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American Journal of  Medical 
Science and Innovation (AJMSI) 

Effect of  Aqueous Stem Bark Extract of  Parkia BiglobosaParkia Biglobosa  on the Histological 
Morphology of  Liver of  Adult Wistar Rats

D. F. Ibrahim1*, A. S. Hassan1, A. Sani2, A. Zakariyya1, F. B. Shema1, R. U. Zubair1, G. Y. Riruwai1 

Volume 2 Issue 1, Year 2023
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Article Information ABSTRACT

Received: December 30, 2022

Accepted: January 19, 2023

Published: February 17, 2023

People in Africa and other developing countries depend on medicinal plants. Extract of  
Parkia biglobosa stem bark is used in Nigerian traditional medicine to treat malaria, diarrhea 
and pains. This study aimed at determining the effect of  aqueous stem bark extract of  
P.biglobosa on the histological morphology of  liver in animal models. A total of  28 wistar 
rats weighing between 100 – 150g were used for this study out of  which 12 were used for 
acute toxicity testing using Lorke’s method (1983), the test was carried out in two phases. 
In phase I, 9 adult wistar rats were divided into 3 groups and administered with 10mg/kg, 
100mg/kg and 1000mg/kg orally while in phase II, 3 wistar rats were assigned into 3 groups 
and administered with 1600mg/kg, 2600mg/kg and 5000mg/kg respectively and both they 
were observed for signs of  toxicity/mortality within 24hrs. The remaining 16 rats were 
divided into 4 groups; A, B, C and D. Group A served as control and groups B, C and D 
served as test groups. For 21 days, group A received normal feed and water only and then 
group B, C and D received oral doses of  1000mg/kg, 500mg/kg and 250mg/kg of  aqueous 
extract of  P.biglobosa stem bark respectively. A significant increase in the level of  ALP, ALT 
and AST was observed in comparison with the control, P< 0.005 was considered statistically 
significant. It was shown that liver sections in group A (control) presents no histological 
changes while the liver section of  animals treated with (1000mg/kg and 500mg/kg) of  
P.biglobosa  aqueous extract for 21 days revealed an inflammation, vascular degeneration and 
vacoulation respectively.  It was found that at higher doses P.biglobosa aqueous stem bark 
extract could be hepatotoxic.

Keywords
Parkia Biglobosa, Acute-
Toxicity, Wistar-Rats, Stem-
Bark, Liver, Intervention

1 Department of  Medical Laboratory Science, Faculty of  Allied Health Sciences, Bayero University, Kano, Nigeria
2 Aminu Kano Teaching Hospital , Kano, Nigeria
* Corresponding author’s e-mail: dahirufalalu63@gmail.com

INTRODUCTION
Parkia biglobosa is a scientific name of  African locust bean 
tree. In Hausa it is referred to as Dorawa, in Yoruba it 
is known as Igba Irugba and in Igbo it is called Origili. 
The Parkia tree, named after the famous Scottish botanist 
and surgeon, Mungo Park by Brown (1826) has long 
been widely recognized as an important indigenous 
multipurpose fruit tree in many countries of  the sub - 
saharan Africa. 
Parkia biglobosa is found naturally occurring in the 
following countries of  west Africa: Republic of  Benin, 
Burkina Fasso, Cameroon, Chad, Cote d’Voire, Central 
Africa Republic, Gambia, Ghana, Guinea Bissau, Kenya, 
Mali, Niger, Nigeria, Senegal, Sierra Leones, Sudan, Togo, 
Tanzania, Uganda and Zaire (Booth and Wickens, 1988) 
(Sina & Traore, 2002). 
In Nigeria, Parkia biglobosa is found in the Savannah zone 
with the bulk of  it in the Guinea Savannah. This is as a 
result of  its ecological and environmental requirements 
which are easily met in these areas (Oni, 1997). 
The distribution of  Parkia biglobosa in Nigeria covers 
Abuja, Adamawa, Bauchi, Gombe, Kaduna, Kano, 
Katsina, Kebbi, Kogi, Kwara, Nassarawa, Niger, Oyo, 
Taraba, Yobe, Plateau and Zamfara state. 
In west Africa the bark, roots, leaves, flowers, fruits and the 
seeds are commonly used in traditional medicine to treat 
a wide diversity of  complaints, internally and externally, 
sometimes in combination with other medicinal plants 

