




































_____________________________________________________________________________________________________ 
 
*Corresponding author: E-mail: augustineairaodion@yahoo.com; 
 
 
 

Asian Journal of Immunology 
 
5(1): 40-51, 2022; Article no.AJI.85085 
 

 
 

 

 

Effect of Ethanol Extract of Xylopia aethiopica Fruit 
on Oxidative Stress Indices of Wistar Rats 

 
Emmanuel O. Ogbuagu a, Uloaku Ogbuagu b, Augustine I. Airaodion b*,  

Chika L. Uche c, Edmund O. Ezirim d, Ifeoma N. Nweke a and Prince C. Unekwee 
 

a 
Department of Pharmacology and Therapeutics, Abia State University, Uturu, Nigeria.

  

b 
Department of Biochemistry, Federal University of Technology, Owerri, Imo State, Nigeria. 

c 
Department of Haematology, Abia State University, Uturu, Nigeria. 

d 
Department of Obstetrics and Gynecology, Abia State University, Uturu, Nigeria. 

e 
Department of Pharmacology and Therapeutics, Nnamdi Azikiwe University, Awka Anambra State, 

Nigeria. 
 

Authors’ contributions  
 

This work was carried out in collaboration among all authors. All authors read and approved the final 
manuscript. 

 
Article Information 

 
Open Peer Review History: 

This journal follows the Advanced Open Peer Review policy. Identity of the Reviewers, Editor(s) and additional Reviewers, peer 
review comments, different versions of the manuscript, comments of the editors, etc are available here: 

https://www.sdiarticle5.com/review-history/85085 

 
 
 

Received 17 January 2022  
Accepted 21 March 2022 
Published 29 March 2022 

 
 

ABSTRACT 
 

Background: The use of Xylopia aethiopica fruit in folklore medicine is on the increase without 
caution of its toxicity. 
Aim: This present study tends to assess its effect on the oxidative stress biomarkers of Wistar rats. 
Methodology: The fruits of Xylopia aethiopica were air-dried and extracted by Soxhlet extractor 
using ethanol as solvent. The median lethal dose (LD50) of the extract was assessed using standard 
method. Thirty adult Wistar rats were divided into five groups of six rats each. Animals in groups 1, 
2, 3, and 4 were treated with 130, 259, 389 and 518 mg/kg body weight of X. aethiopica fruit extract 
respectively, while those in group 5 received normal animal feeds and water only. The 
administration was done once daily for 28 days via oral route. Oxidative stress indices were 
measured using standard methods. 
Results: A significant decline was observed in the antioxidant enzymes (catalase, superoxide 
dismutase, and glutathione peroxidase) activities in experimental animals compared with those in 
the control group (P<0.05). In the same vein, a significant reduction was observed in the 
concentration of reduced glutathione in experimental animals compared with those in the control 

Original Research Article 



 
 
 
 

Ogbuagu et al.; AJI, 5(1): 40-51, 2022; Article no.AJI.85085 

 
 

 
41 

 

group (P<0.05). Lipid peroxidation was however observed to increase when experimental animals 
were compared with those in the control group. The elevation in lipid peroxidation was significantly 
different when animals treated with higher doses of 259, 389 and 518 mg/kg body weight of extract 
were compared with those in the control group (P<0.05). 
Conclusion: The adverse perturbation of antioxidant indices by Xylopia aethiopica fruit is 
suggestive that it could induce oxidative stress and thus unhinged the immune system. Oxidative 
stress has also been implicated in several diseases thus the consumption of Xylopia aethiopica fruit 
as well as its use in folklore medicine should be discouraged especially in high doses due to its 
toxic nature. 
 

 
Keywords: Immune system; oxidative stress; toxic nature; Xylopia aethiopica fruit. 
 
