ORIGINAL�ARTICLE

ABSTRACT
Objective: To evaluate the effects of Aqueous and Methanolic extracts of Cichorium Intybus seeds on 
Gentamicin induced nephrotoxicity in rats.
Study Design: Experimental Study
Place and Duration of Study:  Study was conducted from 15 January to 31 March 2017 at National Institute of 
Health Sciences (NIH) in collaboration with Riphah Institute of Pharmaceutical Sciences (RIPS).
Materials and Methods: Forty healthy Sprague Dawly rats weighing 300-350gms were randomly divided in four 
groups with 10 rats each. Group A was the control group that received no medications. Group B was given 
Gentamicin 80mg/ kg body weight / day for 10 days intraperitoneally (IP). Group C was given Gentamicin 80mg/ 
kg body weight / day IP and Aqueous Extract of Cichorium Intybus seeds 500mg/kg body weight/ day through 
gavage tube for 10 days. Group D was given Gentamicin 80mg/ kg body weight / day IP and Methanolic Extract 
of Cichorium Intybus seeds 500mg/kg body weight / day through gavage tube for 10 days. Serum urea and 
creatinine were measured at day zero, 6, and 11. The values were compared within and between the groups.
Results: Elevated levels of serum urea and creatinine were found in group B indicating renal damage. Whereas 
administration of Aqueous and Methanolic extracts of Cichorium Intybus seeds reduced the elevated levels of 
serum urea and creatinine in group C & D. However more reduction was produced in group C.
Conclusion: Concomitant administration of Aqueous and Methanolic extracts of Cichorium Intybus seeds 
exerted nephroprotective effect on Gentamicin induced nephrotoxicity in rats. Aqueous extract produced 
greater protection as compared to Methanolic extract.

Key Words: Cichorium Intybus Seeds, Gentamicin, Nephroprotective effect, Nephrotoxicity.

therapeutically used drugs may result in acute or 
2 

chronic renal failure. Across the globe approximately 
19-25% cases of nephrotoxicity in critically ill 

1  
p at i e nt s  re s u l t s  f ro m  n e p h ro tox i c  d r u g s .
Nephrotoxicity is indicated by the raised levels of 

3 
serum urea and creatinine. Drug groups which cause 
nephrotoxicity include immunosuppressant, 
anticancer drugs, radio contrast media and 

.4
antibiotics
Aminoglycosides (AGs) is among one the widely 
prescribed antibiotic group throughout the world 

4  
being cost effective. Aminoglycosides are 
bactericidal antibiotics which irreversibly inhibit 
protein synthesis by binding to specific 30S-subunit 
ribosomal protein through interference of initiation 
of complex of peptide, breaking of polysome into 

5
monosomes and misreading of mRNA.  The major 
adverse effects caused by aminoglycosides are 
ototoxicity and nephrotoxicity, which limits their use. 
Aminoglycosides are excreted unchanged through 

7 
glomerular filtration without metabolism. During 
the process of concentration and reabsorption 

Introduction
Kidneys play a vital role in the body performing 
important functions for example excretion of 
metabolic waste, drugs, homeostasis maintenance 

1
and extracellular environment regulation.  Direct or 
indirect exposure of drugs and different chemicals to 
k i d n e y s  r e s u l t s  i n  n e p h r o t o x i c i t y.  M a n y  

Effects of Aqueous and Methanolic Extracts of Cichorium Intybus
Seeds on Gentamicin Induced Nephrotoxicity in Rats

1  2  3  4  5 6
Muhammad Tahir , Noman Sadiq , Sameer Ahmed , Amanat Ali , Noor Nasir Rajpoot , Uzma Riaz

Correspondence:
Dr. Muhammad Tahir
Assistant Professor
Department of Pharmacology
Women Medical and Dental College, Abbottabad
E-mail: m.tahir9044@gmail.com

1
Department of Pharmacology

Women Medical and Dental College, Abbottabad
2
Departmant of Physiology

CMH Kharian Medical College, Kharian
3,4

Departmant of Pharmacology
HBS Medical and Dental College, Islamabad
5
Department of Medicine

Sir Ganga Ram Hospital Lahore
6
Departmant of Pharmacology

Watim Dental College, Islamabad

Funding Source: NIL; Conflict of Interest: NIL
Received: July 04, 2018; Revised: October 23, 2018;
Accepted: November 11, 2018

Effects of Cichorium Intybus Seeds on NephrotoxicityJIIMC 2018 Vol. 13, No.4

184



proximal tubular cells are mainly exposed to damage 
by the drugs. Free radicals generated due to 
oxidative stress causes mitochondrial damage in 
tubules resulting in cytotoxicity. Inflammation in 
glomerulus and tubules is induced by nephrotoxic 
drugs which ultimately result in fibroses of renal 