(Builders et al., 2011). The bark is the most important for 
medicinal uses, followed by the leaves. It have been used 
in Nigeria and other west African rural communities to 
treat a variety of  diseases (Abbie 1990; Shao 2002). 
The bark soaked in ethanol are also used in some 
communities for anti diarrhoeal properties and as 
an effective anti-snake venoms that protects against 
neurotoxic, haemotoxic and cytotoxic effects of  
poisonous snake (Agunu et al., 2005). 
The efficacy of  the various preparation of  Parkia biglobosa 
is widely acclaimed by Hausa communities of  northern 
Nigeria for the treatment of  diseases such as Malaria, 
diabetes mellitus and pains. 
The bark is boiled in water and taken as a decoction 
for the treatment of  malaria, inflammatory disease and 
infection to diarrhoea (Asase et al., 2005; Gronhaug et al., 
2008; and Tijjani et al., 2009).
A decoction of  the stem - bark is used as a mouth wash 
to relieve tooth ache as well as, a bath for fever (Ajaiyeoba 
2002). The bark is also used with lemon for wounds and 
ulcers. In Cote d’voire, a bark infusion is used as a tonic 
for diarrhoea and as an enema (Duker - Eshun et al., 2018; 
Agunu et al., 2005). 
Among the Hausa people of  northern Nigeria, Parkia 
biglobosa is used against bronchitis, Pneumonia, diarrhea, 
vomiting, sores and Ulcers.
The water extract demonstrated increase in the 
triglyceride and cholesterol level. Phytochemicals 

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generally are regarded as research compounds. Some 
phytochemicals have been used as poisons and others as 
traditional medicine (James et al., 2007). Parkia plants have 
been identified as source of  tannins, saponins, steroids, 
Reducing sugars and Glycosides.
There is increased research in to phytochemical for 
the effective therapeutic combat of  this menace. The 
therapeutic effects of  plant - based drugs have been 
documented to be due to the phytochemicals that 
constitute the plants (Kumar et al., 2012). 

MATERIALS AND METHOD
Research Design
The study was randomized control trial (an experimental 
study) and the ethical approval for the research was 
obtained from the Research ethical committee, College 
of  Health sciences, Bayero university Kano, Kano state. 

Preparation of  Experimental Animals
Twenty-eight (28) healthy Adult wistar rats of  both sexes 
weighing (100 - 150g) were purchased from the Animal 
House section of  the Department of  Pharmacology, 
Faculty of  Pharmaceutical Sciences, Bayero University, 
Kano. The animals were maintained in standard animal 
cage at the Pharmacology Department and also they  were 
acclimatized for 2 weeks prior to the commencement of  
the study. The animals were allowed to have free access 
to drinking water and standard livestock feed, also the 
animals were maintained under standard condition of  
humidity and temperature. 
A standard protocol were adhered in accordance with 
the Good Laboratory Practice (GLP). The principle of  
Laboratory animals care were also be followed in this study. 

Preparation of  Aqueous Extraction of  Parkia Biglobosa
The stem bark of  Parkia biglobosa were collected from 
Gabasawa local Government area of  Kano State. A 
quantity of  stem bark was allowed to air dried under 
shade at room temperature. Using wooden mortar and 
pestle, the dried stem bark was grinded to powder form. 
Five hundred (500) grams of  the powdered material were 
soaked in three liters (3L) of  distilled water and were 
allowed to stand for 120 hours (5 day). 
The mixture were filtered using a Whatmann’s filter paper 
to obtained the extract. The filterate were then incubated 
in water bath and evaporated under reduced pressure, the 
filterate was allowed to dried in a hot air oven at 40oc 
to give 75g of  an aqueous stem bark extract which were 
used for interventional study at different doses. 

