ORIGINAL�ARTICLE

ABSTRACT
Objective:  To study the protective effect of olive oil against histological changes induced by arsenic in liver of 
albino rats.
Study Design: Randomized control trail.

st th
Place and Duration of Study: The research was carried out from 1  to 30  November 2017 at National institute 
of Health, Islamabad.
Materials and Methods: Forty five male adult albino rats were placed in three cages having 15 rats each. 
Distilled water was given to rats of control group I for 30 days. The rats of group II were given 40mg per kg per 
day of Sodium Arsenite dissolved in drinking water for 30 days. Rats in group III, in addition to sodium Arsenite 
received olive oil, 0.2ml per day for 30 days along with Sodium Arsenite. Dissection was done after 30 days and 
liver was dissected out for histological changes.
Results: The use of olive oil improved the gross and microscopic changes induced by Arsenic in liver lobes(right 
lateral and left lateral) of Albino rats of group III as compared to group II rats, which received only arsenic. 
Among microscopic parameters, sinusoidal dilation, pyknosis and necrosis was markedly reduced by use of 
olive oil in group III rats whereas hemorrhage was absent in group III.
Conclusion: Olive oil protects histological changes caused by arsenic in liver of Albino rats which include 
sinusoidal dilation, congestion, pyknosis, necrosis and haemorrhage.

Key Words: EVOO (extra virgin olive oil), Sodium arsenite, Oleuropein.

species (ROS)” and “reactive nitrogen species (RNS)”  
leading to necrosis, oxidative damage to proteins, 

1,3,4 
lipids and DNA in cells. Several acute and chronic 
hepatic effects have been associated with arsenic 

5
poisoning.  Various antioxidants are available which 
can reduce the effect of arsenic on liver and various 
organs, olive oil is one of them which can be used to 
avoid  disastrous effects of arsenic on liver. Olive oil 
contains about 70% Oleic acid and phenolic 
compounds that provide health benefits. Olive  oil 
itself has a greater antioxidant capacity than most 

6
other seed oils.
Arsenic affects most of the organs involved in 

7
absorption, accumulation and excretion,  Long-term 
exposure to inorganic arsenic can cause dysfunction 

8,9 10
of endocrine system  nervous system,  and 

11
reproductive system,  and may  also  cause loss of 

4
body weight.  Exposure to arsenic also causes, liver 

12,13
fibrosis, metabolic disorder such as diabetes,  

9
chronic lung disease, gangrene of toes,  cancer of 

14
internal sites and skin.  It has been found to have 
toxic effects on gonadal tissues of laboratory animals 

11
as well.
Various antioxidants have been used to ameliorate 

Introduction
Liver, the largest organ in the body, usually weighs 
about 1.5 kg. It is an organ of metabolism and 
production of energy; its other main functions 
include: storage of iron, trace elements, vitamins and 
bile production. The weight of human liver is 2 to 3%, 

1
whereas rat liver is 5% of total body weight.  Various 
metals have acute and chronic effects on liver; 
Arsenic is one of them which also produce toxic 
effects on liver. Arsenic, a “protoplasmic poison” 
interferes with mitosis, cell respiration, enzymes due 

2
to its effect on sulfhydryl group of cells.  It can also 
exert its toxic effects by generating “reactive oxygen 

The Protective Effect of Olive Oil on Arsenic Induced Histological
Changes in the Liver of Albino Rats  

1 2
Hira Waqas Cheema , Shabana Ali  

Correspondence:
Dr. Hira Waqas Cheema
Senior Lecturer 
Department of Anatomy
Islamic International Medical College
Riphah International University, Islamabad
E-mail: hira.cheema@hotmail.com

1,2
Department of Anatomy

Islamic International Medical College
Riphah International University, Islamabad

Funding Source: NIL; Conflict of Interest: NIL
Received: April 30, 2018; Revised: November 19, 2018
Accepted: December 16, 2018

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toxic effects of arsenic on various organs. Vitamin E, 
Ca and Olive oil is used to improve Arsenic induced 

15,6
histological  changes in ovary.  Antioxidants like L 
ascorbic acid, biochanin A, menthe piperita and aloe 
Vera has been used to improve toxic effects of 

16,19 
arsenic on liver of rats. Oleuropein, one of the 
component of olive oil is found to be effective 
antioxidant in literature, therefore olive oil may be 
used in our study to ameliorate arsenic induced 

20
hepatotoxic effects.  The present study was 
designed to study the protective effect of olive oil 
against histological changes induced by arsenic in 
liver of albino rats.