1. INTRODUCTION 
 

Induction of oxidative stress elicits free radicals 
such as nitric oxide (NO), superoxides anions 
(O2

-
), hydroxyl radical (OH

-
), hydrogen peroxide 

(H2O2), organic hydroperoxide (ROOH) via the 
release of reactive oxygen species (ROS) [1]. 
These ROS are highly fickle atoms because they 
are known to contain a lone pair of electrons in 
their outermost shells. They are known to be 
involved in processes like aging, carcinogenesis, 
mutagenesis, and so on, which is caused by their 
in-built ability to induce cellular obstruction on 
DNA [2]. Some operations of human elicit ROS 
both endogenously and exogenously, which 
result in enervating some disease conditions. 
These diseases could arise from the adverse 
correlation in the induction of oxidative stress 
and the tendency of the living system to alleviate 
the free radicals generated sequel to the 
induction of stress [3]. Worthy of note is the point 
that the living system is built to alleviate the 
destructive nature of the radicals elicited via the 
natural antioxidants that are enzymatic in nature 
[4]. However, synthetic antioxidants also exist 
which are absorbed by the body to elevate the 
activities of the natural ones but have been 
observed to have health-related risk [2]. This has 
resulted in the renewal of vigor in the search for 
antioxidants from plant sources which will work 
well with the living system and also increase the 
ability of natural antioxidants with the purpose of 
preventing the health issues ascribed to the 
synthetics antioxidants [5]. 
 

Xylopia aethiopica has a great patronage in both 
nutrition and ethnomedicine. The plant which 
also known as African Negro pepper, is popular 
among traditional medicine practitioners and 
traditional birth attendants (TBA) who utilize the 
fruit preparations to cause the discharge of 
placental after a woman has giving birth [6]. A 

preparation of the stem bark or fruit is helpful in 
the management of bronchitis, stomach aches, 
asthma, and dysenteric conditions [6]. The seed 
extract is helpful as a vermifuge for roundworms 
[7]. Several postnatal women eat the aqueous 
preparation of the fruit for its perceived antiseptic 
properties. Some of the women have been 
reported to sometimes come to the hospitals with 
characteristics which suggest complications in 
organ [8]. Medicinal plant extracts with a 
therapeutic property has the tendency of wrong 
prescription and sometimes, overdosed. The fact 
that Xylopia aethiopica is a natural product does 
not automatically confers on it safety and might 
be risky to its consumers. Chemical ingredients 
of the plant are perceived to be useful in 
preventing and managing cancerous tumors [9]. 
Xylopia aethiopica fruit is known to have 
alkaloids, terpenoids, flavonoids, and organic oils 
[10,11]. 

 
Xylopia aethiopica is characterized with 
numerous chemical components with various 
medicinal potentials [13]. The chemical 
components of this plant have been investigated 
to include saponins, sterols, carbohydrates, 
glycosides, mucilage, acidic compounds, tannins, 
balsams, cardiac glycosides, volatile aromatic 
oils, phenols [8,14,15], alkaloids, rutin and fixed 
oils [16,17]. The plant has also be known to 
contain vitamins such as vitamin A, vitamin B, 
vitamin C, vitamin D, and vitamin E, and proteins 
as well as several minerals such as copper, 
manganese and zinc [15,17]. The impact of the 
fruit on body weight and glucose concentration of 
animals has been reported [18]. The fruit has 
also been reported to induce dyslipidemia [19], 
hepatotoxicity [20] as well as renal toxicity [21]. 
This present study focused on examining its 
impact on the oxidative stress biomarkers of 
Wistar rats. 



 
 
 
 

Ogbuagu et al.; AJI, 5(1): 40-51, 2022; Article no.AJI.85085 

 
 

 
42 

 

 

 
Fig. 1. Xylopia aethiopica Fruit [12] 

 

2. MATERIALS AND METHODS 
 

2.1 Collection and Authentication of Plant 
Materials  

 

The fruits of Xylopia aethiopica were sourced 
from a market in Aba, Abia State. They were 
identified and authenticated by Prof. Margaret 
Bassey of Botany and Ecological Studies 
Department, University of Uyo. It was assigned a 
voucher number of UU/PH/4e and deposited in 
the Herbarium of the Department of 
Pharmacognosy and Natural Medicine, 
University of Uyo, Akwa-Ibom State, Nigeria. 
 

2.2 Extraction of Plant Materials 
 

Extraction of the plant was carried out in the 
Post-graduate Laboratory of Department of 
Pharmacognosy and Natural Medicine, Faculty of 
Pharmacy, University of Uyo, Nigeria. It was 
extracted based on the outlined method in 
Ogbuagu et al. [13]. The fruits were rinsed under 
flowing tap water to eliminate contaminants and 
air-dried. The plant material was milled by 
laboratory blender. The pulverized plant material 
was macerated in 250 mL of 99.8% ethanol 
(Sigma Aldrich) contained in a flask attached to a 
Soxhlet extractor coupled with condenser and 
heating mantle (Isomantle). It was then poured 
into the sample holder (thimble) and inserted in 
the apparatus. The side arm is lagged with glass 
wool. The mixture was heated using the heating 
mantle (Isomantle) at 60 °C and as the 
temperature rises it starts to evaporate, going via 
the extractor to the condenser. The condensate 
dripped into the reservoir housing the thimble. As 
soon as the solvent gets to the siphon it emptied 
itself into the flask and the process repeats itself. 

