The anti-anaemic
activity of the ethanol extract of Abelmoschusesculentus (okra leaf) on phenyl-hydrazine induced
anaemia in female wistar rats has been
investigated. Anaemia is a known clinical condition linked with decreased hemoglobin concentrations and has high morbidity and
mortality in females. It results in poor work productivity among adults and
impairs cognitive function. The research study was targeted at determining
the anti-anaemic effect of ethanolic leaf extract
of Abelmoscusesculentus
(ELAE) on phenyl-hydrazine induced anaemic Wistar
rat. Twenty (20) female non gravid wistar rats
weighing 150-180 were used and divided into four (4) groups of 5 rats each.
The groups are namely controlled group A (n=5) feed and water only, and test
groups (B, C and D). After the induction of anaemia using phenylhydracine
and extract feed of A.esculentus
for 14 days, blood samples were collected from the animals. Haematological
tests were done using Auto-analyzer
Sysmex-xt-2000i. The result showed that LD 50 was above 5000mg/kg. Phenyl
hydrazine (PHZ) caused a significant reduction in RBC, PCV, HB, MCH and MCHC,
further RBC and PCV showed significant (P<005) increase at 300mg/kg of
ELAE and hemoglobin MCH, MCV, and MCHC were
attenuated at 150mg/kg and 300mg/kg. It could be deduced from this study that
the plant Abel Moschusesculentus
possesses active principles that interfers with haemopoietic system hence acting as an anti-anaemic
plant.
For decades,
medicinal plants are used to treat health disorders, add flavor, prevent disease
epidemics and conserve food.
Medicinal plants are
excellent resource of traditional medicines from which different modern
medicines are developed. They contain several secondary metabolic responsible
for the biological characteristics of plant species used throughout the world
(Dar, 2017). The plant okra (Abelmoschusesculentus) is the only vegetable crop that is of
significance in the Malvaceae family and is very
popular in the Indo-Pak subcontinent. Abelmoschusesculentus L
(Family: malvaceae) also known as Hibiscus esculentus
is an important vegetable, widely distributed from Africa to Asia, Southern
Europe and America that is more commonly known as Ladies finger, okra or gumbo.
However, the most significant nutritional aspect of the plant is the dried
seeds; the oils from it are edible and the residual meal after oil extraction
is rich in protein. (Doreddula,
2014, chowdhury, 2019).
The lady’s finger is
used in the management of depression and weakness, with effectiveness in ulcer
and joint healthiness. The fibers of ladies finger help to stabilize blood
sugar by regulating the route at which sugar is absorbed from the intestinal
tract and has proven to contain polysaccharides that possess hepatoprotective activities (Hu, 2014).
Anaemia is associated with
various physiological consequences including impaired tissue, oxygen delivery,
weakness, fatique and loss of productivity from
reduced work capacity, cognitive impairment and increased susceptibility
infections (Jamnok, 2020) and imposes a substantial
economic burden on an individual. In almost all the developing countries,
between one third and one-half of the females and children are anemic. The use
of medicinal plants in the management of anaemic
conditions has been on the rise in Nigeria and around the globe.
Medicinal
values of A. esculentus
Extracts of Abelmoschusesculentus
contain distinct nutrients and phytochemicals that are significant for its
antioxidant, anti-inflammatory and immunomodulatory, anti bacterial, anti cancer, anti diabetic, organ protective and neuro
pharmacological activities. Moreover, the plant also has lipid-lowering trypsin
inhibitory, haemagglutinating, anti-adhensive and anti-fatigue (Durazzo,
2019). Reports have shown that pheny/hydrazine is known as a hemolytic agent in the cause
of anaemia in experimental model.
Nutritional
and phytochemical content:
Abelmoschusesculentus leaves per 100gm contains energy (33 kcal),
carbohydrates (7.45gm). sugar (1.48gm), dietary fibers (3.2gm) fat (0.19gm),
protein (2gm), water (90.19gm), vitamin A 36µg (7%), thiamin 0.2mg (17%)
riboflavin 0.006mg (5%). Niacin 1.0mg (7%), vitamin C 23mg (28%), vitamin K
31.3µg, (30%), calcium 82mg (8%), iron 0.62mg, (5%), magnesium 57mg (16%),
potassium 299mg (6%), and zinc0.5mg
(6%) (Chowdhury et al, 2019).
The plant is rich in
distinct phytochemicals such as hyperbole/hyperin, flavenoid, glycoside coumarinscopoletin, uridine.
Islam (2019) reported that the petals are rich in protein, fat, minerals
(potassium, sodium, magnesium, sulphur, copper,
manganese and iodine) carbohydrate, calcium and phosphorus.
The use of medicinal
plants in the management of anaemic conditions has
been on the rise in Nigeria and around the globe. Our curiosity is aimed at
investigating the anti-anaemic activity of the ethanolic extract of Abelmoschusesculentus leaf on phenyl-hydrazine
induced anaemicWistar rat.
