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Effects of aqueous extract of Basella alba leaves on haematological and biochemical parameters in albinorats

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The effects of the aqueous leaf extract of Basella alba on haematological and biochemical parameters were studied in Wistar strain albino rats. Twenty four (24) Wistar strain albino rats were randomly distributed into four groups of six (6) rats each. Group I rats served as control and received 10 ml/kg of normal saline, while group II, III and IV received 60, 80 and 100 mg/kg of aqueous leaf extract of B. alba, respectively, for two weeks. Administration of the extract was done orally. At the end of the treatment period, haematological parameters (red blood cell count, white blood cell count, platelet count, packed cell volume and haemoglobin concentration) and biochemical parameters (alkaline phosphatase (ALP), alanine aminotransaminase (ALT) and aspartate aminotransaminase (AST)) were determined. The results showed that B. alba significantly increased (p < 0.05, 0.01) red blood cell count, white blood cell count, packed cell volume, haemoglobin concentration and platelet count. However, the extract significantly (p < 0.05, 0.01) reduced the activity of the liver enzymes such as ALP, ALT and AST. The decreases were dose dependent. In conclusion, adding B. alba leaves as part of daily diet may reduce anaemia and maintain good health.
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African Journal of Biotechnology Vol. 9(41), pp. 6952-6955, 11 October, 2010
Available online at http://www.academicjournals.org/AJB
ISSN 1684–5315 ©2010 Academic Journals
Full Length Research Paper
Effects of aqueous extract of Basella alba leaves on
haematological and biochemical parameters in albino
rats
Bamidele, O.
1
*, Akinnuga, A. M.
2
, Olorunfemi, J. O.
3
, Odetola, O. A.
3
, Oparaji, C. K.
3
and
Ezeigbo, N.
3
1
Department of Physiology, Bowen University, Iwo, Osun State, Nigeria.
2
Department of Nursing Science, Lead City University, Ibadan, Oyo State, Nigeria.
3
Department of Physiology, Madonna University, Okija, Anambra State, Nigeria.
Accepted 2 September, 2010
The effects of the aqueous leaf extract of Basella alba on haematological and biochemical parameters
were studied in Wistar strain albino rats. Twenty four (24) Wistar strain albino rats were randomly
distributed into four groups of six (6) rats each. Group I rats served as control and received 10 ml/kg of
normal saline, while group II, III and IV received 60, 80 and 100 mg/kg of aqueous leaf extract of B. alba,
respectively, for two weeks. Administration of the extract was done orally. At the end of the treatment
period, haematological parameters (red blood cell count, white blood cell count, platelet count, packed
cell volume and haemoglobin concentration) and biochemical parameters (alkaline phosphatase (ALP),
alanine aminotransaminase (ALT) and aspartate aminotransaminase (AST)) were determined. The
results showed that B. alba significantly increased (p < 0.05, 0.01) red blood cell count, white blood cell
count, packed cell volume, haemoglobin concentration and platelet count. However, the extract
significantly (p < 0.05, 0.01) reduced the activity of the liver enzymes such as ALP, ALT and AST. The
decreases were dose dependent. In conclusion, adding B. alba leaves as part of daily diet may reduce
anaemia and maintain good health.
Key words: Basella alba, haematological parameters, biochemical parameters, albino rats.
INTRODUCTION
Basella alba (family Basellaceae) is a fast growing
vegetable, native to tropical Asia, probably originating
from India or Indonesia and extremely heat tolerant
(Grubben and Denton, 2004). It is grown throughout the
tropics as a perennial and in warmer temperate region as
an annual crop. Its thick semi-succulent heart-shaped
*Corresponding author. E-mail: femi_bayo_is@yahoo.co.uk.
Tel: +2348067463528.