 8
tissue.  
From ancient days medicinal plants have been widely 
used as a source of medication providing a cheap 
source of treatment for a majority of world's 
population. Herbal plants have got nephroprotective 

9
effect.  Several plants along with their extracts have 
got protective effect against nephrotoxic drugs due 
to their anti-oxidant, anti-inflammatory and diuretic 

10 
properties. Cichorium intybus , generally called 
chicory ( Kasni ) is vertical  woody perennial plant 
distributed in Europe, Mediterranean region, 

11 
Northern Asia and Pakistan. All parts of Cichorium 
intybus is  known to have antimicrobial, 
gastroprotective, hepatoprotective, anti-diabetic, 
antimalarial, anti-inflammatory and anti-oxidant 
effects due to the presence of phytocompounds like 

12,13 
polyphenols , alkaloids, flavonoids and tannins.
Previous studies have shown anti-oxidant effects of 
either Aqeous or methanolic extracts of Cichorium 
intybus roots, leaves and stem against gentamicin 
induced nephrotoxicity or hepatotoxicity in animal 

10 
model. Evaluation of both aqueous and methanolic 
extracts of Cichorium intybus seeds against 
gentamicin induced nephrotoxicity in literature is 
lacking. Present study was designed to evaluate the 
effect of both aqueous and methanolic extracts of 
Cichoriumintybus seeds against gentamicin induced 
nephrotoxicity in rat's model.

Materials and Methods
This experimental randomized control trial was 

th st
conducted from 15   January 2017 to 31  March 

2017 in the department of Pharmacology and 

Therapeutics, Islamic International Medical College, 

Rawalpindi in collaboration with the Riphah Institute 

of Pharmaceutical Sciences and Animal house at 

National Institute of Health (NIH), Islamabad. Forty 

healthy adult male Sprague Dawly rats were included 

through simple random sampling and the study was 

conducted after approval from Ethical Review 

Committee of Islamic International Medical College, 

Riphah International University (RIU), Islamabad.
Rats weighing between 300-350 grams with normal 

Serum urea and Creatinine levels were included in 
the study. Rats were first allowed to get acclimatized 
for one week in the NIH Animal house in 50-70% 

o
humidity at a room temperature of 24+2 C with a 12 

 14 
hour light and dark cycle. Rats were randomly 
divided into four groups of ten rats each (n=10). 
Group A was normal control, given diet and water ad 
libitum for ten days. Group B was administered 
Gentamicin at 80 mg/kg intraperitoneally once daily 
for ten days. Group C was administered Gentamicin 
80mg/kg intraperitoneally along with Aqueous 
extract of Cichorium intybus seeds orally mixed in 
2ml distal water through gavage tube at a dose of 

10 
500mg/kg once daily for ten days. Group D was 
administered Gentamicin 80mg/kg body weight 
once daily intraperitoneally along with Methanolic 
extract 500mg/kg orally mixed in 2ml distilled water 

15
through gavage tube once daily for ten days.
Dried seeds of Cichorium intybus were purchased 
and authenticated from herbarium department of 
National Agriculture and Research Centre (NARC) 
Islamabad. Aqueous extract of Cichoriumintybus 
seeds was prepared at RIPS, Islamabad by using fine 
homogenized powder of chicory seeds which were 
mixed with distilled water, the whole solution was 
boiled for 2 hours and after cooling was filtered 
through filter paper what man no 3. The aqueous 
extract was formed by using vacuum rotary 

16 
evaporator and was frozen dried. To obtain the 
methanolic extract, 1 kg of Cichorium Intybus seeds 
powder was soaked in 8 L of methanol (95%) for 24 h. 
The extract was filtered using what man paper No 3 
and the filtrate was evaporated (78 °C) to dryness 
using a rotary evaporator; 25.64 g of dried 
methanolic extract were obtained giving an 
extraction yield of 2.56% (w/w based on the dried 

17
starting weight).  Sampling at day 0 and day 6 were 

th
done via lateral tail vein. Sampling at day 11  was 
done through cardiac puncture. Biochemical analysis 
of serum urea and serum creatinine was estimated 
through commercially available kits by Merk and 
auto analyzer Micro lab 300 on photometric system.
Statistical analysis of collected parametric data was 
done by using SPSS version 21 and Mean ± Standard 
Error of Mean was calculated. One way ANOVA and 
Post Hoc Tuckey tests were applied to compare the 
mean difference between control and rest of the 
groups and mean difference in between the groups. 