MATERIALS
AND METHODS
Animals:
Twenty (20) non
gravid female Wistar rats weighing 150-180g were
selected and used in this study. They were housed in animal house in the
Department of Physiology, chukwuemekaOdumegwuUniversity, Uli
Campus, Anambra State. The animals were kept in
standard cages at a room temperature of 25±20C. They were maintained
with standard laboratory chow (Grower feed) and water ad libitum. They were acclimatized
for a period of two weeks before the administration of phenyl-hydrazine for the
induction of anaemia, and ethanolic
leave extract of Abelmoschusesculentus
leaves, that lasted for 14 days.The
animals were kept on 12 hours light and dark cycles. However, the study was
conducted in accordance with the recommendation from the declaration of
Helsinki on guiding principles in care and use of animals.
Experimental
Designs:
The experimental
animals were randomly distributed into four (4) groups of 5 rats per cage.
Group A served as the control groups. They received feed and distilled water ad
libitum. Group B received 40mg/kg of phenyl hydrazine (PHZ) only. Group C
received 40mg/kg of phenyl hydrazine and treated with 150mg/kg of ehtanolic leave extract of Abelmoschusesculentus.
Group D received
40mg/kg of phenyl hydrazine and treated with 300mg/kg ethanolic
leave extract of Abelmoschusesculentum.
The median lethal
dose (LD 50) of ethanolic leave extract of Abelmoschusesculentus in
rats was determined using the method of Dietrich Lerke,
1983. The procedure is by increasing the concentration of the extract given to
the rat (per body weight) in each group consisting of eight (8) rats per groups
for five (5) days. The concentration used are 500mg/kg,
1000mg/kg, 1,500mg/kg, 6000mg/kg and 7000 mg/kg. the
mortality rate was determined and a graph plotted to determine the Lethal Dose
(LD 50).
Plant
collection and preparation of extract of Abelmoschusesculentus.
Samples of Abelmoschusesculentus
leaves were harvested from a local farm in NnewiAnambra State. They were washed in running tap water to
remove dirt and air dried under ambient temperature. The dried leaves of Abelmoschusesculentus
was milled into a coarsely powered form using a local grinder 250g of the dried
leaves was macerated in 1000mls of 95% absolute ethanol for 48hrs. It was
filtered using a clean handkerchief and further filtration usingwhatman’s
No. 1 filter paper. The filtrate was concentrated using a rotator evaporator
and dried further using a laboratory oven at 450C into a gel-like
form. The extract were preserved in air tight container and kept in a
refrigerator for further usage. The extraction method was done with
modifications as described by the method of Al-Attar and Abu Zeid, (2013).
Phytochemical
Analysis
The procedure for
phytochemical analysis of the elements present in A. esculentus is as described by Harbourne (1973). Trease and Evans (1996). The extract was tested for alkaloids, flavenoid, calcium, carbohydrates, steroids, fat and oil,
reducing sugars, saponin, glycosides, resins and
acidic compounds.
Induction of Anaemia using Phenyl-hydrazin
Anemia was induced in the experimental rats in groups B, C and D using phenyl hydrazine
by dissolving 1.0ml phenyl hydrazine in 5.4ml of Normal saline. The solution
was injected into the 40mg/kg body weight for two consecutive days as described
by Gheith and El-Mahmoudy
(2018) with modification.
Collection
of Blood samples:
The animals were anaesthesized with chloroform in an enclosed chamber after
24 hours of the last administered dose of the ethanolic
leaf extract of Abelmoschusesculentus;
ocular puncture was done according to the Parasuraman
et al, 2010. The blood was obtained through the orbital sinus using a hepalinized capillary tube. Samples were stored in EDTA
containers and kept in the refrigerator.
Haematological Tests:
Haemoglobin concentration, Red
blood cell count, packed cell volume, mean corpuscular volume, mean cell haemoglobin concentration were done using an automated
blood analyzer sysmex X T-2000i as described by the
method of Gheith and El-Mahmoudy,
2018
Statistical
analysis:
The data obtained
from the research were analyzed using SPSS version 25. All the haematological parameters done were analyzed using ANOVA
followed with Post HOC multiple LSD. Values were presented as Mean and SEM, and
considered significant at P<0.05.