The process goes on until it is exhaustively 
extracted. The process runs for a total of 13 
hours. As soon as it was set up, it was allowed to 
run without interruption as long as water and 
power supply were not interrupted. The 
apparatus was switched on and off and overnight 
running was not allowed, and the time for the 
complete process split over some days. The 
extract was poured into 1000 mL beaker and 
concentrated to dryness in water bath (A3672- 
Graffin Student Water Bath) at 35 °C. The total 
weight of the marc (residue) and the 
concentrated extract were noted. Several days 
was spent on the entire process. The evaporated 
extract was kept in the refrigerator until when the 
need for it arise. 

 
2.3 Determination of Median Lethal Dose 

(LD50)  

 
The median lethal dose (LD50) of the extract was 
determined using albino mice according to the 
method described by Airaodion et al. [22]. This 
method involves two phases: 

 
In Phase one, five groups containing five mice 
each weighing between 20 g and 27g were 
fasted for 18 hours. They were respectively 
treated with 1000 mg/kg, 2000 mg/kg, 3000 
mg/kg, 4000 mg/kg and 5000 mg/kg body                
weight via intraperitoneal (i.p) route and were 
monitored for visible signs of toxicity and 
mortality for 24 hours. A dosage of 1000 mg/kg 
recorded 0% mortality while 2000 mg/kg, 3000 
mg/kg 4000 mg/kg and 5000 mg/kg recorded 
100% mortality within 24 hours. Based on                  
the value of phase one, phase two was 
conducted. 



 
 
 
 

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43 

 

In Phase two, twenty-five albino mice weighing 
between 20 and 27g were grouped into 5 of 5 
mice per group and were fasted for 18 hours. 
Each group was administered 1200 mg/kg, 1400 
mg/kg 1600 mg/kg, 1800 mg/kg and 2000 mg/kg 
body weight intraperitoneally (i.p) and was 
observed for physical signs of toxicity and 
mortality within 24 hours. 1200 mg/kg recorded 
0% mortality while 1400 mg/kg, 1600 mg/kg, 
1800 mg/kg and 2000 mg/kg recorded 100% 
mortality within 24 hours. The LD50 was 
computed as geometrical means of the maximum 
dose yielding 0% mortality (a) and the minimum 
dose yielding 100% death (b).  
 

LD50 = ab  

 

2.4 Experimental Design 
 
Thirty adult Wistar rats used in this study were 
purchased from the University of Uyo, Nigeria. 
They were allowed to acclimatize for seven days 
prior to the start of the treatment. The weights 
were determined and were separated into five 
groups of six rats each. Groups A, B, C, D 
served as the experimental groups, while group 
E served as the control. Animals in group A were 
exposed to 130 mg/kg body weight (10% of LD50) 
of X. aethiopica fruit extract, those in group B 
were treated with 259 mg/kg body weight (20% 
of LD50) of X. aethiopica fruit extract, those in 
group C were exposed to 389 mg/kg body weight 
(30% of LD50) of X. aethiopica fruit extract, those 
in group D were treated with 518 mg/kg body 
weight (40% of LD50) of X. aethiopica fruit 
extract, while those in group E (control) received 
normal animals feeds and water only. The 
treatment was done once daily for 28 days via 
oral route. After 28 days treatment, the animals 
were sacrificed under ether anaesthesia in a 
desiccator after an overnight fast. Blood was 
taken from the rats through cardiac puncture.  
 

2.5 Determination of Oxidative Stress 
Indices 

 
2.5.1 Determination of superoxide dismutase 

(SOD) activity 
 
Principle: This procedure involves production of 

superoxide radical of riboflavin and its detection 
by hydroxylamine hydrochloride. The nitrite 
reacts with sulphanilic acid to yield diazonium 
compound which then reacts with naphthylamine 
to yield red azo compound whose absorbance is 
measured at 543 nm. 

