academic sciences · 2018-04-23 · research article hepatportective activity of elettaria...

3
Research Article HEPATPORTECTIVE ACTIVITY OF ELETTARIA CARDAMOMUM AGAINST PARACETAMOL INDUCED HEPATOTOXICITY NIMMY CHACKO*, AMBILY THOMAS, C S SHASTRY, PRERANA SHETTY *Department of Pharmacology, NGSM Institute of Pharmaceutical Sciences, Paneer, Deralakatte, Manglore, Karnataka, India. Email: [email protected] Received: 15 Mar 2012, Revised and Accepted: 21 Apr 2012 ABSTRACT Aim of study: To evaluate the hepatoprotective activity of fruits of Elettaria cardamomum extract against Paracetamol induced hepatotoxicity. Materials and methods: Methanolic extract at doses of 100, 200 and 400 mg/kg were administered orally once daily for 7 days. The hepatoprotective activity was assessed using various biochemical parameters like alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and total bilirubin, along with histopathological studies of liver. Result: Elettaria cardamomum exhibited significant hepatoprotective activity by reversing bio‐chemical changes induced by paracetamol. Histology of the liver sections of the animals treated with the extracts showed the presence of almost normal hepatocytes, absence of necrosis and fatty infiltration, which further evidenced the hepatoprotective activity. Conclusion: Methanolic extract of fruits of Elettaria cardamomum has liver protective effect against paracetamol induced hepatotoxicity. Further studies are needed to isolate and characterize the active principles and to find out the mechanism responsible for its hepatoprotective activity. Keywords: Antihepatotoxic, Paracetamol, Elettaria cardamomum, Liver injury. INTRODUCTION Liver is one of the largest organs in human body and the chief site for intense metabolism and excretion. So it has a surprising role in the maintenance, performance and regulating homeostasis of the body 1 . The major functions of the liver are carbohydrate, protein and fat metabolism, detoxification, secretion of bile and storage of vitamins. Thus, to maintain a healthy liver is a crucial factor for overall health and well being. But it is continuously and variedly exposed to environmental toxins, prescribed and over‐the‐counter drugs which can eventually lead to various liver ailments like hepatitis, cirrhosis and alcoholic liver disease 2 . Thus liver diseases are some of the fatal disease in the world today. They pose a serious challenge to international public health. Modern medicines have little to offer for alleviation of hepatic diseases and it is chiefly the plant based preparations which are employed for the treatment of liver disorders. But there is not much drug available for the treatment of liver disorders 3 . In absence of a reliable liver protective drug in the modern system of medicine, a number of medicinal preparations in Ayurveda, the Indian system of medicine, are recommended for the treatment of liver disorders. Natural remedies from medicinal plants are considered to be effective and safe alternative treatments for hepatotoxicity 4 . In view of this, the present study is undertaken to investigate the hepatoprotective activity of Elettaria cardamomum fruits against paracetamol induced hepatotoxicity in rats. MATERIALS AND METHODS Plant material and extraction The dried fruits of Elettaria cardamomum were purchased from the local markets of Kerala in the month of June 2010. The fruits were identified and authenticated by Dr. Jomy Augustine, Head of the Department of Botany, St. Thomas College, Pala, Kerala. After cleaning of adulterant material, the fruits were ground with an electric grinder into a coarse powder. About 500 g of ground material was soaked in methanol (70%) at room temperature (23–25 C) for 3 days with occasional shaking. It was filtered through a muslin cloth and then through a Whatman qualitative grade 1 filter paper. This procedure was repeated twice and the combined filtrates were evaporated on rotary evaporator under reduced pressure (−760 mmHg) to a thick, semi‐solid pasty mass of dark brown color; i.e. the crude extract yielding approximately 4.9%. Experimental animals Albino Wistar rats of either sex (150‐200 g) were obtained from the laboratory of NGSM Institute of Pharmaceutical Sciences, Deralakatte, Mangalore. Four animals were housed in a cage in a climate‐controlled room under standard conditions with 12:12 hour’s light/dark cycles and free access to water and food. Animals were brought in to the laboratory for at least a week before experimental testing commenced. Each experimental group of rats were randomly chosen and used only once. All the experiments were performed within the guidelines of the institutional animal ethical committee of KSHEMA. Deralakatte, Mangalore (KSHEMA/AEC/40/2010). Hepatoprotective Activity Animals were divided into six groups of 6 animals each. The first group received saline 1 ml/kg for one week (control). The group II received saline 1 ml/kg for one week (positive control). The group III received silymarin (25 mg/kg p.o.). The group IV, V and VI received 100, 200 and 400 mg/kg of extract respectively once a day for seven days. On the fifth day, after the administration of the respective treatments, all the animals of groups II, III, IV, V and VI were administered with paracetamol 2 g/kg orally. On the seventh day after 2 h of respective treatments, the blood samples were collected from all groups of rats by puncturing the retro‐orbital plexus. Serum was separated by centrifugation at 2500 rpm at 37 o C for 15 min and analyzed for various biochemical parameters. Liver was dissected out and used for histopathological studies 5 . Assessment of liver functions Biochemical parameters, such as Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Alkaline phosphatase (ALP) and Total bilirubin (TBIL) were estimated using the assay kit (AGAPPE Diagnostics Ltd, Ernakulam, Kerala). International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 4, Issue 3, 2012 A A c c a a d d e e m mi i c c S S c c i i e e n n c c e e s s

