International Research Journal of Biochemistry and Biotechnology

IRJBB

Vol. 2(2), pp. 028-036, December, 2015. www.premierpublishers.org, ISSN: 2167-0438x

Research Article

Serum lipid profile and liver enzymes of Rats fed

Lageneria sphaerica (Wild bottle gourd) supplemented
diet
Chinyere G.C. 1*, Ikpoh J.C.2, Osuocha K.U3
1*,2,3

Department of Biochemistry, Abia State University Uturu, Abia State, Nigeria.

The present study evaluated serum lipid profiles and liver enzymes activities of male albino rats
fed Lageneria sphaerica supplemented diet. A total of forty two rats were used for this study.
The rats were separated into three groups of thirteen rats each. Group I was fed with normal rat
chow, group II was fed with 60% sample and 40% normal rat chow while group III was fed with
30% sample and 70% normal rat chow respectively. There was a significant decrease in weight
(P<0.05) with increased sample supplementation in relation to control. From the results
obtained, serum liver enzyme activities increased significantly (P<0.05) in group II animals as
feeding period progressed. Values obtained for these serum liver enzymes were Aspartate
aminotransferase (AST), 31.883.47U/L to 37.383.42U/L, Alanine aminotransferase (ALT)
12.25260U/L to 17.882.25U/L and Alkaline phosphatase (ALP) 519.4795.01U/L to
483.8662.09U/L respectively. Similar results were obtained for the group III rats. There were
also significantly (P<0.05) increased serum triglyceride and total protein while serum
cholesterol reduced significantly (P<0.05) in the test groups. From the results obtained,
Lageneria sphaerica seed is a probable toxic seed and may not be suitable for consumption
unless appropriate detoxifying methods are applied.

Keywords: Lageneria sphaerica, serum, lipids, liver enzymes

INTRODUCTION
Lageneria sphaerica is commonly known as wild melon
and are found in low lying areas in the tropical regions of
the world. The bitter taste of the fruits are attributed to
elastase activity (Ojiako and Igwe, 2007). This plant is a
perennial with a woody rootstock found naturally in
tropical and Southern Africa (Ellof et al., 2007).It grows in
full sun and semi-shade in forest margins, river-banks
and in dry river beds(Jeffrey, 1978). There are variable
reports on the medicinal uses of parts of this plant
(Fagbemi and Oshodi, 1991). The leaves of different
species of Lageneria sphaerica have been reportedly
used as food condiments while the Cucurbitacaes
oilseeds have been employed in domestic activities
(Odoemena, 2005).These oilseeds are good sources of
lipids and proteins with defatted cakes capable of being

used as a protein supplement in human nutrition. Many

workers have cited different medicinal uses of this plant
in various countries (Saha et al., 2011; Anaga, 2011;
Shah et al., 2010; Mohale et al., 2009; Gangwal et al.,
2008). Similarly the phytochemical screening of the fruits
revealed the absence of tannings (Chinyere et al., 2009)
but good amount of nutrients that compared favourably
with those of melon and other seeds of the same species.

*Corresponding Author: Dr. Chinyere G.C., Department

of Biochemistry, Abia State University, Uturu, Abia State,
Nigeria. Email: gcchinyere@yahoo.com

Serum lipid profile and liver enzymes of Rats fed Lageneria sphaerica (Wild bottle gourd) supplemented diet

Chinyere et al.