Materials and Methods
The experiment was carried on the basis of 
randomized control trial under supervision of animal 

st th
house at NIH Islamabad from 1  to 30  November 
2017. Forty five male albino rats, weighing 250 to 
300gm were kept in three cages with a number of 15 
rats per cage. The simple random sampling 
technique was used. The research was approved by 
Ethical Review Committee. A controlled standard 
living environment suitable to their class with 
adjusted diet was given. A well ventilated room with 
cycles of 12 hours light and 12 hours dark were 

0
maintained under 20 to 26 C. The rats were adult of 
age 2 to 4 months and those with any known 
pathology and female rats were excluded. Animals 
were grouped accordingly as mentioned below.

sections. The slides were examined in detail under 
X10 and X40 power of light microscope. The 
microscopic qualitative parameters were observed 
which include Sinusoidal dilation, Congestion, 
Pyknosis, Necrosis and haemorrhage

Results
In control group I, experimental animals showed 
normal sinusoids while in group II and group III 
sinusoidal dilation was present in 100% of 
experimental animals. The use of olive oil in group III 
has significantly reduced the severity and amount of 
sinusoidal dilation caused by arsenic Table 1 and 
Figure 1). 

After accomplishment of 4 weeks duration of 
experiment, rats were anesthetized with chloroform 
and dissected. After fixation and embedding, eosin 
and hematoxylin stains were used for histological 

Group I

Group II

Group III (mild sinusoidal dila�on)
Fig 1: Group Wise Distribu�on of Sinusoidal Dila�on of 
Hepa�c Lobule Showing Normal Sinusoid in Group I L7B1,
Severe Sinusoidal Dila�on in Group II L4D and Mild 
Sinusoidal Dila�on in Group III L6 A.(H and E,X 40) 
(Indicated by Arrows in Group 1 and Group II of Figure 1)

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The control group showed no congestion in sinusoids 
and central vein. 100 % of experimental animals in 
group II showed congestion mainly in central vein 
and also in sinusoids,whereas 46.7% of rats in group 
III showed congestion but 53.3% showed no 
congestion.In this way olive oil has  decreased the 
number of rats in group III showing congestion. 
(figure 2)

There was normal size of nucleus of hepatocytes in 
the control group I, pyknosis was present in 100 % of 
experimental animals of group II,Olive oil in group III 
has significantly reduced the number of rats showing 
pyknosis to 60%.(Table 4) (figure 3). 

Table I ꞉Group Wise Distribu�on of Sinusoidal Dila�on in 
Hepa�c Lobule Among Control and Experiental Groups of 
Albino Rats

Group I

Group II Conges�on in Central Vein and Sinusoids

Group III

Fig 2: Group Wise Distribu�on of Conges�on of Sinusoids 
and Central Vein Showing Normal Sinusoids and Central 
Vein in Group I L7D, Conges�on in Central Vein and 
Sinusoid in Group II L4B, Whereas no Conges�on in 
Group III L3C. (H and E, X 40). (Indicated by Arrow Heads 
in Group II Figure 2)

Table II: Group Wise Distribu�on of Conges�on of 
Sinusoids and Central Vein Among Control and 
Experimental Groups

Group I

Group II

Group III

Fig 3: Group Wise Distribu�on of Pyknosis of 
Hepatocytes Among Control and Experimental Groups 
Shows Normal Nuclear Size in Group IL7A, Presence of 
Pyknosis in GroupII L4D and Absence of Pyknosis in 
Group III L3 B(H and E,X 40) .(Indicated by Circles in 
Figure 3)

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The normal hepatic parenchyma was observed in 
group I while 100% of experimental rats in group II 
showed necrosis in hepatic parenchyma, whereas 
use of olive oil along with arsenic in group III has 
significantly reduced the necrosis to 53.3% of rats in 
group III

In group I, hepatic parenchyma appeared to be 
normal, 46% of rats in group II showed hemorrhage. 
In group III, olive oil has significantly improved 
hemorrhage as it was absent in all rats. (Table 
IV)(Figure 6)
In group I, hepatic parenchyma appeared to be 
normal, 46% of rats in group II showed hemorrhage 
group III, olive oil has significantly improved 
hemorrhage as it was absent in all rats.(Table 

Table III: Group Wise Distribu�on of Pyknosis of 
Hepatocytes among Control and Experimental Groups 
of Albino Rats

Group I

Group II (periportal necrosis)

Group III

Fig 4: Group Wise Distribu�on of Necrosis of Hepa�c 
Parenchymma among Rat Groups Shows Normal Hepa�c 
Parenchyma in Group IL7A2, Presence of Coagula�ve 
Necrosis in Periportal Areas in Group II L5B, and Absence 
of Necrosis in Group III L3A(H and E,X 40). (Indicated 
by Arrow Head in Group II Figure 4)

Group I

Group II

Group III

Fig 5: Bar Chart Showing Distribu�on of Necrosis of 
Hepa�c Parenchyma among Rat Groups