Qualitative Determination of  Phytochemical 
Substances of  Parkia Biglobosa Aqueous Stem Bark 
Extract
A small portion of  the extract was subjected to 
phytochemical test. Methodologies for the determination 
of  Phytochemical substances used in this research were 
adapted from those reported by Keay et al. (2008) and 
Ejikeme et al. (2014)

Acute Toxicity Testing  
The LD50 of  the extract was determined using Lorke’s 
method (1983). The test were carried out in two phases;
In phase I, nine (9) wistar rats was randomly assigned into 
three (3) groups of  three (3) wistar rats each. The first 
(1st) group comprised of  wistar rats weighing 126g, 140g 
and 135g respectively and they were administered with 
10mg/kg body weight of  the extract using orogastric 
tube ( oral cannula). 
The 2nd group comprised of  wistar rats weighing 152g, 
143g and 136g respectively and they were given 100mg/
kg and the 3rd group comprised of  wistar rats weighed 
122g,131g and 138g respectively and they were given 
1000mg/kg body weight of  the extract. The animals 
was observed within 24 hours to monitor the behavioral 
changes for signs of  toxicity as well as mortality. 
In phase II, three (3) rats weighing 148g, 151 and 
153g respectively was used and randomly placed in to 
3 groups of  one (1) wistar rat each. The animals were 
administered with high doses of  1600mg/kg, 2900mg/
kg and 5000mg/kg respectively. They were then observed 
within 24 hours for signs of  toxicity and mortality. After 
24 hours, there was no any mortality in both Phase I and 
II of  the experiment.  
Therefore, 20% of  the highest dose (5000mg/kg) used 
for the LD50 was used as the highest dose (1000mg/kg) 
in this experiment. 10% of  5000mg/kg was used as the 
medium dose (500mg/kg) and 5% of  5000mg/kg was 
used as low dose (250mg/kg) in the experiment.

Animal Grouping and Intervention
Sixteen (16) rats were randomly selected and then divided 
in to Four (4) groups, labeled as  group A, B,C and D; with 
each group containing Four (4) wistar rats. The groups 
constitute of  three (3) test groups and control group. The 
animals in group A were used as the control group and 
therefore they were not received any intervention rather 
they were administered with distilled water in place of  
the intervention. The animals in group B were used as 
the first test group and were administered with high dose 
(1000mg/kg) of  the aqeous stem bark extract of  Parkia 
biglobosa orally every day for a period of  three (3) weeks. 
The animals in group C were administered with medium 
dose (500mg/kg) of  the Parkia biglobosa aqueous stem 
bark extract for a period of  three weeks. The animals in 
group D were used as another test group and they were 
administered with low dose (250mg/kg) of  the Parkia 
biglobosa aqueous stem bark extract orally for a period of  
three weeks. 
All the four (4) groups were maintained for a period of  
three weeks. 
Therefore, measurement of  the animals weight was done 
before the commencement of  the experiment, weekly 
and after administration of  the Parkia biglobosa aqueous 
stem bark extract.

Liver Function Test (LFT)
After 3 weeks of  the experiment, a blood sample were 

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obtained from orbital venous plexus, and the blood 
samples were collected in to a plain tubes, they were 
allowed to clot and centrifuged at 1500 rpm for 15 
minutes. The serum were obtained after separation and 
then used for determination of  Alanine aminotransferase 
(ALT), Aspartate aminotransferase (AST) and Alkaline 
phosphatase (ALP).