Abbreviations: ALP, Alkaline phosphatase; ALT, alanine
aminotransaminase; AST, aspartate aminotrasaminase; PCV,
packed cell volume; RBC, red blood cell; count, WBC, white
blood cell; Hb, haemoglobin; EDTA, ethylenediaminetetraacetic
acid.
leaves have a mild flavour and mucilaginous texture. It is
commonly known as Malabar, Ceylon, East-Indian, Surinam
and Chinese spinach (Facciola, 1990).
It is high in vitamin A, vitamin C, vitamin B9 (folic acid),
calcium, magnesium and several vital anti-oxidants. It is
low in calories by volume and high in protein per calorie
(Duke and Ayensu, 1985). A work done in Bangladeshi
showed that the daily consumption of Indian spinach has
a positive effect on vitamin A stores in populations at high
risk of vitamin A deficiency (Haskell et al., 2004). In
addition, the cooked roots and leaves have been reported
to be used in the treatment of diarrhoea and as laxative,
respectively (Larkcom, 1991; Philips and Rix, 1995).The
flowers are used as an antidote for poisons (Duke and
Ayensu 1985). It is also a safe aperient for pregnant
women and its decoction has been used to alleviate
labour (Duke et al., 1985). Moreover, it is locally reported
to be used in the treatment of anaemia. Its extract has
demonstrated androgenic potential in adult rats and bull
Leydig cells (Moundipa et al., 2005).
In spite of its many biological uses and the fact that
several studies (Alada et al., 2004; Bolarinwa et al., 1991)
have implicated the relative importance of some animal
and vegetable protein diets in the formation and com-
position of blood, there is dearth of information encountered
in the literature on the effect of B. alba on haematological
parameters. Also, in view of the increasing use of B. alba
in several communities by traditional healers to cure
many diseases in patients without considering its adverse
effects, it is necessary to scientifically investigate its
effect on biochemical parameters. Therefore, the present
study was undertaken to investigate the effect of aqueous
extract of B. alba leaves on haematological and biochem-
ical parameters in Wistar strain albino rats.
MATERIALS AND METHODS
Animal model
Wistar strain albino rats weighing between 140 180 g were used.
The rats were purchased at the animal house of the College of
Medical Sciences, University of Nigeria, Nsukka. The rats were
housed in wire mesh cages under standard conditions (temperature,
25 - 29°C, 12 h light and 12 h dark cycle) and fed with standard rat
pelleted diet and water was given ad libtum.
Plant materials
The fresh leaves of B. alba (Indian spinach) were procured from
Otor market in Lagos, western Nigeria. The plant materials were
identified and authenticated in the Department of Pharmacognosy,
Faculty of Pharmacy, Madonna University, Elele Campus. The
leaves were washed in tap water and shade-dried after which they
were reduced into fine powder by grinding. 100 g of the powered
leaves was stirred into 1000 ml of boiling distilled water. Boiling was
allowed to continue for 5 min. The mixture was kept off the hot plate,
for 30 min to allow it to infuse. It was then filtered using cheese
cloth. The filtrate was then concentrated using a rotary vacuum
evaporator to obtain the solid mass. The extract was then dissolved
in normal saline and used for the study.
Experimental design
Twenty four (24) Wistar strain albino rats were randomly distributed
into four groups of six (6) animals per group. Group I consists of
rats which received 10 ml/kg normal saline and served as the
control. Group II, III and IV received the aqueous extract of B. alba
leaves at doses of 60, 80 and 100 mg/kg, respectively.
Administration of the extract was done through gastric intubation
once a day for a period of 14 days. Blood samples were collected
from the animal through cardiac puncture into ethylenediami-
netetraacetic acid (EDTA) bottles after anaesthetizing the animals
with chloroform at the 14th day of the experiment. The blood
samples were divided into two: First portion was used to determine
haematological parameter, while the second portion was used to
determine biochemical parameters. The second portion of blood
was allowed to clot and then centrifuged at 150 g for 10 min. Serum
obtained was used for the assay of alkaline phosphatase (ALP),
alanine aminotransaminase (ALT) and aspartate aminotrasaminase
Bamidele et al. 6953
(AST).