This assay was done according to the method of 
Mohammad et al. [23] in which 1.4 mL aliquot of 
the reaction mixture involved 1.1 mL of 50 mM 
phosphate buffer (pH 7.4), 0.075 mL of 20 mML

-1
 

methionine, 0.4 mL of 1% (v/v) Triton X-100, 
0.075 mL of 10 mM hydroxylamine and 0.1 mL of 
50 mM EDTA. The aliquot (1.75 mL) was added 
to 0.1 mL of the sample and incubated at 30

o
C 

for 15 min. This was followed by addition of 80 
µL of 50 µM riboflavin and then the tubes were 
exposed for 9 minutes to 200 watts Philip lamp. 
After exposure time, 1 mL of Greiss reagent was 
added and the absorbance of the colour formed 
was measured at 543 nm. One unit of enzyme 
activity was measured as the amount of SOD 
capable of inhibiting 50 % of nitrate formation 
under the assay condition. 
 
2.5.2 Determination of catalase (CAT) activity 
 
Principle: The method is based on the fact that 

dichromate in acetic acid is reduced to chromic 
acetate when heated in the presence of H2O2 
with the formation of perchloric acid as unstable 
intermediate. The acetate produced is measured 
colorimetrically at 610 nm. 
 
This was assayed by the method described by 
Sinha [24]. The reaction mixture (1.5 mL) 
contained 1.0 mL of 0.01 M phosphate buffer, 0.1 
mL of the sample and 0.4 mL of 2 M H2O2. The 
reaction was stopped by the addition of 2.0 mL 
dichromate acetic acid reagent (5% potassium 
dichromate and glacial acetic acid mixed in 1:3 
ratio). Then, the absorbance was measured at 
610 nm. CAT activity was expressed as µmol of 
H2O2 consumed/min/mg protein. 
 
2.5.3 Determination of glutathione 

peroxidase (GPx) activity 
 
Principle: The activity of GPx was determined by 

measuring the decrease in GSH concentration 
after incubating the sample in the presence of 
hydrogen peroxide and sodium azide. 
 

H2O2 + 2GSH   2H2O + GSSG 

 
The activity of glutathione peroxidase was 
assayed by the method described by Rotruck et 
al. [25]. The reaction mixture contained 0.2 mL of 
0.4 M Tris- buffer, pH 7.0, 0.2 mL of EDTA, and 
0.1 mL of 10 mM sodium azide, 0.2 mL of 10 mM 
glutathione and 0.1 mL of 0.2 mM. H2O2. The 
content was incubated at 37 

o
C for 10 minutes. 

The reaction was terminated by the addition of 
0.4 ml 10% (v/v) TCA and centrifuged at 5000 



 
 
 
 

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44 

 

rpm for 5 minutes. The supernatant was assayed 
for glutathione by Ellman’s method. Exactly 3.0 
mL disodium hydrogen phosphate solution and 
1.0 mL of DNTB reagent were added to 2.0 mL 
of the supernatant. The standard was taken and 
treated in similar manner. The absorbance was 
read at 412 nm and expressed in terms of 
glutathione consumed /min/mg protein. 
 
2.5.4 Determination of lipid peroxidation  
 
Principles: Lipid peroxidation in the supernatant 

fractions was determined spectrophotometrically 
by assessing the concentration of thiobarbituric 
acid reactive substances (TBARS) as described 
by Varshney and Kale [26]. The results were 
expressed in malondialdehyde (MDA) formed 
relative to an extinction coefficient of 1.56 x 10

6
 

mol/cm.  
 
Procedure: Acetic acid 1.5 mL (20%; pH 3.5), 

1.5 mL of 0.8% thiobarbituric acid and 0.2 mL of 
8.1% sodium dodecylsulphate was added to 0.1 
mL of the sample and heated at 100 

o
C for 60 

min. After centrifugation at 1200×g for 10 min, 
the organic layer was separated and absorbance 
measured at 532nm using a spectrophotometer. 
Malondialdehyde (MDA) is an end product of lipid 
peroxidation, which reacts with thiobarbituric acid 
to form pink chromogen–thiobarbituric acid 
reactive substance. It was calculated using a 
molar extinction coefficient of 1.56×10

6 
mol/cm 

and expressed as nanomoles of MDA/tissue. The 
concentration of MDA (nmol/ml) was calculated 
by using the formula:  
 

Concentration of the test =                       

          
 

 
2.5.5 Determination of glutathione 

concentration  

 
Principles: Glutathione (reduced) was measured 

according to the method of Jollow et al. [27]. 
Reduced glutathione (GSH) forms the bulk of 
non-protein sulfhydryl groups. This method is 
based on the formation of relatively stable yellow 
colour when Ellman’s reagent is added to a 
sulfhydryl compound, 2-nitro-5-thiobenzoic acid, 
the chromophoric product resulting from the 
reaction of Ellman’s reagent with reduced 
glutathione.  
 
