Antiobesity effect of Phytolacca berry in rats

Abstract

Extract of berries from Phytolacca americana L. plant (Phytolacca berry) is widely used in alternative systems of medicine i.e. in homeopathy for the treatment of obesity. A study was made to evaluate its anti-obesity effect in the rat model. The experiement was conducted in rats using the diet-induced obesity model in which cafeteria and atherogenic diets were used. The activity was tested by measuring body weight, glucose, total cholesterol and triglyceride levels in comparison with a control group. The results revealed that the Phytolacca berry extract had significant anti-obesity activity by reducing excess body weight, and cholesterol and triglyceride concentrations. This activity may be due to its appetite suppressant activity and also by increasing the metabolic rate. Further studies are needed to be conducted to prove its utility in humans.

Full text

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Antiobesity effect of Phytolacca berry in rats
G. Ravi Kiran1, Akondi Butchi Raju2*
1Department of Pharmacology, St. Peters Institute of Pharmaceutical Sciences, Hanamkonda, Warangal, India
2Department of Pharmacology, Ibn Sina National College for Medical Studies, Jeddah, Saudi Arabia
*Corresponding author, E-mail: drraju2020@gmail.com
Abstract
Extract of berries from Phytolacca americana L. plant (Phytolacca berry) is widely used in alternative systems of medicine i.e. in
homeopathy for the treatment of obesity. A study was made to evaluate its anti-obesity effect in the rat model. e experiement was
conducted in rats using the diet-induced obesity model in which cafeteria and atherogenic diets were used. e activity was tested
by measuring body weight, glucose, total cholesterol and triglyceride levels in comparison with a control group. e results revealed
that the Phytolacca berry extract had significant anti-obesity activity by reducing excess body weight, and cholesterol and triglyceride
concentrations. is activity may be due to its appetite suppressant activity and also by increasing the metabolic rate. Further studies are
needed to be conducted to prove its utility in humans.
Key words: atherosclerosis, homoeopathy, obesity, Phytolacca americana, Phytolacca berry, rat model.
Abbreviations: AD, atherogenic diet; CD, cafeteria diet.
Environmental and Experimental Biology (2014) 12: 95–99 Original Paper
Introduction
Maintaining energy homeostasis is fundamental for
survival. However, obesity is due to over-nutrition and an
increasing worldwide public health problem (Friedman
2000; Wilborn et al. 2005). e World Health Organization
recognized the obesity epidemic as one of the top 10 global
health problems. In developed countries it is estimated that
5% of total health costs are related to obesity (Kopelman
2000; Odgen 2002) and is oen considered a problem of
the belly rather than of the brain. Epidemiological studies
from India suggest a rise in morbid obesity, close to 5%
(Misra 2001). Obesity is the excess accumulation of fat in
the body and an imbalance in energy intake and energy
expenditure, which is the most common nutritional
disorder in the developed world and is considered to be a
risk factor associated with the development of the major
human diseases. Various factors may lead to obesity, such as
sedentary life style, increased intake of high calorie (energy
and fat) food, genetic determinants and psychologic and
behavioural determinants (Wilborn et al. 2005).
Phytolacca americana L. (Phytolacca berry) is a common
perennial native plant found in Northern and Central
North America. It is widely used to treat obesity due to
its appetite suppressant activity, and hypocholesterolemic
and excess body weight reducing properties (Haider
et al. 2006). Phytochemical investigations of the plant
revealed the presence of triterpene saponins, triterpene
alcohols, betacyanins and lignanes etc. (Wang 2008). No
extensive pharmacological studies have been conducted
on Phytolacca berry. erefore, the aim the present study
was to conduct an evaluation of the antiobesity activity of
Phytolacca berry extract on diet-induced obese rats.
Materials and methods
Forty two female wistar rats (180 to 220 g) were obtained
from the animal house facility of St. Peters Institute of
Pharmaceutical Sciences, Warangal, India. ey were
divided into seven groups and housed in polypropylene
cages under standard laboratory conditions at a room
temperature 23 ± 2 °C with 12/12 h light/dark cycle. e
animals were provided with pellet chow and water ad
libitum. e study protocol was approved by Institutional
Animal Ethical Committee. Standard kits used for the
estimation of total cholesterol, triglyceride, glucose and
total protein were purchased from Crest Biosystems, Goa,
India. Cholesterol and cholic acid were from SD Fine
Chemicals (Mumbai, India), and Lard oil from Sigma Chem.
Co. (USA). e entire experimental work was conducted
in the department of pharmacology, St. Peters Institute of
Pharmaceutical Sciences, Warangal, Andhra Pradesh, India.
Phytolacca berry tablets were obtained from Dr.
Willmar Schwabe India Pvt. Ltd. (Noida, India). Each tablet
contained 20% of the Phytolacca berry extract.
Obesity was induced by using two types of diets:
cafeteria diet (CD; Harris 1993) and atherogenic diet (AD;
Jiao et al. 1991). Cafeteria diet was a high carbohydrate/
sugar diet and consisted of condensed milk (40 g), bread
(40 g), chocolate (15 g) biscuits (30 g), dried coconut (30 g),
cheese (40 g) and boiled potatoes (50 g). e atherogenic
diet consisted of 1% cholesterol, 0.5% cholic acid, and 5%
E  E B ISSN -

