Ancient Science of Life, Vol. 28, No.

4 (2009) Pages 32 - 35

A Study of Hepatoprotective activity of

Hedyotis corymbosa. Linn. in albino rats

Abstract: Hedyotis corymbosa Linn is spreading, suffruticose annual, belongs to family

Rubiaceae frequently met with in field through out India, usually during rainy season. This plant
Rajshekar Chimkode1 is used for their medicinal properties as a folk medicine to treat jaundice, mouth wash in toothache.
M.B Patil 2 Hence the present study was aimed to investigate the hepatoprotective activity of ethanolic extract
of Hedyotis corymbosa which was separated in to different fractions against carbon tetrachloride
Sunil Jalalpure 3
intoxification. The results indicate that, Intoxification with CCl4 increase the levels of SGOT
T.Y. Pasha1 and SGPT. The elevated levels of SGOT and SGPTwere significantly decreased by ether and
Sibaji sarkar1. butanol fractions at P<0.001 and butanone and ethanol at p < 0.05 , where as petroleum ether
and ethyl acetate did not shown any significant reduction in the level of SGOT and SGPT.
Key words: Hepatoprotective activity, Hedyotis corymbosa Linn. Ethanolic extract in
different fraction

Introduction activity [7, 8]. Some other plant of this genus have
The liver is the largest organ in the body weighing been reported to possess other activity like anti
1400-1600 gm in the males and 1200-1400 gm in oxidant, anti inflammatory. [9, 10] Hence the present
the females [1] and this organ also regulating study was aimed to investigate the
homeostasis in the body .Thus liver is expected not hepatoprotective activity of ethanolic extract of
only to perform physiological function , but it has Hedyotis corymbosa which was separated in to
to protect itself against the hazards of harmful different fractions against carbon tetrachloride
medicines and chemicals [2]. Inspite of the intoxification.
tremendous scientific advancement in the field of
hepatology in recent years, problem have been Material and Methods
added rather than solved [3]. The liver injury may Drugs and chemicals
take several forms and involve the hepatocytes, LIV-52 obtained commercially from Himalaya
likes vascular cells or bile ducts. The most Drug co. Bangalore, India. The kits for all
important diseases are: biliary obstruction, biochemical were purchased from Marck
metabolic lesions caused by genetic disorder or chemicals. The solvent and other chemicals used
exogenous substance, such as alcohol, were of analytical grade.
inflammation, especially caused by cirrhosis,
neoplasia[4,5]. Presently only a few Plant material and extracts
hepatoprotective drugs and that too from natural Hedyotis corymbosa. Linn was collected from
sources, are available for the treatment of liver local market at udupi and was authenticated by Dr.
disorders [6]. K.K Srinivasan, faculty, college of pharmaceutical
sciences Manipal (Karnataka).
Hedyotis corymbosa, linn is spreading ,
suffruticose annual, up to 15 inch high, belongs to 1
family Rubiaceae, known as Parpatah in sanskrit,, J.J.C trust sanchalit N.R Vekaria institute of
frequently met with in field throughout India, pharmacy and research center, Junagadh. Gujarat
usually during rainy season. The plant is used in -362001, India.
2
the treatment of jaundice. In Phillipines, the plant K.L.E. Society, s College of pharmacy, Ankola,
is boiled in water and the brew used as mouth wash Karnataka -581314. India.
3
in toothache and Hedyotis corymbosa. Linn also K.L.E. Society, s College of pharmacy, Belgaum,
have immuno competent activity and antitumour Karnataka 590010. India