Preparation And Processing of  the Organ (Liver)  
On the Final day of  the intervention, the animals was 
sacrificed by cervical dislocation. The liver was harvested 
and then fixed in 10℅ formalin for 48 hours and label 
with tags. Gross anatomy of  the liver was studied first 
to identify any observational lesion before subjecting the 
tissue into the tissue processing protocol. The processed 
tissues were  embedded in paraffin wax and tissues 
section of  three (3) microns were cut using Leica brand 
microtome .
The histological sections floated out in tissue water bath, 
picked with a glass slide and stained with Hematoxylin 
and Eosin staining technique for demonstration of  the 
tissue architecture and examination.

Data Analysis  
Result was presented in a tabular form and 
photomicrographs were attached to the result obtained. 
All test group of  the animals was compared with the 
control group using one-way ANOVA, using the computer 
programme Statistical Package for Social Sciences (SPSS) 
software version 20, Post hoc comparison were the 
method used for the analysis. All data were expressed as 
mean ± standard error of  mean  (SEM) and P<0.005 was 
considered statistically significant.

RESULTS
Physical Property of  Stem Bark Extract of  Parkia 
biglobosa
The aqueous extract was obtained by dissolving 500g 
of  the Parkia biglobosa stem bark powder in to 3Liters 
of  distilled water. The extraction procedure yielded 75g 
of  the extract and the percentage yield was calculated as 
15% while physical properties of  the extracts are cocoa-
brownish in colour,  crystalline shiny in consistency with 
a sweet smell.

Table 1: Physical properties of  Parkia biglobosa stem bark extract
Plant part Extract type % Yield Texture Colour Smell
Stem bark Aqueous extract 15% Crystalline Cocoa- Sweet

Shiny Brownish Smell

Phytochemical Analysis
The Phytochemical screening of  Parkia boglobosa 
aqueous stem bark extract were carried out to determine 
the presence of  the following Compounds: Tannin, 
Saponin, Reducing sugars, Steroids and Phenols using a 
standard procedure reported by Keay et al., (2008) and 
Ejikeme et al., (2014). 

Table 2: Phytochemical screening of  the Aqueous stem 
bark extract of  Parkia biglobosa
Phyto-constituents Test method Result
Tannin Aqueous extract +
Saponin Ferric chloride test +
Flavonoid Ammonium 

hydroxide method
-

Cardiac glycoside Keller Killiani’s test -
Reducing sugars Ferric chloride test +
Steroids Salkowski’s test +
Terpenoids Salkowski’s test -
Phenol Ferric chloride test +
Note: ‘+’ represents Phytoconstituent present, ‘-’ represents 
Phytoconstituent absent.

Acute Toxicity Evaluation
The aqueous extract administered to the wistar rats 
showed no sign of  toxicity or behavioral changes. After 
24 hours observation, no death were recorded in both 

phase I and phase II of  the experiment. Therefore, the 
Lethal Dose ( LD50 ) of  the aqueous stem bark extract 
of  Parkia biglobosa was greater than 5000mg/kg body 
weight as described in Table 3. 

Table 3: Lethal Dose (LD50) of  Aqueous stem bark 
extract of  Parkia biglobosa in wistar rats
Dose (mg/kg) Sign of  Toxicity/Mortality

Result
Phase I Phase II

10 0/3 -
100 0/3 -
1000 0/3 -
1600 - 0/1
2900 - 0/1
5000 - 0/1
The LD50 of  the Aqueous stem bark extract was determined 
using Lorke’s method (1983) 

Effect of  Aqueous Stem Bark Extract of  Parkia 
Biglobosa on Weight of  the Experimental  Animals 
Before (Pre) and After (Post) Interventional Study 
According to the Doses Administered
During the interventional study, the weight of  the animals 
was recorded before and after the study. The mean of  
the weights of  the animals before intervention of  all the 
groups were found to be 105.50 for group A (control), 

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135.25 for group B, 122.50 for group C and 131.00 for 
group D.
These result clearly indicates that there are increase 
in mean of  the weights from group A to B before 
intervention. The mean of  the weights of  the animals 
after intervention of  group A to D were found to be 

119.25 for group A (control), 163.00 for group B, 129.00 
for group C and 174.00 for group D. 
The body mean weight difference between the groups are; 
13.75 for group A, 27.75 for group B, 6.50 for group C 
and 43.00 for group D. These result clearly indicates that 
there is slightly increase in body weight across all groups.