Effects of Cichorium Intybus Seeds on NephrotoxicityJIIMC 2018 Vol. 13, No.4

185



p value of <0.05 was considered statistically 
significant.

Results
Mean initial Serum Urea for Group A (Normal Control 
Group) was 23.70 ± 1.521 mg/dl. Mean initial Serum 
Urea for Group B (Disease Control Group) was 24.00 
± 1.732 mg/dl. Mean initial Serum Urea for Group C 
(Aqueous extract Experimental group 1) was 25.56 ± 
1.744 mg/dL. Mean initial Serum Urea for Group D 
(Methanolic extract Experimental group 2) was 
22.10 ± 1.912 mg/dl. There was no significant 
difference in the serum urea levels among these 
groups on day 0 (p Value .589). Serum Urea on day 6 
for Group A (Normal Control Group) was 24.70 ± 
1.633 mg/dl, for Group B (Disease Control Group) 
was 83.30 ± 2.119 mg/dl, for Group C (Aqueous 
extract Experimental group 1) was 67.10 ± 1.703 
m g / d l &  fo r  G ro u p  D  ( M e t h a n o l i c  ex t ra c t  
Experimental group 2) was 84.00 ± 1.592 mg/dl. 
There was significant difference in the serum urea 
levels among these groups on day 6 (p Value .000). 
This suggests that nephrotoxicity was induced by 
Gentamicin.
Mean Serum Urea on day 11 for Group A (Normal 
Control Group) was 24.70 ± 5.165 mg/dl , for Group B 
(Disease Control Group) was 97.90 ±4.332 mg/dl, for 
Group C (Aqueous extract Experimental group 1 ) 
was 51.30 5.755 mg/dl& for Group D (Methanolic 
extract Experimental group 2) was 73.60mg/dl 
±4.695. There was significant difference in the values 
of group C & D (p Value .000). This suggests that 
nephrotoxicity in Group C (Aqueous Extract Treated 
Group) was improved more than Group D 
(Methanolic Extract Treated Group).
Mean initial Serum Creatinine for Group A (Normal 
Control Group) was 0.750 ± 0.0654 mg/dl. Mean 
initial Serum Creatinine for Group B (Disease Control 
Group) was 0.740 ± 0.0562 mg/dl. Mean initial 
Serum Creatinine for Group C (Aqueous extract 
Experimental group 1) was 0.770 ± 0.0559 mg/dL. 
Mean initial Serum Creatinine for Group D 
(Methanolic extract Experimental group 2) was 
0.770 ± 0.0667 mg/dL. There was no significant 
difference in the serum Creatinine levels among 
these groups on day 0. (p Value .980). 
Mean Serum Creatinine on day 6 for Group A 
(Normal Control Group) was 0.750 ± 0.0619 mg/dL, 
for Group B (Disease Control Group) was 2.490 ± 

0.0849 mg/dl, for Group C (Aqueous extract 
Experimental group 1) was 1.450 ± 0.0601 mg/dl& 
for Group D (Methanolic extract Experimental group 
2) was 1.730 ± 0.0517 mg/dl. There was significant 
difference in the serum Creatinine levels among 
these groups on day 6 (p Value .000). This suggests 
that nephrotoxicity was induced by Gentamicin.
Mean Serum Creatinine on day 11 for Group A 
(Normal Control Group) was 0.750, ± 0.0619 mg/dl, 
for Group B (Disease Control Group) was 2.490, ± 
0.0849 mg/dl, for Group C (Aqueous extract 
Experimental group 1 ) was 1.450  0.0601 mg/dl& for 
Group D (Methanolic extract Experimental group 2) 
was 1.730 ± 0.0517 mg/dl. There was significant 
difference in the values of group C & D (p Value .024). 
This suggests that nephrotoxicity in Group C 
(Aqueous Extract Treated Group) was improved 
more than Group D (Methanolic Extract Treated 
Group).