028

The liver contains thousands of enzymes of which are

also present in serum in very low concentrations (Sarada
and Madhanankutty, 1990). These enzymes have no
known functions in the serum other than provide
information about hepatic state and disorders. These
enzymes are distributed in plasma and intestinal fluid and
have characteristic half-lives usually measured in days.
The elevation of any of these enzymes activities in the
serum reflect increased rate of entrance into the serum
from damaged liver cells. Amongst these enzymes are
Aspartate
anminotransferase
(AST),
Alanine
aminotransferase (ALT), Alkaline phosphatase (ALP) and
others. These enzymes in conjunction with albumin and
conjugated bilirubin tests are used in determining the
functional integrity of the liver. The serum levels of these
enzymes are especially altered in hepatocellular diseases
that are acute in forms. They are therefore referred to as
hepatocellular enzymes (Ghadi, 2000). Elevated serum
transaminases levels are typically associated with acute
hepatocellular necrosis and reflect the release of
enzymes from the cytoplasm of dying cells. Other
features such as fatty degeneration, infiltration by
inflammatory cells are variable and reflect the severity of
injury (Kaplowitz, 1992).The most commonly used marker
enzymes of hepatic injury are AST and ALT. Hepatic
necrosis in acute hepatitis, toxic injury or ischemic injury
results in the leakage of these enzymes into circulation
(Healey et al., 1995)
Cholesterol belongs to a class of lipids known as derived
lipids (Ononogbu, 1998).The biosynthesis of cholesterol
predominantly takes place in the liver and intestinal
mucosa but almost all cells synthesize it. It is a
constituent of many membranes, essential in the
synthesis of bile acids and steroid hormones (Murray et
al., 2005). It circulates in blood as cholesterol ester
bound to beta lipoprotein. The measurement of the serum
cholesterol, triacylglycerol and lipoprotein levels are
important in examining the metabolism of lipids. Changes
in serum cholesterol level reflect disorders of liver
function. Serum cholesterol level is increased in
obstructive
jaundice,
diabetes
mellitus
and
hypothyroidism (Sarada and Madhanyankutty, 1990).
Histological examination of tissues samples are important
as other ways of diagnosing diseases (Kiceniuk et al.,
1982).Series of hepathological changes may be seen
when organisms are exposed to toxic agents. The degree
of toxicity can therefore be observed through the extent
of damage by the toxic substances to the tissues and
hence the need for liver and tissue biopsy in the
assessment and confirmation of the state of the liver and
other tissues (Brunt et al., 1984).
The seeds of Lageneria sphaerica are widely used by
many countries of the world for nutritional and medicinal
purposes. Scanty literature exists on the effect of intake
of this plants seed on biochemical parameters. In our
earlier communication, we evaluated the nutritional
qualities of Lageneria sphaerical seed (Chinyere et al.,

2009). The present study is aimed at evaluating the effect

of its intake as a food supplement on serum lipids and
liver enzymes profiles using albino rats models. The
results obtained here will shade light into a possible use
of this plants seed as a constituent of livestock feeds and
for human nutrition.
Method of analysis
Animals
Thirty Six male albino rats weighing 112-118g were
purchased from the Animal house, Department of
Pharmacology, University of Nigeria Nsukka. They were
housed in standard cages and allowed to acclimatize to
laboratory conditions for fourteen days, prior to the
commencement of research. The animals were fed with
rat chow and water ad libitum
Animal Feeds Formulation
Bags of pellet animal chow of the same make were
purchased from Umuahia main market Abia State Nigeria
and formulated into powdery form with the various
percentages of the samples. Sample A was formulated
with 60% sample and 40% normal rat chow while sample
B was formulated with 30% sample and 70% normal rat
chow.
Seeds of Lagenaria sphaerica (Bottle gourd) were
harvested from local farms in Umuahia North and South
Local Government Area of Abia State, Nigeria.
The seeds were peel and sundried. The sundried
samples were ground into powdery form. Sixty
grams(60g) of the sample was mixed with fourty
grams(40g) of normal rat chow (sample A) while seventy
grams(70g) of normal rats chow was mixed with thirty
grams of the sample (sample B). The formulated feed
was made into pellet form.
Experimental Design:
Thirty Six (36) male albino rats aged 8 weeks weighing
112g-118g were used for this study. The animals were
randomly assigned into three (3) groups of twelve (12)
rats in each group. Group 1 the control group was fed
normal rat chow and water.
Group II was fed sample A formulation as indicated while
group III received sample B formulation. All animals were
allowed free access to food and water ad libitum
throughout the study period. Four albino rats were
sacrificed from each group weekly for analysis after the
feeding period.
Blood Collection:
Twenty eight (28) days after feeding the rats with the

Serum lipid profile and liver enzymes of Rats fed Lageneria sphaerica (Wild bottle gourd) supplemented diet

Int. Res. J. Biochem. Biotechnol.

