ORIGINAL�ARTICLE

ABSTRACT
Objective: To compare the hepatoprotective effects of aqueous extract of Cassia fistula leaves and Silymarin on 
Acetaminophen induced hepatotoxicity.
Study Design:  An experimental study in mice.
Place and Duration of Study: The study was conducted in the department of Pharmacology and Therapeutics, 
Multidisciplinary research laboratory of Islamic International Medical College, Riphah International University. 
The duration of study was from 16 Mar, 2016 to 16 Mar, 2017.
Materials and Methods: In present study simple random sampling was done by Balloting method and sixty 
adult Balb/c Albino mice were divided in four groups having 15 mice each. Mice in control group A were given 
normal diet and fresh water. Mice in experimental group B (positive control) were given Acetaminophenat dose 
of 100mg /kg /day orally for six weeks. Mice in experimental group C were treated with 
Acetaminophen(100mg/Kg) with concurrent  400mg/Kg body weight of Cassia fistula (Aqueous extract) orally 
for six weeks once daily while in experimental group D mice received 750 mg/kg body weight of Silymarin orally 
in form of aqueous suspension in 2% Cremophor1E for 6 weeks once daily concurrently treated with 
Acetaminophen(100mg/Kg).The cardiac puncture was done after six weeks  and 2.5cc blood was collected 
from each mice for assessment of biochemical markers including Serum Alanine Aminotransferase (ALT) and 
Serum  Aspartate Aminotransferase (AST).
Results:  The mean value of serum ALT level was 130.4.30 ±42.90 in Group B as compared to control Group 
having mean value of  ALT  37.30 ± 8.32. The mean value of ALT was 62.22± 19.27 and 53.90± 25.06 in Group C 
and in Group D respectively. In Group B mean value of serum level of  AST was  134.3± 15.56 as compared to 
control Group A having mean value of AST31.40± 10.05.The mean value of AST in Group C and Group D were 
66.40±20.50 and 44.67± 20.37 respectively.
Conclusion: The Silymarin is more beneficial than aqueous extract of Cassia fistula leaves  in ameliorating 
hepatotoxic effects of Acetaminophen. 

Key Words: Acetaminophen, Cassia fistula leaves, Hepatoprotective, Silymarin.

considered toxic to liver. The incidence of DILI is 1 in 
4 

10,000 to 1 in 100,000.   In humans the most 
common cause of drug induced liver failure is 
a c e t a m i n o p h e n  ( Pa ra c e t a m o l ,  N - a c e t y l - p -

5
aminophenol,APAP) toxicity.  It is safe at therapeutic 
doses but at larger doses it can cause serious liver 

6,7 
injury. The cytochrome P-450 oxidation of 
acetaminophen overdose result in formation of 
reactive metabolite N-acetyl-p-benzoquinone imine 

8 
(NAPQI) which initiates toxicity. It depletes 
glutathione and binds to cellular proteins, especially 

7,9,11
in mitochondria and cause hepatotoxicity.  
Previous studies have also identified NAPQI as 
important factor in acetaminophen induced 

8 
hepatotoxicity. Previous studies have identified 
s o m e  h e p a t o p r o t e c t i v e  a g e n t s  a g a i n s t  
acetaminophen induced hepatotoxicity.  Silymarin 

Introduction
Liver is a vital organ having role in metabolism of 
carbohydrates, proteins and lipids, bile production, 
maintenance of homeostasis, detoxification and 

1,2
vitamin storage.  Drug induced liver injury (DILI)  is 
the leading cause of acute liver failure in most 

3 
countries. There are more than 1000 drugs 

Comparing Hepatoprotective Effects of Aqueous Extract of Cassia 
Fistula (Amaltas) Leaves and Silymarin 
Rukhsana Munawar, Adnan Jehangir, Akbar Waheed

Correspondence:
Dr. Rukhsana Munawar
Department of Pharmacology
Islamic International Medical College
Riphah International University, Islamabad
E-mail: dr.rukhsana33@gmail.com

Department of Pharmacology
Islamic International Medical College
Riphah International University, Islamabad

Funding Source: NIL; Conflict of Interest: NIL
Received: Dec 27, 2017; Revised: Jul 09, 2018
Accepted: Jul 16, 2018

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137



leaves were boiled in distilled water for 30 minutes. 
These were kept for 3 days with intermittent shaking. 
These were filtered by Whatman No1 filter paper. 
The aqueous extract was obtained by concentrated 

17,19
the filtrate by rotary flash evaporator.
The mice were divided into four groups randomly, 
each group consisting of 15 mice. Mice in Control 
Group A were given normal diet and fresh water 
o r a l l y.  E x p e r i m e n t a l  G r o u p  B  w a s  g i v e n  
Acetaminophen 100mg/Kg orally  in diet  for 6 weeks 

20 
once daily. Experimental Group C was treated with 
acetaminophen 100mg/Kg  with concurrent  400 
mg/kg body weight of Cassia fistula(Aqueous 

21 
Extract) orally for 6 weeks once daily. Experimental 
Group D was  treated with acetaminophen 
100mg/Kg orally with concurrent 750 mg/kg BW of 
Silymarin in form of aqueous suspension in 2% 

22
Cremophor1E for 6 weeks once daily.  After six 
weeks cardiac puncture was done and blood samples 
were taken for evaluation of Serum Alanine 
Aminotransferase (ALT) and Serum Aspartate 
Aminotransferase (AST) levels.
The data was analyzed by using Statistical package 
for social sciences (SPSS) version 22. The biochemical 
parameters ALT and AST were calculated by using 
Mean ± Standard deviation (SD). The mean 
difference between control and other groups was 
calculated by applying ANOVA. The Post Hoc Tuckey 
test was applied for comparison of mean difference 
between groups. p-value 0f <0.05 was considered 
statistically significant.

