USA.
Reagents and chemicals: Bovine serum albumin, sodium hydroxide, copper
sulphate, sodium potassium tartarate, potassium iodide, sodium dihydrogen phosphate,
disodium hydrogen phosphate and sodium chloride were products of Aldric Chemicals,
UK. All other chemicals are of analytical grade.
Animal: Age matched 28 Albino rats of both sexes which were purchased
from the Physiology Department of the University of Ibadan, Ibadan, Oyo State,
Nigeria. The rats were randomly grouped into four (YB-animal fed with Yoyo Bitters,
SB-Swedish Bitters, LB-Living Bitters and CT as control) and kept in wooden
cages, which were well ventilated. The animal were allowed to acclimatize to
their new environment for two weeks and fed with normal rat chow (Guinea Feed
Ltd, Nigeria) and water ad libitum. After two weeks of acclimatization,
the four groups of animals were administered, Yoyo Bitters (YB), Swedish Bitters
(SB), Living Bitters (LB) (15 mL kg-1 b.wt.) and normal saline (CT)
through oral rout once daily, in addition to the normal rat chow and water for
32 days (about 5 weeks).
Body weight: At least twice every week the body weight of each animal
was measured.
Sample collection: After 32 days, the rats were sacrificed by cervical
decapitation under the influence of diethyl ether anaesthesia. Blood samples
were collected from each animal by cardiac puncture with sterile needle and
transferred to heparinised tubes, centrifuged at 5,000 rpm for 10 min and the
supernatant and plasma, was separated and stored at a temperature of -4°C
until it is required for assaying. The heart was dissected out, washed in ice-cold
saline, dried and weighed. The heart was then homogenized in normal saline solution
in the ratio 1:4 (1 mg of heart tissue to 4 mL of normal saline solution) and
the homogenate was centrifuge at 4000 rpm for 5 min while the supernatant was
separated and stored at a temperature of -4°C until it is required for assaying.
Biochemical analysis: Triglyceride level was determined by the method
of Mochin and Leyva (1984), total cholesterol and HDL-cholesterol
levels were determined by the method of De Hoff et al.
(1978) and LDL-cholesterol level was calculated by the method of Nauck
et al. (2000). Catalase activity was determined by the method of
Johansson and Borg (1988) and TBARS were determined by
the method of Buege and Aust (1978).
Statistical analysis: All the results were expressed as Mean±Standard
Error of Mean (SEM) for animal in each group. All the grouped data were statistically
evaluated using SPSS 15.0 software. Hypothesis testing methods included One-way
Analysis of Variance (ANOVA) and subsequent comparisons among groups were made
using Duncans Multiple Range
Test (DMRT). Statistical significance was set at p<0.05.
RESULTS
Effect of bitters on body weight: There was a significant decrease in
body weight of the animal fed with Yoyo bitters whereas a significant reduction
was observed in animal fed with Swedish and Living bitters from 15 days as shown
in Fig. 1.
Effect of bitters on lipid profile: The lipid profile result (Table
1, 2) shows a significant reduction in the total cholesterol
level of the plasma and heart tissue, respectively in all the animal fed with
Yoyo bitters (162.78±2.19, 653.97±56.07), Swedish bitters (161.52±2.52,
703.50±35.98) and Living bitter (164.05±2.52, 668.54±29.80)
when compared with the control (181.71±5.55, 1123.78±212.66) at
p<0.05; a significant reduction in the triglyceride level of the heart tissue
in animal fed with Yoyo bitters (1519.77±111.59), Swedish bitters (1999.34±43.51)
and Living bitters (1964.09±116.67) when compared with the control (3219.67±583.12)
at p<0.05 whereas no significant difference was observed in the plasma triglyceride
level in the animal fed with Yoyo bitters (89.64±9.78), Swedish bitters
(88.55±5.18) and Living bitter (89.66±3.79) when compared with
the control (88.17±0.76) at p<0.05; a significant elevation in LDL-cholesterol
level of the plasma and heart tissue, respectively in the animal fed with Yoyo
bitters (39.32±2.22, 181.44±24.91), Swedish bitters (39.18±2.29,
135.72±25.57) and Living bitter (33.71±6.19, 114.10±25.35)
when compared with the control (64.76±10.77, 282.99±54.82) at
p<0.05; but no significant difference in HDL-cholesterol level of the plasma
and heart tissue respectively in the animal fed with Yoyo bitters (105.53±2.44,
168.37±34.91), Swedish bitters (104.63±1.77, 167.91±35.09)
and Living bitter (112.40±5.98, 162.26±37.46) when compared with
the control (99.31±10.11, 196.86±23.74) at p<0.05.