Table I: Comparison of Serum Urea and Crea�nine Levels
Among Groups By ANOVA at Day 0

Table II: Comparison of Serum Urea and Crea�nine Levels
Among Groups By ANOVA at Day 06 

Table III: Comparison of Serum Urea and Crea�nine 
Levels Among Groups By ANOVA at Day 11

Effects of Cichorium Intybus Seeds on NephrotoxicityJIIMC 2018 Vol. 13, No.4

186



Discussion
Gentamicin is widely used to induce nephrotoxicity 
in animal models for research purposes and has been 
stated by numerous researchers for analyzing 
nephroprotective effects of natural plants. Key 
phenomenon elaborating the GM induced 
nephrotoxicity is damage by reactive oxygen 

1 8  
species. Review of literature showed that 
production of ROS lead to lipid peroxidation causing 
necrosis and denaturation of proteins. Various 
studies have concluded that the nephroprotective 
effects of natural plants are due to the presence of 

19,20
antioxidants.
The current study was designed to investigate the 
protective effects of Aqueous and Methanolic 
extracts of Cichorium Intybus seeds in Gentamicin 

induced nephrotoxicity in rats via biochemical 
parameters. It was found that Gentamicin induced 
nephrotoxic changes were improved by both 
Aqueous and Methanolic extracts of Cichorium 
Intybus seeds but aqueous extract showed better 
nephroprotective effects against Gentamicin 
induced nephrotoxicity. In present study, GM 
induced nephrotoxicity is indicated by raised level of 
serum creatinine and urea as concluded previously 

21 
by Nitha and Janardhanan.  The process glomerular 
filtration causes the accumulation of gentamycin in 
the proximal tubules of kidney. GM Produced ROS 
causes oxidative stress and tubular injury and is 
indicated by raised serum creatinine and urea.
Our study in accordance with findings of study by 
Tanweer khaliq who induced Nephrotoxicity in 
Albino rabbits with Gentamicin Intraperitoneally in a 
dose of 80 mg/kg and observed raised levels of 

10 
biochemical markers. Similarly our study is in 
correlation with another study performed by V. 
Chinnapa Reddy showing the nephrotoxic effects of 
Gentamicin in Rats which were indicated by raised 

 16
levels of serum urea and creatinine.  
Current study is consistent with the study of Tanweer 
Khaliq who compared the nephroprotective effect of 
aqueous extract of Cichorium Intybus seeds against 
Gentamicin induced nephrotoxicity in which he used 
aqueous extract of Cichorium Intybus seeds in doses 
of 250 mg/kg administered via gavage tube. His 
study also evidenced the nephroprotection due to 
presumed antioxidant properties of Cichorium 

 10
Intybus seeds.
Review of literature shows that studies have been 
done on exploring nephroprotective effect of 
Cichorium intybus in combination with other herbal 

10  
extracts and medical compounds like silymarin. No 
individual study was done on aqueous and 
methanolic extract of Cichorium intybus seed 
indicating dosage use regarding submaximal, ceiling 
effect and toxicity. Current study shows the 
nephroprotective effect of both aqueous and 
methanolic extract of Cichorium intybus seeds. 
Further studies are needed to elaborate the 
mechanism of higher response of methanolic extract 
as compare to aqueous extract of Cichorium intybus 
seeds for reducing elevated renal markers. In 
addition administration through different routes and 
higher dosage can be tried to see the effect.

Table IV: Mul�ple Comparison of Serum Urea and 
Crea�nine levels Among all Groups By Post Hock Tuckey
Test

Effects of Cichorium Intybus Seeds on NephrotoxicityJIIMC 2018 Vol. 13, No.4

187



Conclusion
Aqueous and Methanolic extracts of Cichorium 
Intybus seeds exhibited protective effects against 
Gentamicin induced nephrotoxicity in rats. Aqueous 
extract Cichorium intybus seeds have exerted 
greater nephroprotective effect in comparison to 
Methanolic extract.

REFERENCES
1. Hussein O, Germoush M, Mahmoud A. Ruta graveolens 

P r o t e c t s  A g a i n s t  I s o n i a z i d / R i f a m p i c i n - I n d u c e d  
Nephrotoxicity through Modulation of Oxidative Stress and 
Inflammation. Glob J Biotechnol Biomater Sci: 2016;50 
(1):17-22.

2. Singh RK, Gautam RK, Karchuli M. Nephrotoxicity: an 
overview. Journal of Biomedical and Pharmaceutical 
Research: 2014;3(4):41-7.

3. Kandasamy K, Chuah JKC, Su R, Huang P, Eng KG, Xiong S, et 
al. Prediction of drug-induced nephrotoxicity and injury 
mechanisms with human induced pluripotent stem cell-
derived cells and machine learning methods. Scientific 
reports: 2015;5(2): 45-55.

4. Shi K, Caldwell SJ, Fong DH, Berghuis AM. Prospects for 
circumventing aminoglycoside kinase mediated antibiotic 
resistance. Frontiers in cellular and infection microbiology. 
2013;3:22-28.