RESULTS
Table
1: Effect of ethanolic leave extract of Abelmoschusesculentus on
RBC haemoglen and PCV of Phenyl hydrazine
Groups
Haemoglobin Concentration (Mean±SEMP)
Packed cell volume (mean± SEMP)
RBC volume (Meant± SEMP)
Group
A
n=5,
control
14.9±
0.42
45.8±1.0
6.75±0.750
Group
B
N=5, phenyl
hydrazine only for 14 days
10.0±0.49
35.1±0.4
4.17±0.34
Group
C
N=5,
phenyl hydrazine and 150mg/kg of A. esculentus
13.4±1.86
41.8±5.7
5.43±0.860
Group
D
n=5,
phenyl hydrazine and 300mg/kg of A. esculentus
16.7±0.3
48.1
± 0.7
6.46±
0.380
P-value
P<0.05
P<0.05
P<0.05
Table 2: Effect of Ethanolic
Leave extract of Abelmeschusesculentus on
MCV, MCH and MCHC of Pheny/hydrazine induced anemia.
Groups
MCV (Mean±SEMP)
MCH (mean± SEMP)
MCHC
(Meant± SEMP)
Group A
n= 5 Control
61.7± 1.5
19.1±0.1
31.3±0.8
Group B
n=5
Pheny/drazine only for 14 days
63.4+4.1
15.8± 0.2
25.9 ±1.2
Group C
n=5
pheny/drazine only for 14 days
63.4± 4.1
15.8±0.2
25.9±1.2
Group C
n=5
pheny/hydrazine and
150mg/kg of A. esculentus
for 14 days
86.2±0.9
25.4±0.5
30.8±1.4
Group D n=5
Pheny/drazine and 300mg/kg of A. esculentus
95.8±0.4
28.6±0.6
31.1±2.1
P-value
P<0.05
P<0.05
P<0.05
Table
3: Phytochemical studies on the elements of Abelmoschusesculentus
Constituents of A. esculentus
Calcium phosphorus
Carbohydrate
Flavenoids glycoside fats and
oil
Resins steroids tanminssapomis
Degree of
concentration
++
+++
+
-
Key: - (Negative), +
(present in small quantity)
++ (present in
moderate quantity), +++(present in large quantity)
Fig 1:
Lethality study (LD 50) of the effects of administering graded doses of
(500-7,000mg per kg 1.p rat) Abelmoschusesculentus against the
percentage mortality.
Graded doses
0.5-7.0mg/kg
Lethality dose (LD
50) of A. esculentus is above 5000mg
DISCUSSION
The anti-anemic
activity of the ethanolic extract of Abelmoschusesculentus on
pheny/hydrazine, induced anemia in female wister rats has been studied.
The phytochemical
analysis of the plant shows that A. esculentus contains plant elements (Table 3) such as
calcium, carbohydrates phosphorus tannins, saponins, flavenoids and terpenes
The study revealed a
significant decrease in red blood cell count (Table 1) in group B compared to
Group A; however a non–significant(p> 0.05) increase was revealed in the
mean RBC count in group C and a significant (P<0.05) increase in group D
compared to group B. The hemoglobin concentration result showed a significant
(P<0.05) increase in group D compared to group B. The hemoglobin
concentration result showed a significant (P<0.05) groups C and D had a
significant increase (P<0.05) in hemoglobin level compared to group B. Packed
cell volume showed a significant (P>0.05) decrease in group B compared to
group A; however a non-significant (P>0.05) increase was revealed in group C
anda
significant (P<0.05) increase in group D compared to group B. The mechanism
of action of the reduction in RBC, PCV and haemoglobin
concentrations following pheny/hydrazine (PHZ)
induction is attributed to the presence of Lipid peroxidation of the RBC, which
yields formation of Methaemoplobin and later converts
into irreversible hemi-chromes which causes precipitation of hemoglobin in the
formation of Heinz bodies (Sani, 2019).
Gupta, (2017) reported
a reduction in RBC, hemoglobin, and PCV, following PHZ inductionof hemolytic anemia which has
corresponds with this study report.
Toma et al, (2015) and Onyeabo, 2018 reported a significant decrease in RBC, PCV,
and haemoglobin level following anaemia
induced toxicity caused by PHZ. However, administration of ELAE showed
attenuation of the RBC, PCV and haemoglobin level
based on dose, dependent, revealing increase in these parameters at stem
levels. The report of Toma et al, (2015) has similar
result with our study which showed attenuation of RBC, PCV and haemoglobin level near normalcy following administration of
pumpkin extract. Onyeabo 2018 has similar report to
this study, which revealed attenuation of the RBC, PCV and hemoglobin
concentration.
The result of the lethality
study (LD50) shows that the LD50 in rats using Abelmoschousesculentus was
15> 5,000 mg/kg (fig 1) hence the 150mg/kg (Grp c)
and 300mg/kg (Grp D) concentration of extract used in
this research were safe throughout the period of study
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Cite this Article: Oguwike, RC; Nwogbo, MU; Okonkwo,
OB; Oguwike, FN; Oguwike,
BC (2023). The Anti-Anaemic Activity of the Ethanolic Extract of Abelmoschusesculentus (Okro
Leaf) on Phenyl Hydrazine Induced Anaemia in Female
Wister. Greener Journal of Epidemiology
and Public Health, 11(1): 29-34.