Upload: others

Post on 20-Feb-2020

4 views

Category:

Documents


0 download

TRANSCRIPT

Page 1: Academic Sciences · 2018-04-23 · Research Article HEPATPORTECTIVE ACTIVITY OF ELETTARIA CARDAMOMUM AGAINST PARACETAMOL INDUCED HEPATOTOXICITY NIMMY CHACKO*, AMBILY THOMAS, C S

Research Article 

HEPATPORTECTIVE ACTIVITY OF ELETTARIA CARDAMOMUM AGAINST PARACETAMOL INDUCED HEPATOTOXICITY 

 

NIMMY CHACKO*, AMBILY THOMAS, C S SHASTRY, PRERANA SHETTY 

*Department of Pharmacology, NGSM Institute of Pharmaceutical Sciences, Paneer, Deralakatte, Manglore, Karnataka, India.                              Email: [email protected] 

Received: 15 Mar 2012, Revised and Accepted: 21 Apr 2012 

ABSTRACT 

Aim of study: To evaluate the hepatoprotective activity of fruits of Elettaria cardamomum extract against Paracetamol induced hepatotoxicity. 

Materials and methods: Methanolic extract at doses of 100, 200 and 400 mg/kg were administered orally once daily  for 7 days. The hepatoprotective activity was assessed using various biochemical parameters like alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and total bilirubin, along with histopathological studies of liver. 

Result: Elettaria cardamomum exhibited significant hepatoprotective activity by reversing bio‐chemical  changes  induced by paracetamol. Histology of the liver sections of the animals treated with the extracts showed the presence of almost normal hepatocytes, absence of necrosis and fatty infiltration, which further evidenced the hepatoprotective activity. 

Conclusion: Methanolic extract of fruits of Elettaria cardamomum has liver protective effect against paracetamol induced hepatotoxicity. Further studies are needed to isolate and characterize the active principles and to find out the mechanism responsible for its hepatoprotective activity. 

Keywords: Antihepatotoxic, Paracetamol, Elettaria cardamomum, Liver injury. 

 

INTRODUCTION  

Liver is one of the largest organs in human body and the chief site for intense  metabolism  and  excretion.  So  it  has  a  surprising  role  in  the maintenance,  performance  and  regulating  homeostasis  of  the  body 1. The  major  functions  of  the  liver  are  carbohydrate,  protein  and  fat metabolism,  detoxification,  secretion  of  bile  and  storage  of  vitamins. Thus,  to maintain  a  healthy  liver  is  a  crucial  factor  for  overall  health and  well  being.  But  it  is  continuously  and  variedly  exposed  to environmental  toxins,  prescribed  and  over‐the‐counter  drugs  which can  eventually  lead  to  various  liver  ailments  like  hepatitis,  cirrhosis and alcoholic  liver disease 2. Thus  liver diseases are  some of  the  fatal disease  in  the  world  today.  They  pose  a  serious  challenge  to international  public  health. Modern medicines  have  little  to  offer  for alleviation  of  hepatic  diseases  and  it  is  chiefly  the  plant  based preparations which are employed for the treatment of liver disorders. But  there  is  not  much  drug  available  for  the  treatment  of  liver disorders 3. In absence of a reliable liver protective drug in the modern system of medicine, a number of medicinal preparations in Ayurveda, the Indian system of medicine, are recommended for the treatment of liver disorders. Natural remedies from medicinal plants are considered to  be  effective  and  safe  alternative  treatments  for  hepatotoxicity 4.  In view  of  this,  the  present  study  is  undertaken  to  investigate  the hepatoprotective  activity  of  Elettaria  cardamomum  fruits  against paracetamol induced hepatotoxicity in rats.  