G.R. Kiran, A.B. Raju
96
lard oil. ese high calorie diets were provided to each rat
through oral ingestion along with a normal pellet chow.
e animals were divided in seven groups containing
six animals each. Group I served as control and received
only normal pellet diet; Group II received AD; Group III
received AD and Phytolacca Berry extract high dose (2.850
mg kg–1); Group IV received AD and Phytolaccca Berry
low dose (0.285 mg kg–1); Group V received CD; Group VI
received CD and Phytolacca Berry high dose; and Group
VII received CD and Phytolacca berry low dose for 40 days.
e normal pellet diet was provided to all groups of rats ad
libitum. Cafeteria and atherogenic diets were orally fed to
rats twice a day in the quantity of 10 mL kg–1.
Evaluation of physiological parameters were performed
according to Kaur and Kulkarni (2000).
e rats of different groups were weighed initially on
day 1 and then on alternate days for 40 days and recorded.
e body temperature of the rats in different groups was
recorded on day 39 using a digital telethermometer (Inco
Instruments and Chemicals, Ambala, India) for a period of
3 h at 0, 30, 60, 90, 120 and 180 min using contact time of
1 min.
Locomotor activity was recorded on day 40 for all
the treatment groups using the open field behavior test
apparatus aer 30 min of the administration of test drug.
e apparatus consisted of a circular wooden area of 75 cm
and a wall with a height of 25 cm. e test was performed
by placing the rat in the middle of the apparatus and
recorded the frequency of rearing and grooming activity
and ambulatory activity for 5 min.
Aer exsanguination of rats by cervical dislocation,
different organs (kidney, liver, heart, spleen) and fat pads
(mesenteric, ovarian, perirenal and uterine) were removed
and weighed.
Glucose was determined according to Trinder
(1969). e blood was collected from rats immediately
aer exsanguinations and serum was separated using
centrifugation. In all the groups on day 41 the serum
glucose was estimated by using GOD/POD method with
a biochemical kit manufactured by Crest Biosystems (Goa,
India).
e separated serum was subjected to biochemical
analysis and total cholesterol (CHOD/PAP method; Allain
1974), triglycerides (GPO/PAP method; Bucolo et al. 1973)
and total protein (Biuret method; Gornall et al. 1949) were
determined.
e results were expressed as means ± SE. Comparisions
between treatment and control groups were performed by
analysis of variance (ANOVA) followed by the Bonferroni
multiple comparison test. e statistical level of significance
was P < 0.05.
Results and discussion
Obesity results from dysregulation between energy
intake and expenditure. It is believed to be associated
with numerous diseases including hyperlipidemia,
hypercholesterolemia and type 2 diabetes (Friedman 2000).
ere are many drugs currently used for the treatment of
obesity, but due to the various complications which they
produce, research has been carried out on identifying
phytochemicals from traditional medicinal plants that
can provide a safer therapy for obesity. is will open new
research avenues for the future (Haider et al. 2006).
In this study, Phytolacca berry tablets were tested. ey
contain an extract of Phytolacca americana native to North
America and are widely used as a homoepathic medicine to
treat obesity. is study provides a suitable explanation for
the anti-obesity activity of Phytolacca berry. Animal models
of obesity have been reported to bear close resemblance to
human obesity (Sclafani et al. 1976).
In this study obesity was induced by two types of diets:
cafeteria diet (Group II), which was a high carbohydrate
diet, and atherogenic diet (Group III), which was a high fat
diet given along with a pellet chow.
ere was a significant increase in body weight in
the group of animals fed with cafeteria diet (CD) and
atherogenic diet (AD) compared with the control group.
Fig. 1. Effect of Phytolacca berry tablets on body weight of rats
fed with atherogenic diet (A) and cafeteria diet (B) for 40 days.
A.D., atherogenic diet, C.D., cafeteria diet; T.D. H.D., test drug
high dose, T.D. L.D., test drug low dose.
A
B