029

Table I. shows the mean weights of rats before and after feeding Lagneria sphearica supplemented diets (g)
GROUPS

Initial
average
weight of rats (g)

Final weights after

Feeding (g)

Mean change
weight(g)

Group I
Group II
group III

114.552.97
114.706.09
114.857.10

136.966.48
100.255.48
110.884.71

22.413.51
-14.450.61
-4.972.39

in

Results represent mean standard deviation (n=4)

Legend: Group I = Control (Fed normal rat chow)
Group II = Fed 60% sample and 40% normal rat chow
Group III = Fed 30% sample and 70% normal rat chow.

formulated feed, they were fasted overnight,

anaesthesized with chloroform and blood collected by
cardiac puncture using syringes and needles. Blood
samples from each animal were pooled into a test tube
and divided into two. The first parts were dispensed into
heparinized tubes for haematological analysis. The
second parts of the blood sample were allowed to stand
for about 15 minutes to clot and further spun using
centrifuge. Serum from these second parts of the blood
samples was pooled into sterile test tubes for the
measurement of liver enzymes and other parameters.
Histological Studies:
The method of Baker and Silverton (1985) was adopted
in the preparation of slices of previously fixed tissues
(liver) for histological examination. Following the
decalcification, dehydration, impregnation, embedding
and section cutting, the tissues were stained using the
Mayers acid-alum- haematoxylin and Eosin staining
techniques then mounted in neutral balsam. The slides
were then examined microscopically for histological
changes.
Biochemical Analyses
Serum Alanine Aminotransferase (ALT) activity and
serum Aspartate Aminotransferase (AST) activity were
determined by the colourimetric method described by
Reitman and Frankel (1957) while Alkaline phosphatase
activity of the serum was determined by the method
described by Bessey et al., (1946) using commercial
diagnostic kit (Randox, United Kingdom). Serum total
protein was determined as described by Henry et
al.,(1974) while serum triglyceride, High density
lipoprotein and Low density lipoprotein Cholesterol were
determined by the method of Fossati and Prencipe,
(1982) using Randox Reagent kit.

(ANOVA).Means
were separated using One way
analysis of variance.

RESULTS
From table I. there was an observed reduction in weight
of group II and III rats. Weight reduction was much more
pronounced in group II animals compared to group III and
control rats. Table II shows Serum lipid profiles and liver
enzymes activities of male albino rats fed Lageneria
sphaerica supplemented diet after 35 days. Results
indicates that AST, ALT and ALP activities significantly
increased (P<0.05) in group II rats compared to groups III
and control rats. Similarly triacylglycerol and total protein
in the serum were significantly higher (P>0.05) in the
group II rats than in group III and control. Although serum
total cholesterol levels remained unchanged significantly
(P<0.05) in all the test group and control, HDL level
significantly increased (P<0.05) in group II and III
compared to group I (control) animals. However
increased supplementation of the test diet led to
enhanced reduction (P<0.05) in serum LDL of the test
groups (II and III) compared to control group (I).
Interestingly all the parameters tested after 35 days of
feeding showed similar pattern after 42 and 49 days of
feeding the animals with Lageneria sphaerica
supplemented diet (Tables III and IV). Figures I- VII
show the histologies of livers from test and control
animals after 49 days (7 weeks) of feeding Lageneria
sphaerica supplemented diets. There was an enlarged
central vein (Fig II) and increased vascularity (Fig v) of
liver samples from group II animals after 49 days of
feeding test diets. Also there was evidence of fat
infiltration of liver of group II animals at the end of feeding
regimen. Similarly the group III animals liver samples
showed evidences of vascularization (Fig III) and
enlarged central vein with atropy of hepatocytes (Fig VII).