MATERIALS AND METHODS 

Plant material and extraction 

The  dried  fruits  of  Elettaria cardamomum  were  purchased  from  the local  markets  of  Kerala  in  the  month  of  June  2010.  The  fruits  were identified  and  authenticated  by  Dr.  Jomy  Augustine,  Head  of  the Department of Botany, St. Thomas College, Pala, Kerala. After cleaning of adulterant material, the fruits were ground with an electric grinder into a  coarse powder. About 500 g of  ground material was  soaked  in methanol  (70%)  at  room  temperature  (23–25  C)  for  3  days  with occasional  shaking.  It  was  filtered  through  a  muslin  cloth  and  then through  a Whatman  qualitative  grade  1  filter  paper.  This  procedure 

was  repeated  twice  and  the  combined  filtrates  were  evaporated  on rotary  evaporator  under  reduced  pressure  (−760 mmHg)  to  a  thick, semi‐solid  pasty  mass  of  dark  brown  color;  i.e.  the  crude  extract yielding approximately 4.9%. 

Experimental animals 

Albino Wistar  rats  of  either  sex  (150‐200  g) were  obtained  from  the laboratory of NGSM Institute of Pharmaceutical Sciences, Deralakatte, Mangalore. Four animals were housed in a cage in a climate‐controlled room  under  standard  conditions  with  12:12  hour’s  light/dark  cycles and  free  access  to  water  and  food.  Animals  were  brought  in  to  the laboratory for at least a week before experimental testing commenced. Each experimental group of rats were randomly chosen and used only once. All the experiments were performed within the guidelines of the institutional  animal  ethical  committee  of  KSHEMA.  Deralakatte, Mangalore (KSHEMA/AEC/40/2010). 

Hepatoprotective Activity 

Animals were divided into six groups of 6 animals each. The first group received saline 1 ml/kg for one week (control). The group II received saline 1 ml/kg for one week (positive control). The group III received silymarin  (25 mg/kg p.o.).  The  group  IV,  V  and VI  received 100,  200 and 400 mg/kg of  extract  respectively once a day  for  seven days. On the fifth day, after the administration of  the respective treatments, all the  animals  of  groups  II,  III,  IV,  V  and  VI  were  administered  with paracetamol 2 g/kg orally. On the seventh day after 2 h of respective treatments, the blood samples were collected from all groups of rats by puncturing  the  retro‐orbital  plexus.  Serum  was  separated  by centrifugation at 2500 rpm at 37oC for 15 min and analyzed for various biochemical  parameters.  Liver  was  dissected  out  and  used  for histopathological studies 5. 

 Assessment of liver functions 

Biochemical  parameters,  such  as  Alanine  aminotransferase  (ALT), Aspartate  aminotransferase  (AST),  Alkaline  phosphatase  (ALP)  and Total  bilirubin  (TBIL)  were  estimated  using  the  assay  kit  (AGAPPE Diagnostics Ltd, Ernakulam, Kerala). 

International Journal of Pharmacy and Pharmaceutical Sciences

ISSN- 0975-1491 Vol 4, Issue 3, 2012

AAccaaddeemmiicc SScciieenncceess  

Page 2: Academic Sciences · 2018-04-23 · Research Article HEPATPORTECTIVE ACTIVITY OF ELETTARIA CARDAMOMUM AGAINST PARACETAMOL INDUCED HEPATOTOXICITY NIMMY CHACKO*, AMBILY THOMAS, C S

 

H

Tseap

S

TPfos

GI IIII

IV

V

V

V

H

 

 

Histopathology 

The  liver  tissueolution,  dehydrembedded  in  paand  then  staiphotomicroscopi

Statistical Analy

The data were exP  <  0.05  betweeollowed  by  Dunoftware. 

Tabl

Group No. 