pages 32 - 35
 ANCIENT SCIENCE OF LIFE
After authentication, the fresh Hedyotis
corymbosa were shade dried and then milled in to
coarse powder by a mechanical grinder and
powder was used for further studies. The air-dried
powder plant (3kg) of Hedyotis corymbosa was
exhaustively extracted with 95% ethanol in 12
batches in a soxhlet apparatus. The ethanolic
extract then concentrated under reduced pressure
and controlled temperature to a small volume (500
ml). This was then evaporated on water bath and
residue to dryness (210 g.). A 15 g. sample
reserved for preliminary pharmacological and
phytochemical studies. Rest of it was suspended
and dispersed in 500 ml distilled water and
extracted with equal volume ( thrice with 200 ml )
of petroleum ether ( 60-80 %) , solvent ether, ethyl
acetate, butanone, n-butanol. After that each
extract was washed with water and dried over
anhydrous sodium sulphate. Then extract was
concentrated to a small volume under reduced
pressure and evaporated to dryness.( yield-
petroleum ether-12 g. , solvent ether-10 g. , ethyl
acetate 11 g., butanone-12 g. , n- butanol -09 g., ).
After preliminary phytochemical tests, it was
found that petroleum ether extract do not contain
any sterol and triterpenoid, but solvent ether
extract contains alkaloids. It was also found that
ethyl acetate, butanone and n- butanol extract
contains coumarins. The 0.5 g. of each extract was
reserved for pharmacological activity.
Animals
Adult, albino mice (20-25 gram) and albino rats
(200-250 gram ) were obtained from animal
house, N.R Vekaria institute of pharmacy and
research center, Junagadh were used for the study .
The animals were maintained under basal
metabolic diet and environmental condition
throughout the experiments. They were housed in
acrylic cages under the standard laboratory
condition at 22-25 0 C for performing the
experiment. The animals were provided with Rat-
feed (Anti diet A) which was supplied by M/S.
Lipton India Ltd, Bangalore. Before conducting
the experiment, the animal ethical clearance was
obtained from Institutional animal ethics
committee (IAEC), approved by committee for the
purpose of control and supervision of Experiment
on Animals (CPCSEA)
LD 50 determination
An acute toxicity study was carried out for
pages 32 - 35
determination of LD 50 for different extract, in
albino mice of either six weights 20-25 g. The
animal were fasted over night prior to acute
experimental procedure and six groups of animals
containing 6 in each , received test extracts in the
dose , ranging from 500 to 3000 mg /kg , body
weight /orally , each dose bearing ratio of ½ from
its antecedent dose . All the extracts were
administered orally as suspension in propylene
glycol. The method of miller and Trainer 1954 was
adopted for determination of LD 50 [11].
Hepatoprotective activity
Modified method of Chandra et. al has been used
in this study. Propylene glycol, which is found to
be safe, non toxic and non interfering solvent has
been used to prepare the solution of the drug.
Animals were administered carbon tetrachloride
(0.4 ml/kg) I.P for five days to induce
hepatotoxicity. Marked increased in the serum
levels of SGOT and SGPT were taken as
indication of hepatotoxicity. A group of 48 animals
each weighing approximately 200 to 250 gms
were used for recording enzymic levels and
histopathology during the evaluation. Normal
levels for the serum enzymes SGOT and SGPT
were determined by withdrawing blood samples
from unanaesthetised animals by direct heart
puncture. Mean value of SGOT and SGPT were
noted. Six animals were sacrificed and then liver
samples were subjected to histopathological
analysis .The animals left out from the first stet
were administered carbon tetrachloride at a dose
of 0.4 ml/kg. I. P daily for five days. On the sixth
day, six animals were taken an enzymic levels
were determined. mean value of SGOT and SGPT
were finally noted .Then the animals were
sacrificed and hepatotoxicity was assessed
histopathologically. Administration of carbon
tetra chloride was stopped on the sixth day and
administration of drug extracts and LIV-52 in
propylene glycol was started , this was continued
for five days. On the 11th days the serum levels
were noted and the mean value was recorded.
Histopathologies of liver samples from animals
were assessed.
Results:
The ethanolic and petroleum ether extract did not
shown any toxicity and mortality even at the dose
of 3000 mg/kg by intraperitoneal or oral route .
Hence the orbitory dose 500 mg/kg body wt.
 A STUDY OF HEPATOPROTECTIVE ACTIVITY OF HEDYOTIS CORYMBOSA. LINN. IN ALBINO RATS

Table 1
Effect of LIV -52 and various test extracts on
CCL4 induced hepatototoxicity in rats

S. no Compounds SGOT SGPT

Mean  SE Mean  SE
1 Normal 245.50  6.60 54.50  2.21
2 CCL4 induced 589.66  5.200 213.50  2.87
3 Ethanol 417.83  2.56** 112.00  1.87**
4 Petroleum ether 437.50  13.12 123.66  3.15
5 Ether 353.66  1.56*** 72.83  0.90***
6 Ethyl acetate 435.00  10.30 114.16  2.42
7 Butanone 416.33  3.02** 119.83  2.66**
8 Butanol 353.33  3.54*** 74.00  3.17***
9 LIV-52 346.16  1.55*** 70.33  1.05***

No. of the animals in each group =6

*** Significant reduction at p < 0.001 , ** Significant reduction at p < 0.05

Acute Toxicity studies reveals that, other four
extracts were found to have LD50 of solvent ether
1258.92 0.023, ethyl acetate 1258.92 0.023 ,
butanone1778.27 0.017 , n-butanole1778.27
0.204 . Based on these results the LD50 doses of
four extracts were extrapolated to rats, 1/10th of
these were taken as therapeutic doses.
The effect of test extracts of Hedyotis corymbosa
on rats intoxicated with CCL4 are recorded [table
1]. The values are represented as mean SE. The
statistical significance was computed between
CCL4 treated and various extracts treated groups
using unpaired students “t” test. intoxification
with CCL4 results in increase the levels of SGPT
and SGOT , it is evident from the [ table 1 ] . All the
values are elevated in CCL4 treated group
compared with control groups indicating that
CCL4 intoxication resulted in liver damage.
Discussion
The liver can be injured by many chemicals and
drugs. In the present study CCL4 was selected as a
hepatotoxicant to induce liver damage. The
experimental intoxication induced by carbon
tetrachloride (CCl4) is widely used for modeling
liver injury in rats. Hepatotoxicity is connected
with severe impairment of cell protection
mechanisms. The location of liver injury is
defined mainly by the biotransformation of CCl4,
which is cytochrome P-450 dependant and leading
to the generation of an unstable complex CCl3
radical. Free radicals initiate the process of lipid
peroxidation, which is generally cause of
inhibition of enzyme activity[12,13]. Therefore,
SGPT is more specific to the liver, and is thus a
better parameter for detecting liver injury .The
elevated levels of SGOT and SGPT were
significantly decreased by ether and butanol at P <
0.001 and butanone and ethanol at P < 0.05, where
as petroleum ether and ethyl acetate did not shown
any significant reduction in the level of SGOT and
SGPT. Similarly Liv -52 a standard drug also
shown significant reduction of SGOT and SGPT at
P< 0.001. Among the various extracts used in this
study, only petroleum ether and ethyl acetate
extracts has not shown any significant activity.
Where as other extracts like ether and butanol
shown significant hepatoprotective activity
compared to the standard drug.

pages 32 - 35
 ANCIENT SCIENCE OF LIFE
It can be concluded from the enzymic levels and
histopathological observation that solvent ether,
butanol, ethanol, butanone extracts of Hedyotis
corymbosa imparts hepatoprotective activity in
CCL4 induced injury.
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