Procedure: Equal quantities of the sample and 

10% trichloroacetic acid were mixed and 
centrifuged at 4000 x g for 15 minutes to 
separate the proteins. To 0.5 mL of the 

supernatant, 4.5 mL of Ellman’s reagent was 
added. The mixture was vortexed and the 
absorbance read at 412 nm within 15 min.  
 

2.6 Statistical Analysis 
 

Data were subjected to analysis of variance 
using Graph Pad Prism. Results were presented 
as Mean ± Standard Error of the Mean (SEM). 
One-way analysis of variance (ANOVA) was 
used to compare the mean, followed by Tukey's 
post hoc test. Differences between means were 
considered to be significant at p<0.05. 
 

3. RESULTS 
 

3.1 Median Lethal Dose (LD50) Result 
 

The visible signs of toxicity of X. aethiopica fruit 
extract observed in this study are excitation, 
decreased motor activity, paw licking, increased 
respiratory rate, gasping and coma which could 
be followed up by death. In the first phase of the 
median lethal dose determination, no death was 
observed in the group administered 1000 mg/kg 
body weight of X. aethiopica fruit extract. 
However, all the animals died in the groups 
exposed to 2000, 3000, 4000, and 5000 mg/kg 
body weight of X. aethiopica fruit extract 
respectively (Table 1). In the same vein, in the 
second phase of medial lethal dose 
determination, no death was recorded in the 
group treated with 1200 mg/kg body weight of X. 
aethiopica fruit extract while 100% mortality was 
recorded in the groups treated with 1400, 1600, 
and 1800 mg/kg body weight of X. aethiopica 
fruit extract respectively as presented in Table 1.  
 

The median lethal dose (LD50) was computed as 
geometrical average of the maximum dose 
yielding 0% death (a) and the minimum dose 
yielding 100% death (b).  
 

LD50 =     

 

Where a = 1200 mg/kg 
 

            b = 1400 mg/kg 
 

LD50= 1296.15 mg/kg 
 

3.2 Effect of Extracts of Xylopia 
aethiopica Fruit on Oxidative Stress 
Indices of Animals after 28 Days of 
Treatment 

 

The impact of extract of Xylopia aethiopica fruit 
extract on oxidative stress parameters of animals 



 
 
 
 

Ogbuagu et al.; AJI, 5(1): 40-51, 2022; Article no.AJI.85085 

 
 

 
45 

 

after 28 days of treatment is presented in Figs. 2-
6. A significant reduction was seen in the 
activities of antioxidant enzymes (superoxide 
dismutase, catalase and glutathione peroxidase) 
in experimental animals compared with those in 
the control group (P<0.05). In the same vein, a 
noticeable reduction was seen in the 
concentration of reduced glutathione in 
experimental animals compared with those in the 

control group (P<0.05). Lipid peroxidation was 
however observed to increase when 
experimental animals were compared with those 
in the control group. The elevation in lipid 
peroxidation was significantly different when 
animals treated with higher doses of 259, 389 
and 518 mg/kg body weight of extract were 
compared with those in the control group 
(P<0.05). 

 
Table 1. The Median lethal dose (LD50) of Xylopia aethiopica fruit extract 

 

Study Phase/ 
(Animal) 

Dosage of Extract 
(mg/kg) b.w 

No of Mice 
per Group 

No. of Death 
Recorded 

% Mortality 

PHASE ONE     

I 1000 5 0 0 
II 2000 5 5 100 
III 3000 5 5 100 
IV 4000 5 5 100 
V 5000 5 5 100 

PHASE TWO     

I 1200 5 0 0 
II 1400 5 5 100 
III 1600 5 5 100 
IV 1800 5 5 100 
V 2000 5 5 100 

LD50= 1296.15 mg/kg 

 