I II 

VI 

Values are expres

a ‐ significant (p<0

Histopathologica

 

 

 

  was  dissectedated  in  ethanol raffin  wax.  5–6mned  with  hemic observation.  

ysis 

xpressed as meaen  the  groups  wnnett’s  multiple 

e 1: Effect of me

Groups (n=6) Control  Pcml Treated Pcml + Silymari(25 mg/kg bodyPcml + MEC  (100 mg/kg bodPcml + MEC  (200 mg/kg bodPcml + MEC  (400 mg/kg bod

sed as mean ± SE

0.05) compared t

al studies under 

Fig. 1: Live

Fig. 3: Liver t

  out  and  fixed (50–100%)  cleamm  thick  sectiomatoxylin  and 

an ± S.E.M. Statiswere  analyzed  bycomparison  tes

thanolic extract

 

in  y wt.) 

dy wt.) 

dy wt.) 

dy wt.) 

EM, n=6 rats in ea

to control, b ‐ sign

 light microscop

er tissue: Contro

 tissue: Silymarin

in  10%  formaared  in  xylene  aons  were  prepar

eosin  dye 

stical differencesy  one‐way  ANOst  using  SPSS  1

ts of fruits of Ele

ALT (IU/L) 89.0±1.15 288.3±1.47a 97.83±1.66b 

195.2±1.13 

134.5±1.12b 

112.8±2.97b 

ch group 

nificant (p<0.05) 

py 

ol Group  

n Treated  

Chacko et al. 

alin and red for 

s at OVA 5.0 

RE

Thebe fou

Ser

Admas Preexhhepextby 

 

ttaria cardamom

AST (IU/L110.2±1.3383.8±3.3123.8±2.7

285.2±1.3

182.5±0.7

162.0±1.1

compared to Pcm

 

 

 

Fig

SULTS  

e methanolic extrnontoxic when  aund to be safe up t

rum parameters

ministration of pevidenced  by 

etreatment of ratshibited  markedpatotoxicity, (shotract of Elettaria cthe standard, sily

mum on paracet

L)  ALP (5  142.08a  389.71b  152.3

0  294.8

6b  194.0

5b  169.0

ml 

Fig. 2: Liver

g. 4: Liver tissue:

Int J Pharm P

ract of fruits of Eladministered  orato 2 g/kg body w

s determination 

aracetamol to rathe  altered  s

s with methanolid  protection own in Table 1). Tcardamomum weymarin.  

amol induced he

  IU/L)  T±2.01  0.±2.49a  2.±2.12b  0.

±1.27  1.

±1.18b  1.

±1.75b  0.

r tissue: Paracet

: (100mg/kg ext

Pharm Sci, Vol 4,

lettaria cardamomally  to  rats  and  itt. 

 

ts caused significserum  biochemc extract of Elettaagainst  parac

The effects producere comparable w

epatotoxicity in 

otal Bilirubin (m.74±0.02 .23±0.02a .85±0.02b 

.74±0.01 

.15±0.01b 

.88±0.01b 

tamol Treated G

tract) Group Tre

, Issue 3, 611­613

61

mum was found tts  LD50  value wa

cant liver damagemical  parametersaria cardamomumetamol  induceced by methanoliwith that produce

 rats. 

mg/dL) 

 

roup 

 

eated Group 

to as 

e, s. m d ic d 

Page 3: Academic Sciences · 2018-04-23 · Research Article HEPATPORTECTIVE ACTIVITY OF ELETTARIA CARDAMOMUM AGAINST PARACETAMOL INDUCED HEPATOTOXICITY NIMMY CHACKO*, AMBILY THOMAS, C S

 

 

D

Tamm

AAamppe6,

h

PObpsoceoth

Wchdah 

  Fig. 5: L

DISCUSSION AND

The present studyactivity  of methanmethanolic extracmortality up to 20

Administration ofALP and Total Bila  toxic  electrophimethanolic  extraprevent  the  elevparacetamol.  Theeffects  on  cytochr,7.  These  biochhistopathological 

Paracetamol is noOnly  5%  of  tbenzoquineimine.paracetamol,  theaturated and henoxidized  to  highlcytochrome‐450 electron reductionof cellular membrhe tissue damage

With aid of enzymcan conclude that have  hepatoprotdamage.  Further active  principles hepatoprotective 

Liver tissue: (20

D CONCLUSION 

y was under takenolic  extract  of  fct did not show an000 mg/kg dose. 

f paracetamol eleirubin significantile,  N‐acetyl‐p‐bect  of  fruits  of  Evation  of  ALT, se  biochemical  erome P450  and/hemical  findingsstudies. 

ormally eliminatethe  paracetamo.  However,  upone  sulfation  andnce, higher percely  reactive  N‐aceenzymes.  A  semn of NAPQI,  can rane and increasee 8.  

me levels and histmethanolic extraective  activity studies  are  needand  to  find  out activity. 