Antiobesity effect of Phytolacca berry
97
Treatment with the test drug caused significant decrease in
body weight when compared to the control group and also
significantly effective in reducing weight in the cafeteria
and atherogenic diet group animals (Fig. 1).
ere was a significant increase in ambulatory activity
and rearing activity in the cafeteria fed group compared
to control group. Treatment with the test drug resulted in
promotion of ambulatory activity and rearing activity and
there was an increase in ambulatory activity in AD with
Phytolacca berry treatment. ere was a significant increase
in grooming activity in AD and CD drug treatment groups
as compared to that for the control group (Table 1).
Treatment with the test drug significantly increased
body temperature in both cafeteria and atherogenic diet
groups (Table 2). is indicates that Phytolacca berry
increased the metabolic rate in the body, which ultimately
resulted in lowering of fat stores.
Administration of CD and AD increased the weight
of organs and increased fat accumulation in uterine and
perirenal fat pads. Treatment with the test drug in the
cafeteria diet group reduced fat accumulation in uterine
and perirenal fat pads (Table 3).
Treatment with the test drug showed increased glucose
level, compared to that in the control group especially in
CD animals im the high dose drug group, but it significantly
decreased total cholesterol and triglyceride levels in CD
and AD groups, which had high cholesterol and triglyceride
concentration. compared to those in control group animals.
Animals fed with the cafeteria diet had higher total protein
content compared to that in the control, and treatment with
Table 1. Effect of Phytolacca berry on open field behavior of rats fed on different diets. Values are means ± SE. *, significant difference to
control; a, significant difference to atherogenic diet group; b, significant difference to cafeteria diet group (P < 0.05)
No Treatment Frequency of open field behavior
Ambulation Grooming Rearing
1 Control 74.67 ± 3.33 8.33 ± 0.33 13.17 ± 1.42
2 Atherogenic diet 59.00 ± 0.78 5.66 ± 0.42 23.17 ± 1.99*
3 Atherogenic diet + test drug high dose 93.00 ± 2.30* 10.83 ± 0.79a 38.00 ± 2.69*
4 Atherogenic diet + test drug low dose 80.33 ± 4.86a 8.33 ± 1.08 19.5 ± 2.2
5 Cafeteria diet 84.17 ± 3.10 5.33 ± 1.25 41.17 ± 1.53*
6 Cafeteria diet + test drug high dose 96.33 ± 4.10* 12.17 ± 1.24b 27.33 ± 2.89*
7 Cafeteria diet + test drug low dose 64.5 ± 4.68b 11.83 ± 1.83b 30.67 ± 1.74*
Table 2. Effect of Phytolacca berry on body temperature (°C) of rats fed on different diets. Values are means ± SE. *, significant difference
(P < 0.05) to control group
No Treatment Body temperature at time (min)
0 30 60 90 120 180
1 Control 38.08 ± 0.04 38.73 ± 0.17 38.67 ± 0.19 38.97 ± 0.03 38.47 ± 0.16 38.43 ± 0.13
2 AD 39.92 ± 0.08* 39.75 ± 0.17* 39.5 ± 0.18* 39.87 ± 0.12 40.0 ± 0.0* 39.75 ± 0.17*
3 AD + test drug high dose 38.08 ± 0.04 39.83 ± 0.16* 40.10 ± 0.06* 40.23 ± 0.08* 39.58 ± 0.20* 38.5 ± 0.18
4 AD+ test drug low dose 39.17 ± 0.40* 39.75 ± 0.17* 39.8 ± 0.16* 39.67 ± 0.21 39.25 ± 0.17 39.20 ± 0.38
5 CD 38.15 ± 0.20 38.27 ± 0.16 38.23 ± 0.15 38.30 ± 0.15 37.92 ± 0.20 38.17 ± 0.16
6 CD + test drug high dose 38.77 ± 0.30 38.8 ± 0.19 39.23 ± 0.40 38.83 ± 0.30 38.67 ± 0.21 38.25 ± 0.25
7 CD + test drug low dose 37.83 ± 0.41 39.0 ± 0.36 39.08 ± 0.32* 38.9 ± 0.19 38.6 ± 0.20 38.25 ± 0.30
Table 4. Effect of Phytolacca berry on various biochemical parameters in rats fed on different diets. Values are mean ± SE. *, significant
difference to control; a, significant difference to AD; b, significant difference to CD (P < 0.05)
No Treatment Biochemical parameters (mg dL–1)
Glucose Total cholesterol Triglycerides Total protein
1 Control 45.33 ± 4.716 84.67 ± 5.19 93.67 ± 3.06 7.31 ± 0.28
2 AD 65.33 ± 8.22 100.3 ± 4.36 111.7 ± 7.46 8.25 ± 0.44
3 AD + test drug high dose 63.33 ± 12.49 66.5 ± 5.84a 78.17 ± 5.91 8.73 ± 0.28
4 AD + test drug low dose 47.67 ± 6.20 80.67 ± 3.82 93.83 ± 8.35 8.08 ± 0.24
5 CD 92.67 ± 3.31* 88.33 ± 6.46 165.8 ± 4.9* 10.65 ± 0.50*
6 CD + test drug high dose 110.3 ± 21.01* 53.83 ± 4.93*ab 123.8 ± 5.43*b 9.35 ± 0.41*
7 CD + test drug low dose 97.67 ± 5.61* 62.5 ± 2.30*ab 156.5 ± 9.55* 9.45 ± 0.33*