Statistical Analysis
DISCUSSION
The statistical analysis of result was done using students
package for social sciences(SPSS) and data collected
were
analyzed
using
Analysis
of
Variance

The Lageneria sphaerica supplemented diet fed to the

experimental animals led to a significant (P<0.05) loss of

Serum lipid profile and liver enzymes of Rats fed Lageneria sphaerica (Wild bottle gourd) supplemented diet

Chinyere et al.

030

Table 2. shows Serum lipid profiles and selected liver enzymes activities of male albino rats fed Lageneria
sphaerica supplemented diets after 35 days

AST (I/)
ALT (I/)
ALP (I/)
TG(mg/dl)
TP(g/dl)
TC(mg/dl)

Group I
24.00 3.46
9.00 2.00
232.63 36.52
79.40 0.87
5.15 0.84
71.82 14.04

HDL(mg/dl)

30.50 0.16

LDL(mg/dl)

25.44 0.05

Group II
31.88 3.47
12.25 2.60
519.47 95.01
92.10 2.72
9.74 0.16
64.17 8.93

35.800.08

9.950.04

Group III
27.38 7.82
9.63 1.11
424.58 68.78
84.85 1.06
8.63 0.32
64.60 8.80

35.60 0.16

12.03 0.01

Results represent mean standard deviation (n=4). Values in the same roll having the same alphabet are not
significantly different (P>0.05)
Legend: Group I = Control (Fed normal rat chow)
Group II = Fed 60% sample and 40% normal rat chow
Group III =Fed 30% sample and 70% normal rat chow

Table 3. Shows Serum lipid profile and selected liver enzymes

activities of male albino rats fed Lageneria sphaerica
supplemented diet after 42 days
Group I
AST (Iu)
ALT (I/u)
ALP (I/u)
TG (mg/dl)
TP (g/dl)
TC (mg/dl)
HDL (mg/dl)
LDL (mg/dl)

Group II
a

23.503.79
a
11.132.25
a
306.3274.91
a
81.292.27
a
6.731.27
a
101.6516.47
a
45.001.63
a
40.310.09

Group III
b

36.754.97
b
14.503.67
b
445.1046.69
b
107.331.41
b
11.390.61
b
85.5312.60
b
48.50 0.16
b
15.56 0.36

33.134.87
a
14.130.75
c
386.0827.78
97.564.08c
c
9.660.70
c
93.2113.29
a
46.00 1.63
c
27.70 0.24

Result represent mean standard deviation (n=4). Values in the same roll
having the same alphabet are not significantly different (P>0.05)
Legend:
Group I = Control (Fed normal rat chow)
Group II = Fed 60% sample and 40% normal rat chow
Group III =Fed 30% sample and 70% normal rat chow

weight after the feeding period. This loss of weight by

experimental animals increased significantly with
increased Lageneria sphaerica supplementation. These
findings are in agreement with those of Priyaharsini and
Sarojini (2000) who reported that bottle gourd induced
significant weight reductions in experimental animals.
Causes of weight reduction in experimental animals may
include toxicity of the fed diet, unacceptability of diet by
animals, indigestion and presence of non-nutritional

factors in the fed diet. However, there was an observed

necrosis of the liver of the test animals. The observed
vascular degeneration of hepatocytes was more
pronounced in group 11 animals fed 60% Lageneria
sphaerica supplemented diet. This is in contrast to group
1(control) animals fed normal rat chow which had normal
hepatocytes after the feeding period. Hanaa and Hala
(2010) reported distorted histological structures of hepatic
lobules in rats fed with bottle gourd.

Serum lipid profile and liver enzymes of Rats fed Lageneria sphaerica (Wild bottle gourd) supplemented diet

Int. Res. J. Biochem. Biotechnol.