13
0 

m
g/k

g X
. a

et
hio

pic
a 

25
9 

m
g/k

g X
. a

et
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pic
a

38
9 

m
g/k

g X
. a

et
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a

51
8 

m
g/k

g X
. a

et
hio

pic
a

C
ontr

ol
0

1

2

3

Treatment Groups

A
c
ti

v
it

y
 o

f 
S

O
D

 (
U

/m
L

)

 

 
Fig. 2. Effect of X. aethiopica fruit extract on the Activity of Superoxide Dismutase (SOD) in 

Animals after 28 days of Treatment 
Each bar represent mean ± SD of treatment groups with n = 6 



 
 
 
 

Ogbuagu et al.; AJI, 5(1): 40-51, 2022; Article no.AJI.85085 

 
 

 
46 

 

13
0 

m
g/k

g X
. a

et
hio

pic
a 

25
9 

m
g/k

g X
. a

et
hio

pic
a

38
9 

m
g/k

g X
. a

et
hio

pic
a

51
8 

m
g/k

g X
. a

et
hio

pic
a

C
ontr

ol
0

1

2

3

4

5

Treatment Groups

A
c
ti

v
it

y
 o

f 
C

a
ta

la
s
e
 (

U
/m

L
)

 

 
Fig. 3. Effect of X. aethiopica fruit extract on the Activity of Catalase in Animals after 28 days of 

Treatment 
Each bar represent mean ± SD of treatment groups with n = 6 

13
0 

m
g/k

g X
. a

et
hio

pic
a 

25
9 

m
g/k

g X
. a

et
hio

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a

38
9 

m
g/k

g X
. a

et
hio

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a

51
8 

m
g/k

g X
. a

et
hio

pic
a

C
ontr

ol
0

1

2

3

4

Treatment Groups

A
c
ti

v
it

y
 o

f 
G

P
x
 (

u
/m

L
)

 

 
Fig. 4. Effect of X. aethiopica fruit extract on the Activity of Glutathione Peroxidase (GPx) in 

Animals after 28 days of Treatment 
Each bar represent mean ± SD of treatment groups with n = 6 

 



 
 
 
 

Ogbuagu et al.; AJI, 5(1): 40-51, 2022; Article no.AJI.85085 

 
 

 
47 

 

13
0 

m
g/k

g X
. a

et
hio

pic
a 

25
9 

m
g/k

g X
. a

et
hio

pic
a

38
9 

m
g/k

g X
. a

et
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pic
a

51
8 

m
g/k

g X
. a

et
hio

pic
a

C
ontr

ol
0

1

2

3

4

5

Treatment Groups

C
o

n
c
. 

o
f 

G
S

H
 (

m
M

)

 

 
Fig. 5. Effect of X. aethiopica fruit extract on the Concentration of Reduced Glutathione (GSH) 

in Animals after 28 days of Treatment 
Each bar represent mean ± SD of treatment groups with n = 6 

 

13
0 

m
g/k

g X
. a

et
hio

pic
a 

25
9 

m
g/k

g X
. a

et
hio

pic
a

38
9 

m
g/k

g X
. a

et
hio

pic
a

51
8 

m
g/k

g X
. a

et
hio

pic
a

C
ontr

ol
0

2

4

6

8

Treatment Groups

C
o

n
c
. 

o
f 

L
P

O
 (

u
M

)

 

 
Fig. 6. Effect of X. aethiopica fruit extract on Lipid Peroxidation of Animals after 28 days of 

Treatment 
Each bar represent mean ± SD of treatment groups with n = 6 

 



 
 
 
 

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48 

 

4. DISCUSSION 
 
The acute toxicity study of X. aethiopica fruit 
extracts led to 100% death at a dose of 1400 
mg/kg bodyweight and above. This reveals that 
this fruit could be greatly toxic. The visible 
symptoms of toxicity seen in the animals were 
excitation, paw licking, increased respiratory rate, 
decreased motor activity, gasping and coma and 
death.  
 
In this study, a noticeable decrease was 
recorded in superoxide dismutase (SOD) activity 
in animals exposed to Xylopia aethiopica fruit 
extract at all tested doses when compared with 
control animals at P<0.05 (Fig. 2). This 
suggested that Xylopia aethiopica fruit extract 
might be toxic and could possibly play a role in 
the production of free radicals. This agreed with 
the results of Nnodim et al. [28] who observed 
that Xylopia aethiopica fruit induced oxidative 
stress. 