98
G.R. Kiran, A.B. Raju
the test drug lowered the total protein content in cafeteria
fed animals (Table 4).
e major building block for the synthesis of triglycerides,
in tissues other than adipose tissue, is glycerol. Adipocytes
lack glycerol kinase, therefore, dihydroxyacetone phosphate
produced during glycolysis is the precursor for triglyceride
synthesis in adipose tissue. is means that adipocytes
must have glucose to oxidize in order to store fatty acids in
the form of triglycerides. Dihydroxyacetone phosphate can
also serve as a backbone precursor for triglyceride synthesis
in tissues other than adipose. Upon treatment with the
test drug, triglyceride levels were significantly reduced.
Total protein concentration was increased in cafeteria diet
fed animals. Treatment with the test drug decreased the
concentration of total protein in the cafeteria group.
e results of the study showed that rats fed with a
variety of highly palatable, energy rich, high carbohydrate
cafeteria foods elicited significant increase in body weight
and fat pad mass. Cafeteria diet has previously been
reported to increase energy intake and cause obesity in
humans and animals (Rothwell et al. 1983; Bull 1988). In
this study, atherogenic diet-fed rats also exhibited increased
body weight along with a corresponding rise in cholesterol
levels. us, it was found that the test drug (Phytolacca
berry tablets) used with higher dose had significant effect
on obesity. e weight-reducing property of this test drug
may be due to its thermogenic property, by increasing
metabolic activity and appetite suppressant activity, which
controls feed intake.
In conclusion, alternative medicine has been proved as
a good source for identification of lead compounds. In the
current experimental study, antiobesity effect of phytolacca
berry was conducted on rats, and the results were promising.
Parameters like weight of the animal, biochemical markers
and fat pad weights showed the efficacy of Phytolacca
berry extract tablets against obesity in rats. Further studies
need to be conducted to evaluate its efficacy in the human
population.
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Table 3. Effect of Phytolacca berry on organ and fat pad weights in rats fed on cafeteria and atherogenic diets. Values are means ± SE. *, significant difference to control; b, significant
difference to cafeteria diet (P < 0.05)
No Treatment Organ weight (g) Fat pad weight (g)
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1 Control 0.91 ± 0.09 6.63 ± 0.66 1.12 ± 0.27 0.72 ± 0.05 0.76 ± 0.02 1.23 ± 0.32 2.7 ± 0.81 1.82 ± 0.79 0.64 ± 0.20
2 AD 0.9 ± 0.07 7.59 ± 0.26 1.15 ± 0.09 0.70 ± 0.05 0.67 ± 0.11 1.69 ± 0.20 4.23 ± 0.55* 1.91 ± 0.30 0.52 ± 0.07
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4 AD + test drug low dose 0.93 ± 0.04 7.28 ± 0.16 1.08 ± 0.10 0.72 ± 0.02 0.72 ± 0.02 2.07 ± 0.18 3.10 ± 0.51b 2.13 ± 0.23 0.52 ± 0.06
5 CD 1.14 ± 0.21 6.4 ± 0.35 1.28 ± 0.19 0.84 ± 0.04 0.76 ± 0.01 1.72 ± 0.42 5.45 ± 0.69* 1.54 ± 0.43 1.24 ± 0.21
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7 CD+ test drug low dose 1.03 ± 0.05 6.44 ± 0.34 1.23 ± 0.16 0.89 ± 0.04 1.24 ± 0.24 2.29 ± 0.05 5.77 ± 0.92* 1.71 ± 0.33 1.45 ± 0.34

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Received 28 May 2014; received in revised form 11 July 2014; accepted 2 September 2014