031

Table 4. Serum lipid profile and selected liver enzymes activities of male albino rats fed Lageneria sphaerica supplemented
diet after 49 days

Group I
AST (I/u)
ALT (I/u)
ALP (I/u)
TG (mg/dl)
TP (g/dl)
TC (mg/dl)
HDL (mg/dl)
LDL (mg/dl)

Group II
a

26.635.71
a
12.252.60
a
319.3883.15
a
93.648.58
a
7.501.82
a
109.7113.29
a
45.500.16
a
45.480.02

Group III
b

37.383.42
b
17.882.25
b
483.8662.09
b
121.711.70
b
13.231.90
b
89.039.72
b
48.800.08
b
15.890.09

32.504.53
c
16.754.50
a
398.948.51
c
112.135.54
c
10.240.41
a
93.496.68
b
48.210.06
c
22.860.11

Result represent mean standard deviation (n=4). Values in the same roll having the same alphabet are not significantly different
(P>0.05)
Legend:
Group I = Control (Fed normal rat chow)
Group II =Fed 60% sample and 40% normal rat chow
Group III = Fed 30% sample and 70% normal rat chow

Figure 1. Histological picture of liver of rats in group I (Control) showing normal liver morphology

Liver enzymes such as Alkaline phosphatase (ALP),

Alanine
aminotransferase
(ALT),
Aspartate
aminotransferase (AST) are used for liver enzyme test
(Johnston, 1999; Uboh, 2004). Vutukuru et al., (2007)
reported that alanine aminotransferase and aspartate
aminotransferase play crucial roles in transamination
reaction and can be used as potential biomarkers to
indicate hepatotoxicity. Burtis and Aswood, (1996) also
reported injury to organs like liver lead to the release of

tissue- specific enzymes into the blood stream. Therefore

increased liver enzymes in serum is an indicator of
hepatocellular damage. Results obtained in this study
showed significant (P<0.05) increases in levels of liver
enzymes in serum of test animals(group 11 and 111).
The levels of these liver enzymes in serum of test
animals increased significantly (P<0.05) with increased
supplementation of Lageneria sphaerica in the test diet
compared to control animals. This observation is in line

Serum lipid profile and liver enzymes of Rats fed Lageneria sphaerica (Wild bottle gourd) supplemented diet

Chinyere et al.

032

Central vein
Figure 2. Histological section showing enlarged central vein of group II rats

Hepatocytes

Enlarged central vein

Figure 3. Histological section of group III rats showing liver tissue with distorted interstitial (stroma) and structural hepatocytes (atrophy) after the
feeding period. Observe an enlarged central vein with evidences of degeneration within the stroma The laminae and portal triads are not distinct.

with the findings of Priyadharsini and Sarojini (2000) who

opined that with toxic substances liver enzymes leak out
of the compartment into the serum. The observed

increase in alkaline phosphatase (ALP), Aspartate

aminotransferase (AST) and Alanine aminotransferase
(ALT) in test rats serum relative to control may be

Serum lipid profile and liver enzymes of Rats fed Lageneria sphaerica (Wild bottle gourd) supplemented diet

Int. Res. J. Biochem. Biotechnol.

033

Central vein
Figure 4. Shows the central vein of group II rats liver after the feeding period. Central vein enlarged.

Increased vascularity in group II animals

Figure 5. Histological section of the rat liver showing increased vascularity in group II animals. All other tissue components appear
normal.

attributed to the presence of some toxic substances in

Lageneria sphaerica which damaged hepatocytes and
caused the enzymes to leak into the serum. This
condition is similar to that observed in hepatitis condition
in humans. Many authors (Nyblom et al., 2006; Enemor
et al., 2005; Friday, 2004; Nyblom et al., 2004; Nsirim,
1999; Mathew, 2000) have noted that transaminases
(AST, ALT and ALP) leak into the circulation when the
functional integrities of liver cells are compromised.

Lipids play important roles in cardiovascular diseases,

not only by way of hyperlipidaemia and the development
of atherosclerosis, but also by modifying the composition,
structure, and stability of cellular membranes. Excess
lipids in the blood are considered to accelerate the
development of arteriosclerosis and are the major risk
factor in myocardial infarction (Mathew, 2000).
High levels of circulating cholesterol and its accumulation
in heart tissue are well associated with cardiovascular

Serum lipid profile and liver enzymes of Rats fed Lageneria sphaerica (Wild bottle gourd) supplemented diet

Chinyere et al.