Results
The serum ALT and AST levels were significantly 
raised (p<0.00) in Acetaminophen treated Group B 
as compared to control Group A. The serum ALT and 
AST were reduced in  Cassia fistula treated Group C 
and Silymarin treated Group D and in comparison, 
the results showed significant reduction in serum 

(SLM) is one of them having antioxidant, anti-
i n f l a m m a t o r y,  a n t i f i b r o t i c ,  a n t i v i r a l  a n d  

1,12,14 
immunomodulatory actions. The Silymarin has 
hepatoprotective and antioxidant role by  inhibiting  

1,15
free radicals.  
 Herbal medicines represent one of the most 
important fields of traditional medicine throughout 

 
the world. Cassia fistula (Family Leguminosae, Sub-
family Caesalpinea) is a very common plant known 
for its medical properties. It is commonly called 
Amaltas. The antioxidant property is possessed by 

16 
polyphenol and flavonoids.
 Previous studies showed work done on methanolic 
and ethanolic extracts of Cassia fistula leaves. In 
present study aqueous extract was used to explore 
hepatoprotective effect of Cassia fistula leaves and 
its hepatoprotective effect was compared with 
Silymarin on Acetaminophen induced hepatotoxicity 
in mice and this comparison was not found in 
previous studies.
The aim of present study was to compare 
hepatoprotective effects of Cassia fistula leaves and 
Silymarin on acetaminophen induced hepatotoxicity.

Materials and Methods
An experimental study was carried out in the 
department of Pharmacology and Therapeutics, 
Multidisciplinary research laboratory of Islamic 
International Medical College in collaboration with 
National Institute of Health Sciences (NIH), 
Islamabad from Mar 2015 to Mar 2016. Approval of 
synopsis was authorized by the Ethical Review 
Committee of Islamic International Medical College. 
Mice weighing 30-50grams with normal ALT and AST 
levels were included in the study and mice weighing 
less than 30grams and more than 50grams with 
abnormal liver function tests were excluded.  Sixty 
Balb/c healthy albino mice were selected randomly 
by balloting method.   A perfect living condition 
indistinguishable to their class with adjusted dietary 
supplement was given under temperature of 24±2ºC 
with a 12 hour light and dark cycle. 
Cassia fistula leaves were collected from Race course 
park, Rawalpindi. The leaves were air dried and then 
authenticated by herbarium department of National 
Agricultural Centre, Islamabad. The external dirt of 
freshly collected leaves was removed by washing 
with tap water. The leaves were shade dried and 
crushed coarsely by hand. About 400 gm of these 

Table I: Comparison of Serum ALT Level between the 
Groups (n=60)

P value < 0.00  P value <0.05 =significant

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In present study significant improvement (p < 0.05) 
in serum levels of ALT and AST in Group D is observed 
which was given Acetaminophen followed by 
Silymarin. This was also revealed by study of Amir 
Mohammad Kazemifar et al., who demonstrated the 
hepatoprotective property of Silymarin in 
Acetaminophen induced hepatotoxicity in male 

25
Sprague-Dawley rats.
In present study a significant reduction (p < 0.05) was 
observed in raised serum levels of ALT and AST in 
Group C which was given Acetaminophen followed 
by Cassia fistula as compared to Group B which was 
given Acetaminophen. The hepatoprotective effect 
of ethanolic extract of Cassia fistula leaves was 
observed in study of  Pradeep et al. at dose of 
500mg/kg/day for 7 days against subacute Carbon 
Tetrachloride (CCl4) induced hepatic injury in rats. He 

17 
found significant improvement in levels of ALT, AST.
Das et al. also proved the hepatoprotective effect of 
aqueous extract of fruit pulp of Cassia fistula against 

18
liver damage induced by CCl4 in Albino rats.  Our 
findings are in accordance with study of  Jehangir et 
al. who obser ved significant reduction in 
biochemical parameters including ALT, AST, ALP and 
total bilirubin  by ethanolic extract of Cassia fistula 
leaves against hepatotoxicity induced by Isoniazid 

27 
and Rifampicin. The better hepatoprotective effect 
of aqueous extract of Cassia fistula leaves could be 
proven with increasing duration of study. Further 
studies are required to see hepatoprotective effect 
of Cassia fistula leaves with higher doses and by use 
of different routes of administration. The molecular 
mechanism responsible for reducing biochemical 
and histopathological parameters can be explored by 
further studies.

Conclusion
In present study significant hepatoprotective effect 

was shown by aqueous extract of Cassia fistula 

leaves probably due to presence of flavonoids. 

Silymarin demonstrated better hepatoprotective 

effect than Cassia fistula against Acetaminophen 

induced hepatotoxicity.

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Table II: Post hoc Tukey's test for mul�ple comparison 
of ALT between the Groups

Table III: Post hoc Tukey's Test for Mul�ple Comparison 
of AST between the Groups

Hepatoprotective Effects of Cassia Fistula Leaves & Silymarin JIIMC 2018 Vol. 13, No.3

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Hepatoprotective Effects of Cassia Fistula Leaves & Silymarin JIIMC 2018 Vol. 13, No.3

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