Blood analysis
The blood samples were analyzed to determine the haematological
parameters such as: Packed cell volume (PCV), red blood cell
(RBC) count, white blood cell (WBC) count, platelet count and
haemoglobin concentration (Hb Conc.) using an automated
haematology ANALYZER KX-21N, made by sysmex Japan. The
sysmex KX-21 is an automatic multi-pair blood cell counter for in
vitro diagnostic use in clinical laboratory. It performs speedy and
accurate analysis of blood parameters and detects the abnormal
samples. The automated haematology analyzer reading correlated
well with readings by the standard manual methods (Samuel et al.,
2010). Biochemical analysis of the serum enzymes for ALT and
AST was by the method of Reitman and Frankel (1957). ALP was
assayed according to the method of Rec (1972).
Statistical analysis
All data were presented as mean ± SEM. The one way ANOVA was
used to analyze the data, followed by a post-hoc test (LSD). The
results were considered significant at p values of less than 0.05.
RESULTS
Haematological parameters
The results of the effects of the aqueous extract of B.
alba on red blood cell count, white blood cell count,
packed cell volume, haemoglobin concentration and
platelet count are shown in Table 1. Oral administration
of the aqueous extract of B. alba for two weeks caused
gradual but significant increases in the mean RBC count
in the treated groups of rats (5.87 ± 0.04, 6.24 ± 0.34 and
8.60 ± 0.28 x 10
6
cells/mm
3
for groups II, III and IV,
respectively). The increases were dose dependent. There
were increases with no significant differences in the
mean total WBC count in groups II (6.89 ± 0.65 x 10
6
cells/mm
3
) and III (7.60 ± 0.13 x 10
6
cells/mm
3
) when
compared with the control (6.70 ± 0.50 x 10
6
cells/mm
3
),
whereas there was significant increase (p < 0.05) in
group IV (8.93 ± 0.26 x 10
6
cells/mm
3
). The mean PCV
increased significantly (p < 0.05) in the treated rats as
compared with the control group. The percentage
increases were 7, 29 and 46, percent in group II, III and
IV, respectively. The mean Hb concentration also
increased significantly (p < 0.05) in a dose dependent
fashion in the treated rats when compared with the
control group. The percentage increases were 19, 22 and
50% in group II, III and IV, respectively. The mean
platelet count in group II was not significantly different
from the control group but group III and IV showed signifi-
cant increase (p < 0.05).
Biochemical parameters
The activities of the three major marker enzymes ALP,
6954 Afr. J. Biotechnol.
Table 1. Effect of aqueous extract of B. Alba leaves on some haematological parameters.
Blood parameter
Group I
(Control)
Group II
(60 mg/kg)
Group III
(80 mg/kg)
Group IV
(10 0mg/kg)
RBC (×10
6
/mm
3
)
4.61±0.06 5.87±0.041 6.24±0.34* 8.60±0.28**
WBC (×10
3
/mm
3
)
6.70±0.50 6.89±0.65 7.60±0.13 8.93±0.26*
PCV (%) 34.90±0.25 37.32±0.51* 45.10±0.38** 50.93±0.79**
Hb(g/dl) 7.80±0.25 9.30±0.51* 9.51±0.14* 11.70±0.47**
PLC (×10
3
/mm
3
)
199.87±2.13 198.56±2.16 205.45±0.10** 204.20±0.09*
Values are expressed as mean ± S.E.M. *P < 0.05, **P < 0.01 (n = 6).
Table 2. Effect of aqueous extract of B. alba leaves on some biochemical parameters.