Table 4: Effect of  aqueous stem bark extract of  Parkia biglobosa on weight (Mean ± SEM) of  the wistar rats before 
(pre) and after (post) interventional study according to the doses administered

Pre- Weight Post-Weight
Group A 105.5 ± 3.57 119.25 ± 7.79     
Group A 135.25 ± 15.48                      163.0 ± 21.79      
Group A 122.5 ± 12.20                         129.0 ± 5.85 
Group A 131.0 ± 8.91                           174.0 ± 10.00     
Group A = Control Group, Groupn B = High dose of  aqueous stem bark extract of  Parkia biglobosa (1000 mg/kg), Group C 
= Medium dose of  aqueous stem bark extract of  Parkia biglobosa (500 mg/kg) and Group D = Low dose of  aqueous stem bark 
extract of  Parkia biglobosa (250 mg/kg).

Liver Function Test ( Liver Enzymes)

Table 5: Comparison of  liver enzymes (Mean ± SEM) among different test groups and control group
Parameter Group A Group B Group C Group C Group D
ALP (IU/L) 48.00 ± 3.24  90.00 ± 2.08  67.00 ± 3.05 53.50 ± 4.50    0.000     
ALT (IU/L) 23.25 ± 1.65 46.66 ± 2.33 41.00 ± 1.15 27.50 ± 1.50 0.000     
AST (IU/L) 23.75 ± 2.95 45.66 ± 2.02 38.33 ± 4.33 30.00 ± 3.00 0.001   
The result were expressed as Mean ± SEM and  P < 0.005  is statistically significant

Histological finding on liver of  group A (control) 
and test groups (Group B, C and D)
The liver sections of  control group (Group A) shows 
no observable changes with hepatocytes radiating from 
a distinct central vein. None of  the tissue sections from 
this group showed any histomorphological changes 
(Figure A). Group B shows area of  inflammation and 
vascular degeneration (Figure B). Group C shows an 
areas of  inflammation, degeneration and vacuolation of  
hepatocytes (Figure C). Group D shows no observable 
changes indicating that there is no evidence of  liver cell 
damage (Figure D).

Figure 1: A Photomicrograph of  liver section from group 
A (control) which shows normal hepatic portal vein

Figure 2: A Photomicrograph of  liver section from 
group B (1000mg/kg) showing areas of  inflammation 
and vascular congestion obtained using H&E Staining 
technique (mag×100)vein

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Figure 3: A Photomicrograph of  liver section from 
group C (500mg/kg) which shows areas of  inflammation 
and Hepatic portal vein obtained using H&E Staining 
technique (mag×100) vein

Figure 4: A Photomicrograph of  liver section from 
group C (500mg/kg) which shows areas of  inflammation 
and Hepatic portal vein obtained using H&E Staining 
technique (mag×100) vein normal hepatic portal vein  
obtained using H&E Staining technique(mag×100)

Figure 5: Photomicrograph of  stem bark of  Parkia 
biglobosa

DISCUSSION
In this study, the extraction procedure yielded 75g of  the 
dried extract and the percentage yield were calculated as 
15% which were used for interventional study at various 
doses.
The acute toxicity of  P. biglobosa has been investigated to 
determine any adverse effect that may arise as a result 