5. Katzung BG, Masters SB, Trevor AJ. Basic & clinical 
pharmacolo gy. 12th ed: McGraw-Hill Medical New York; 
2016.

6. Filazi A, Sireli UT, Pehlivanlar-Onen S, Cadirci O, Aksoy A. 
Comparative Pharmacokinetics of Gentamicin in Laying 
Hens. Kafkas Üniversitesi Veteriner Fakültesi Dergisi: 
2013;19(3):495-98.

7. Ramirez MS, Tolmasky ME. Aminoglycoside modifying 
enzymes. Drug Resistance Updates: 2010;13(6):151-71.

8. Kim S-Y, Moon A-R. Drug-induced nephrotoxicity and its 
b i o m a r k e r s .  B i o m o l e c u l e s  a n d  T h e r a p e u t i c s :  
2012;20(3):268-72.

9. Maharana L, dadhich op. Review on role of herbal drug in 
the prevention and management of kidney disease. 
Ayushdhara:2016;1(1): 15-20.

10. Khaliq T, Mumtaz F, Rahman Z, Javed I, Iftikhar A. 
Nephroprotective potential of Rosa damascena mill 
flowers, Cichorium intybus linn roots and their mixtures on 
gentamicininduced toxicity in albino rabbits. Pak Vet J: 

2015;35:43-47.
11. Hozayen WG, El-Desouky MA, Soliman HA, Ahmed RR, 

Khaliefa AK. Antiosteoporotic effect of Petroselinum 
crispum, Ocimum basilicum and Cichorium intybus L. in 
glucocorticoid-induced osteoporosis in rats. BMC 
complementary and alternative medicine: 2016;16(1):165-
169.

12. Saxena R, Sulakhiya KB, Rathore M. Cichorium intibus Linn.: 
a review of pharmacological profile. International Journal of 
Current Pharmaceutical Research: 2014;6(4):11-15.

13. Shad M, Nawaz H, Rehman T, Ikram N. Determination of 
some biochemicals, phytochemicals and antioxidant 
properties of different parts of Cichorium intybus L.: A 
comparative study: J Anim Plant Sci. 2013;23(4):1060-66.

14.   Al-Yahya M, Mothana R, Al-Said M, Al-Dosari M, Al-
Sohaibani M, Parvez M, et al. Protective effect of citrus 
medica'otroj'extract on gentamicin-induced nephrotoxicity 
and oxidative damage in rat kidney. Digest Journal of 
Nanomaterials &Biostructures (DJNB): 2015;10(1):15-19. 

15. Reddy VC, Amulya V, Lakshmi C, Reddy K, Praveen D, 
Pratima D, et al. Effect of Simvastatin in gentamicin induced 
nephrotoxicity in albino rats. Asian J Pharm Clin Res: 
2012;5:36-40.

16. Ali A, Jehangir A, Ayub F. Hepatoprotective Effect of 
Aqueous Extract of Cichorium Intybus Roots on Isoniazid 
Induced Hepatotoxicity. JIIMC. 2016; 11(3): 99-102.

17. Watcho P, Watio HM, Wankeu-Nya M, Ngadjui E, Defo PD, 
Nkeng-Efouet PA, et al. Androgenic effects of aqueous and 
methanolic extracts of Ficus asperifolia in male Wistar rats. 
B M C  co m p l e m e nta r y  a n d  a l te r n at i ve  m e d i c i n e :  
2017;17(1):42-46.

18. Weber EJ, Himmelfarb J, Kelly EJ. Concise Review: Current 
and Emerging Biomarkers of Nephrotoxicity. Current 
Opinion in Toxicology: 2017; 4: 16-21.

19.  Chen Q, Cui Y, Ding G, Jia Z, Zhang Y, Zhang A, et al. PEA3 
protects against gentamicin nephrotoxicity: role of 
mitochondrial dysfunction. American journal of 
translational research: 2017; 9(5):2153-58.

20. Duscha S, Boukari H, Shcherbakov D, Salian S, Silva S, 
Kendall A, et al. Identification and evaluation of improved 
4ʹ-O-(alkyl) 4, 5-disubstituted 2-deoxystreptamines as next-
g e n e r a t i o n  a m i n o g l y c o s i d e  a n t i b i o t i c s .  M B i o :  
2014;5(5):e01827-14.

21.  Nitha B, Janardhanan KK. Aqueous-ethanolic extract of 
morel mushroom mycelium Morchella esculenta, protects 
cisplatin and gentamicin induced nephrotoxicity in mice. 
Food and Chemical Toxicology: 2008; 46(9): 3193-99.

Effects of Cichorium Intybus Seeds on NephrotoxicityJIIMC 2018 Vol. 13, No.4

188