00mg/kg extract

n to determine thfruits  of Elettariany sign and symp

evated the serumtly. This is due toenzoquine‐imine.Elettaria  cardamoAST,  ALP  and effects may  be  duor  promotion  of s  were  further 

ed mainly as sulfal  is  convertedn  administrationd  glucuronidatioentage of paracetetyl‐p‐benzoquinmiquinone  radicacovalently binds es the lipid perox

topathological stuact of fruits of Eleagainst  Paracetaded  to  isolate  anthe  mechanism 

t) Treated Group

he hepatoprotecta cardamomum.  Tptoms of toxicity a

m  levels of ALT, Ao its bioactivation  Pretreatment  womum  was  able Total  Bilirubin ue  to  the  inhibitoits  glucuronidatsubstantiated 

ate and glucuronid  into  N‐acetyln  of  toxic  doses on  routes  becotamol molecules eimine  (NAPQI) al,  obtained  by  oto macromolecuxidation resulting

udies of rat liver ettaria cardamomamol  induced  livnd  characterize responsible  for 

Chacko et al. 

 

p  F

    

tive The and 

AST, n to with to by ory tion by 

ide. l‐p‐of 

ome are by one ules g in 

we mum ver the its 

AC

ThePhathis

RE

1.

2.

3.

4.

5.

6.

7.

8.

9.

10.

Fig. 6: Liver tissu

KNOWLEDGEME

e  author  is  thanarmaceutical Scies study. 

FERENCES  

Ward  FM,  DalTherapeutics. NSharma A, Chasome  Indian Fitoterapia.199Karan M, Vasischirata  on  carPhytotherapy RChatterjee  TK.(P) Ltd.2000. Chattopadhyayactivity  of EthnopharmacWesley  GC,  BrUSA: Mosby YeGilman AG, Ralof TherapeuticDiadelis R,  JanPharmacol: EnShazia UsmaniActivity  Of 2010;2(3):101

. Arief  NurrochMargono,  SarHepatoprotectagainst carbon2010;2(3):45‐4

Int J Pharm P

ue: (400mg/kg e

ENT  

nkful  to  Nitte  Guences for providin

ly  MJ.  Hepatic  DNew York: Churchakraborti KK, Hanherbal  formulat91;62: 229‐235. sht K, Handa SS. Arbon  tetrachloridResearch.1999; 1.  Herbal  Options.

y  RR.  Possible Azardica  indi

cology. 2003; 89: rater  CC,  Alice  Rear Book; 1992;41ll TW, Nies AS, Tacs. 13th ed. Londonn NM,  Commandenviron Toxicol Phai,  Poonam kushwCalotropis  G

1‐3 hmad,  Arief  Rardjiman  and  Ntive  and  Antioxidn tetrachloride in48 

Pharm Sci, Vol 4,

 extract) Treated

ulabi  Shetty  Memng necessary faci

Disease  in  Clinichill Livingstone; 1nda SS. Anti‐hepations  as  compar

Antihepatotoxic ade  induced  hepa13: 24‐30. .  1st  ed.  Kolkata: 

mechanism  of ca  leaf  extra217. RJ.  Cloth’s  Medic1. aylor P. The Pharn: Mc Graw Hill; 1eur  ED, Nico PE. armacol Sect. 199waha.A  Study On Gigantea  Leave

ahman  Hakim, Nunung  Yuniartidant  activity  of  Hduced hepatic inj

, Issue 3, 611­613

61

 

d Group 

morial  Institute  oilities to carry ou

al  Pharmacy  an1999;195‐212. atotoxic activity ored  to  Silymarin

activity of Swertitotoxicity  in  rats

Books  and Allie

hepatoprotectivact.  Journal  o

cal  Pharmacology

rmacological Basi1992. Vermeulen.  Eur 95; 293: 301. Hepatoprotectivs  Extract.IJPPS

Supardjan  Amii.  Evaluation  oHexagamavunon‐jury in rats. IJPPS

of ut 

of n. 

a s. 

ve of 

y. 

is 

ve S. 

ir of 1 S.