034

Hepatocytes
Portal tract
Fig VI shows group I (control) rat liver .Histological picture indicating normal hepatocytes and portal tract.

damage. An altered lipid metabolism can alter the cardiac

function by changing the properties of cardiac cell
membrane and these changes may contribute to the cell
death that follows coronary artery occlusion. The cardiac
muscle generally utilizes fatty acid as the major source of
energy of the total oxygen consumption; 6090% is
utilized to oxidize fatty acid.
Accumulation of triglycerides is one of the risk factors in
Coronary Heart Disease (CHD) (Mahale et al.,2008). The
triacylglyceride results showed that group II animals had
significantly (P<0.05) higher value (92.102.72 mg/dl) in
serum when compared to group III (84.851.06mg/dl)
and control animals (79.40 0.87). The observed
significant differences in the test groups could be
attributed to the bioactive constituents of the seeds. The
nutrient composition of Lageneria sphaerica has been
presented by Chinyere et al.,(2009) as an oil seed.
Dietary fat has been strongly implicated as a factor
contributing to elevated circulating total cholesterol
(Akiyama et al., 1997). However, there was an increase
(P<0.05) in serum total protein, HDL and a decerase
(P>0.05) in serum total cholesterol in the test groups (II
and III). These are similar to results of Hanaa and Hala,
(2010). The decrease in total cholesterol could be

attributed to bioactive constituents which promoted the

actions of HDL responsible for transporting cholesterol
out of the blood. This agrees with Mohale et al., (2006)
who evaluated antilipidemic effect of Lageneria species.
Similar results was also obtained by Priyadharsini and
Sarojini(2000). Chinyere et al., (2009) reported that
Lageneria sphaerica has high fiber content. Since fiber
bind bile salts which are then excreted in the stool, the
reduction in bile salts in the enterohepatic circulation
causes the liver to increase the conversion of cholesterol
to bile acids to maintain normal bile acid pool
(Priyadharsini and Sarojini, 2000). Liu et al., (2008)
reported that low levels of saturated fatty acids and high
levels of monounsaturated and polyunsaturated fatty
acids and other bioactive molecules in plants seed
prevent hypercholesterolemia. These bioactive molecules
include plant protein, dietary fiber, phytochemicals and
lignins. Also as observed in this study and reported by
Hanna and Hala (2010), there was a significant decrease
(P<0.05) in serum LDL levels of test animals with
increased supplementation of Lageneria sphaerica
compared to control animals. This could be a beneficial
effect of consumption of wild bottle gourd seeds. This is
because low serum LDL levels signify less risk to

Serum lipid profile and liver enzymes of Rats fed Lageneria sphaerica (Wild bottle gourd) supplemented diet

Int. Res. J. Biochem. Biotechnol.

Hepatocytes

035

Enlarged central vein

Figure 7. Histological section of the group III rat liver showing liver tissue with distorted interstitial (stroma)
and structure hepatocytes (atrophy). There is an enlarged central vein (cv) with evidence of degeneration
within the stroma. The lamina and portal triads are not distinct.

coronary heart diseases. In conclusion, this study has

shown the hepatotoxic effect of Lageneria sphaerica
despite some beneficial effects obtained including high
serum HDL, low LDL and total cholesterol. Therefore
more detoxification work need to be done in order to
harness these benefits when the toxin are identified.

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Accepted 07 October, 2015.
Citation: Chinyere GC, Ikpoh JC, Osuocha KU (2015).
Serum lipid profile and liver enzymes of Rats fed
Lageneria sphaerica (Wild bottle gourd) supplemented
diet. International Research Journal of Biochemistry and
Biotechnology, 2(2): 028-036.

Copyright: 2015 Chinyere et al. This is an openaccess article distributed under the terms of the Creative
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Serum lipid profile and liver enzymes of Rats fed Lageneria sphaerica (Wild bottle gourd) supplemented diet