 
Observation from this study showed that 
treatment with ethanol extract of Xylopia 
aethiopica fruit was observed to greatly reduce 
the activity of catalase in animals treated with 
259, 389 and 518 mg/kg of Xylopia aethiopica 
fruit extract when compared to animals in the 
control group at P<0.05 (Fig. 3). This might be an 
indication that Xylopia aethiopica fruit extract at 
these doses might be toxic and possibly lead to 
induction of oxidative stress. This agreed with the 
findings of Nnodim et al. [28], who reported that 
Xylopia aethiopica fruit generated free radicals in 
treated animals. 

 
Treatment with extract of Xylopia aethiopica fruit 
significantly down-regulated glutathione 
peroxidase (GPx) activity at all tested doses (Fig 
4). The biological role of glutathione peroxidase 
is to change hydroperoxides of lipid to their 
respective alcohols and release hydrogen 
peroxide to form water [29,30]. This is in 
consonance with the results of Nnodim et al. [28], 
who reported that Xylopia aethiopica fruit 
significantly reduced the activity of glutathione 
peroxidase of treated animals. Glutathione 
peroxidase is one of the enzymatic antioxidant 
with the capacity to defend the system and 
combat oxidative stress [31]. The reduction in the 
activity of glutathione peroxidase observed in this 
study might be an indication that Xylopia 
aethiopica fruit has the propensity to induce 
oxidative stress. 
 

In this study, administration of ethanol extract of 
Xylopia aethiopica fruit resulted in a decline in 
the concentration of reduced glutathione at all 
doses when compared with control group (Fig. 
5). The noticeable decrease observed in the 
reduced glutathione level in this study might be 
an indication that Xylopia aethiopica fruit 
enhances the conjugation of GSH with 
acetaldehyde [32]. This may have resulted from 
the direct reactive oxygen species (ROS) 
producing potential of Xylopia aethiopica fruit 
and/or a decrease in GSH synthesis. 
 

In this study, administration of Xylopia aethiopica 
fruit extract was seen to have noticeably led to a 
surge in the malondialdehyde (MDA) levels 
indicating increased peroxidation and catabolism 
of the antioxidant defense mechanisms. 
Malondialdehyde is a product of lipid 
peroxidation of polyunsaturated fatty acids 
[33,34]. Furthermore, visible destruction of 
tissues in lipid peroxidation caused by free 
radicals could cause membrane injury followed 
by decrease in the membrane fluid content. This 
is in line with the study of Nnodim et al. [28], who 
observed that Xylopia aethiopica fruit significantly 
elevated lipid peroxidation (Fig. 6). 
 

The depletion in antioxidant enzymes activities 
seen in this study agrees with the work of 
Somnez et al. [35]. The biochemical mechanism 
by which Xylopia aethiopica caused reduction in 
enzymatic antioxidants is currently unclear. It 
might be postulated that the consumption of this 
plant extract could cause oxidative stress and 
hence generate free radicals which could result 
in membrane destruction through lipid 
peroxidation and protein oxidation [36-38]. Thus, 
the reduction in the activities of superoxide 
dismutase (SOD), catalase (CAT), as well as 
glutathione peroxidase (GPx), elevated lipid 
peroxidation. This became visible by the 
increased level of MDA observed in this present 
investigation. 
 

5. CONCLUSION 
 

The adverse perturbation of antioxidant indices 
by Xylopia aethiopica fruit is suggestive that it 
could induce oxidative stress and thus negatively 
impact the immune system. Oxidative stress has 
also been implicated in different diseases, thus 
the consumption of Xylopia aethiopica fruit as 
well as its use in folklore medicine should be 
discouraged especially in high doses due to its 
toxic nature. 
 



 
 
 
 

Ogbuagu et al.; AJI, 5(1): 40-51, 2022; Article no.AJI.85085 

 
 

 
49 

 

CONSENT 
 
It is not applicable. 

 
ETHICAL APPROVAL 
 
As per international standard or university 
standard written ethical approval has been 
collected and preserved by the author(s). 

 
ACKNOWLEDGEMENT 
 
Authors are grateful to Mr. Nsikan Malachy for 
his assistance in the practical aspect of this work. 
 

COMPETING INTERESTS 
 

Authors have declared that no competing 
interests exist. 
 

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