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of  a short time animal exposure to the extracts within 
24 hours period. Though P.biglobosa has been used by 
Traditional medical practitioners (TMPs) without report 
of  any mortality due to toxicity, this claim has been 
authenticated by the lack of  death at oral treatment of  
over 5000 mg/kg body weight of  the extract. The results 
thus suggest that the extract of  P. biglobosa has low toxicity 
(Schorderet, 1992), since the LD50 was greater than 
5000mg/kg body weight. The low toxicity obtained may 
have been responsible for its widespread use in different 
ethnotherapeutic interventions.
Rats treated with various doses of  the extract (1000 mg/
kg, 500 mg/kg and 250mg/kg) showed a significant 
increase in body weights in relation to the control animals, 
indicating that P. biglobosa has adverse effects on the body 
weight. The water extract increases serum triglycerides 
concentration and total cholesterol level.
Ordinarily, liver cell damage is characterized by a rise in 
plasma enzymes (ALP, AST, ALT, etc). In this study, there 
is a slightly increase in liver enzymes level (AST, ALT and 
ALP) compared to the control group. The group treated 
with high and medium doses shows a slight increase 
in the level of  the enzymes and also the histological 
investigation reveals an inflammation and vascular 
degeneration, therefore P. biglobosa induces hepatocellular 
damage. 
The plant kingdom represents an enormous reservoir 
of  biologically active compounds with various chemical 
structures and protective /disease preventive properties 
(phytochemicals). These phytochemicals, often secondary 
metabolites present in smaller quantities in higher plants, 
include the alkaloids, steroids, flavonoids, terpenoids, 
tannins, and many others. The active principles of  many 
drugs found in plants are secondary metabolites (Ghani, 
1990; Dobelis, 1993). Therefore, basic phytochemical 
investigation of  these extracts for their major 
phytoconstituents is also vital. In this study, the aqueous 
stem bark extract of  P.biglobosa revealed the presence of  
tannin, Saponin, reducing sugars, phenol and steroids.
The toxic effect of  water extract of  P. biglobosa on the liver 
may be due to any one or more of  the phytochemicals 
present in the extract. Furthermore the phytochemical 
screening of  the water extract of  P. biglobosa indicated 
presence of  appreciable amount of  tannins. Study 
conducted by Yamasaki et al. (2002), Bajaj (1988), showed 
that a large intake of  tannins may cause liver damage.
In this study, it was found that in control group no 
morphological changes were identified by histopathology 
in the liver suggesting that these animals were healthy and 
the condition under which the experiment was conducted 
were proper.
The animals administered with high dose of  P.biglobosa 
stem bark extract (1000mg/kg) showed areas of  vascular 
degeneration as shown in the photomicrograph in figure 
B. The animals treated with medium dose of  P.biglobosa 
stem bark extract (500mg/kg) showed a considerable 
sign of  inflammation, degeneration and vacuolation of  
hepatocyte. as shown in the photomicrograph in figure C.

The animals administered with low dose of  water extract 
of  P.biglobosa (250mg/kg) showed no remarkable liver cell 
damaged and no any sign of  inflammation as shown in 
figure D.

CONCLUSION
This study has shown the diversity in toxicity as well 
as the chemical constituent of  the aqueous stem bark 
extract of  P. biglobosa. There is slight increase in serum 
liver enzymes (ALP, ALT and AST) and the histological 
investigation revealed some areas of  inflammation and 
vascular degeneration which shows an evidence of  liver 
injury due to the consumption of  P.biglobosa. However 
this study provides the basis for further studies on the 
detailed toxic and pharmacological effects of  the extracts 
of  P. biglobosa stem bark and their active compounds.

RECOMMENDATION
1. There is a need for making a societal awareness on 

the pathological condition of  liver that could arise with 
unspecific consumption of  aqueous stem bark extract of  
Parkia biglobosa

2. Further research could be taken and executed using 
different extraction solvent such as Methanol or Ethanol 
to note if  there will be a significant changes in the 
diversity of  toxicity as well as the chemical constituent(s) 
of  the stem bark extract of  Parkia biglobosa

3.  A lot of  information on the adverse effects  of  the 
extract has to be establish.  

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