Effect of bitters on catalase and lipid peroxidation: The catalase activity
and lipid peroxidation results (Table 3, 4)
also showed a significance increase in catalase activity of the plasma and heart
tissue respectively in animal fed with Yoyo bitters (8.50±0.03, 8.48±0.11),
Swedish bitters (8.34±0.06, 8.63±0.06) and Living bitter (8.60±0.05,
8.51±0.10) when compared with the control (8.07±0.03, 8.07±0.13)
at p<0.05 but there was a decrease in TBARS level of the plasma and heart
tissue, respectively in the animal fed with Yoyo bitters (12019.23±3582.25,
4.33±0.80), Swedish bitters (15000.00±1453.17, 4.74±0.83)
and Living bitter (8226.49±1830.29, 4.17±0.64) when compared with
the control (25213.68±2230.83, 3.63±0.94) at p<0.05.
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Fig. 1: |
Effect of Yoyo, Swedish and Living bitters on body weight
of experimental animal when compared with control |
Table 1: |
Effect of bitters on plasma lipid profile |
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All value are expressed as Mean±SEM. Values with different
superscript indicate significant difference at p<0.05 |
Table 2: |
Effect of bitters on cardiac lipid profile |
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All value are expressed as Mean±SEM. Values with different
superscript indicate significant difference at p<0.05 |
Table 3: |
Effect of bitters on plasma catalase activity and lipid peroxidation |
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All value are expressed as Mean±SEM. Values with different
superscript indicate significant difference at p<0.05 |
Table 4: |
Effect of bitters on cardiac catalase activity and lipid
peroxidation |
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All value are expressed as Mean±SEM. Values with different
superscript indicate significant difference at p<0.05 |
DISCUSSION
The present study was designed to investigate the effect of bitters on the
body weight, lipid profile (total cholesterol, triglyceride, HDL-cholesterol
and LDL-cholesterol), catalase and lipid peroxidation (TBARS) in the plasma
and heart tissue of Albino rats.
The study showed that all experimental animals exerted a significant decrease
in body weight in the experimental animal fed with bitters (Yoyo, Swedish and
Living bitters) when compared with control. Intake of bitters was also shown
to lower total cholesterol and LDL-cholesterol levels in the plasma and heart
tissue; and triglyceride in the heart tissue. Lowering total cholesterol, triglyceride
and LDL-cholesterol levels of the plasma and heart tissue reduces the risk of
hypercholesterolemia and hyperlipidemia that may precipitate into coronary atherosclerosis
and other related cardiovascular diseases (Witztum and
Steinberg, 2001). This could be as a result of the fact that bitter helps
in oxidation of total cholesterol and triglyceride plus reduction in LDL-cholesterol
level. The study also showed an increase in catalase activity and a decrease
in the level of lipid peroxidation products (TBARS) in both the plasma and the
heart tissue. Oxidative stress is one of the causative factors that link hypercholesterolemia
with atherogenesis. Malondialdehyde (MDA), the product of lipid peroxidation,
is an index of the level of oxygen free radicals. The increased catalase activity
help to scavenge oxygen free radicals (Jaeschke et al.,
2010).
CONCLUSION
In conclusion, the present results show that moderate consumption of bitters
may lower body weight, total cholesterol, LDL-cholesterol and lipid peroxidation
and increase catalase activity.
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