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IV)(figure6)
Figure 6�Group wise distribution of haemorrhage in  
hepatic  parenchyma among groups 
shows normal parenchyma  in group I L7 
C,haemorrhage indicated by arrow in group II L4D 
whereas no haemorrhage in group III L6A.(H and E, X 
40). 

reduced congestion to only 46.7% of rats. In a study 
by Azab, when olive leave extract was given along 
with carbendazim, then congestion in central vein 
and sinusoids was remarkably decreased as 
compared to the group which was given only 

25
carbendazim.
There was normal size of nucleus of hepatocytes in 
the control group I, pyknosis was present in 100 % of 
experimental animals of group II, Olive oil in group III 
has significantly reduced the number of rats showing 
pyknosis to 60%. In a study conducted by Somia 
Bashir of India, arsenic was given to three groups in 
three different doses for acute period of 24 hours 
and pyknosis was found only in group which was 
given higher dose. This study support that Pyknosis is 

26
early sign of necrosis.
Necrosis occurs as a result of inflammation of liver 
caused by arsenic and it occurs after sinusoidal 
dilation and congestion in which degeneration of 
hepatocytes takes place due to ischemia. In the 
recent study, there is absence of necrosis in 
experimental animals of group I while 100% of 
experimental rats in group II showed necrosis 
whereas 53.3% of rats in group III showed necrosis. In 
this way olive oil has significantly decreased necrosis. 
In group II, there was periportal necrosis which is 
zone I as well as zone III which is pericentral, least 
oxygenated. In group III treated by both arsenic and 

27
olive oil, there is only pericentral necrosis.  In one of 
the study carried out by Sujata Das, sodium arsenite 
was also given in dose of 40mg per kg in drinking 
water to mice for 30 days and he found periportal 

28
necrosis similar to recent study.  The use of olive oil 
in group III has improved necrosis in our study. Metin 
Ogun, used oleuropein in dose of 30mg per kg along 
with 5mg per kg of sodium arsenite for only 15 days 
and he found that group which was given only 
arsenic ,6 mice showed necrosis but the  oleuropein  
group showed necrosis in only 1 mouse. 
The haemorrhage occurs late in process of 
inflammation due to damage of endothelial lining of 
sinusoids leading to extravasation of blood into 
parenchyma. The livers of group I showed normal 
parenchyma but in group II, 46% of rats only show 
hemorrhage as arsenic was given to them for 30 
days. In group III, hepatic parenchyma was normal 

29
with no hemorrhage.  Daqian Yang in his study gave 
As O  (arsenic trioxide) intraperitoneally in a dose of 2 3

Table IV: Group Wise Distribu�on of Haemorrhage in 
Hepa�c parenchyma among  among Control and 
Experimental Groups of Albino Rats

Discussion
The sinusoidal dilation was observed in groups I,II 
and III. In control group I, experimental animals 
showed normal sinusoids while in group II and group 
III sinusoidal dilation was present in 100% of 
experimental animals. The use of olive oil in group III 
has significantly reduced the sinusoidal dilation as 
compared to dilation in group III caused by arsenic. In 
one study carried out by Kharroub et al, sodium 
arsenite was given at 1mg and 10mg for 45 and 90 
days respectively which showed that sinusoidal 
dilation is dose and duration dependent. The dilation 
was greater at 10mg of arsenic for 90 days as 

21
compared to 1mg at 45 days.  In another study 
conducted by Sohini Singh,4 to 10mg of arsenic was 
given for 30 days to rats which also caused sinusoidal 

16
dilation of liver.  In group III, there was slight 
sinusoidal dilation which is due to hepatoprotective 
effect of olive oil given simultaneously with arsenic 
to this group. Farag in his study showed natural 
antioxidant effect to prevent sinusoidal dilation and 

22
congestion caused by oxidative damage to liver.  
Congestion of sinusoids and central vein is caused 
due to inflammation of liver. All rats of group II 
showed congestion mainly in central vein and also in 
sinusoids due to effect of arsenic.46.7% of rats of 
group III showed congestion but 53.3% showed no 

23
congestion.  In the study conducted by Oyagbemi et 
al, arsenic was given to three different groups at 
different doses for 4 weeks, they showed congestion 

24
in hepatic vessels both in central vein and sinusoids.  
The use of olive oil in group III has significantly 

*P≤0.05

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3mg per kg for 2 weeks to rats and he found 
hemorrhage in liver histological examination. When 
olive oil was given to group III along with arsenic, due 
to its antioxidant effect it prevents the liver of rat to 
develop hemorrhage. In support of ameliorating 
effect of olive oil to improve hemorrhage of liver of 
our study, olive leave extract was used to improve 
hemorrhage in liver caused by carbendazim in a 

25
study done by Azab of Libya.

Conclusion
Olive oil protects histological changes caused by 
arsenic in liver of albina rats, which include 
sinusoidal dilation, congestion, pyknosis, necrosis 
and haemorrhage.

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