Biochemical
parameter (µ
µµ
µ/l)
Group I
(Control)
Group II
(60 mg/kg)
Group III
(80 mg/kg)
Group IV
(100 mg/kg)
ALAT 57.50±0.44 49.80±0.32** 52.10±0.19** 39.70±0.12**
ASAT 60.65±0.85 62.79±0.52 57.56±0.36* 41.08±0.69**
ALP 400.30±0.63 345.10±0.45** 322.40±0.72** 308.58±0.58**
Values are expressed as mean ± S.E.M. *P < 0.05, **P < 0.01 (n = 6).
ALAT = Alanine aminotransaminase; ASAT = aspartate aminotrasaminase; ALP = alkaline phosphatase.
ALT and AST were markedly affected after pre-treatment
of the rats with aqueous extract of B. alba at different
doses of 60, 80 and 100 mg/kg body weight. ALT activity
significantly decreased (p < 0.01) in the rats pre-treated
with various doses of the extract as shown in Table 2.
There was a gradual decrease in the activity (62.79 ±
0.52, 57.56 ± 0.36 and 41.08 ± 0.69 µ/l) of ALT in all the
rats pre-treated with different doses of the extract.
Groups that received 80 and 100 mg/kg body weight
showed significant decrease (p < 0.05) when compared
with the control value. The activity of ALP decreased
significantly (p < 0.01) at different doses of the extract in
the pre-treated rats (345.10 ± 0.45, 322.40 ± 0.72 and
308.58 ± 0.58µ/l) as compared with the control value
(400.30 ± 0.63 µ/l). The decrease in the activity of ALP in
the pre-treated rats was dose dependent.
DISCUSSION
The results of the study shows that, the leaf extract of B.
alba administered at the dosages used and for the period
of the experiment has positive effect on the haemopoietic
system. There was increase in the haematological para-
meters (RBC, WBC, PCV, Hb Conc. and platelet) in the
treated rats when compared with the control group.
The significant increase in the haematological indices
observed following treatment with B. alba aqueous leaf
extract might not be unrelated to its chemical composition.
The chemical composition of the leaf extract include:
Proteins, fat, vitamin A, vitamin C, vitamin E, vitamin K,
vitamin B9 (folic acid), riboflavin, niacin, thiamine and
minerals such as calcium, magnesium and iron (Duke
and Ayensu, 1985; Grubben and Denton, 2004). Most of
these vitamins and minerals are well-known hematinics
and are necessary for the formation of blood cells (Ganong,
2005; Alada et al., 2004; Mitchell et al., 1976). The
observed increases in the haemoglobin concentrations
and packed cell volume in the rats treated with the
aqueous leaf extract of B. alba in this study is consistent
with earlier reports that protein-rich diets increase both
packed cell volume and haemoglobin concentrations in
human and animal studies (Alada et al., 2004; Alada, 2000;
Bolarinwa et al., 1991, Mitchell,1966).
The aqueous leaf extract of B. alba significantly
reduced the activity of the liver enzymes (ALT, AST and
ALP) in the treated rats when compared with the control
group. The effect was dose dependent. Liver enzymes
(ALT and AST) are released into the blood whenever liver
cells are damaged and enzyme activity in the plasma is
increased (Edwards et al., 1995). The fact that the
enzyme activities were reduced showed that the extract
improves hepatic functions.
Moreover, B. alba leaves are reported to contain antio-
xidant properties (Duke and Ayensu, 1985). Antioxidants
are effective scavengers of super oxide anions. They are
associated with several health benefits including their
ability to protect against oxidative damage (Olmendilla et
al., 1997). Therefore, the extract may have exhibited
hepatoprotective activity due to its antioxidant properties
which is attributable to flavonoids and carotinoids.
In conclusion, the results of this study confirmed the
use of the B. alba leaves in traditional medicine for the
treatment of anaemia. Thus, the leaves of the plant might
have a promising role in the treatment and/or prevention
of anaemia. On the other hand, the use of the leaves of
the plant at the dosages in this study may not have
deleterious effect on the body system owing to its
hepatoprotective potentials due to the presence of
antioxidants.
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