Karupannan Arumugasamy et al.

IRJP 2011, 2 (10), 19-21

INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(10), 2011
INTERNATIONAL RESEARCH JOURNAL OF PHARMACY ISSN 2230 8407
Available online www.irjponline.com Research Article

EFFECT OF ANISOCHILUS DYSOPHYLLOIDES BENTH. (LAMIACEAE) ON CARBONTETRA
CHLORIDE (CCl
4
) INDUCED HEPATOTOXICITY IN RATS
Pandancode Chandran Sajitha, Karupannan Arumugasamy*, Uthaman Danya, Thimmiyan Baluprakash and
Madathupatti Ramanathan Udhayasankar
Kongunadu arts and science college, Coimbatore, Tamilnadu, India

Article Received on: 11/08/11 Revised on: 25/09/11 Approved for publication: 20/10/11

*Email: arumugasamy_kasc@yahoo.co.in

ABSTRACT
The hepatoprotective effect of Anisochilus dysophylloides was evaluated on carbon tetrachloride (CCl4) induced in rats. The liver marker enzymes such as serum glutamate
oxaloacetate (SGOT), serum glutamate pyruvate transaminase (SGPT) and serum alkaline phosaphate (serum ALP) levels were significantly increased in Group II while the
serum total protein, albumin and globulin levels were significantly decreased in Group II. The SGOT, SGPT, ALP and bilirubin levels were significantly decreased in Group III
and IV at the dose of 100 and 200mg/kg b.wt. However, the serum total protein, albumin and globulin levels were elevated in the same Group II and IV. A. dysophylloides
ethanolic extract also neutralized lipid peroxidation (LPO), glutathione peroxidase (GPX), glutathione reductase GRD, super oxide dismutase (SOD) and catalase (CAT) levels at
a dose of 200mg/kg b.wt. Silymarin used as standard drug also exhibited significant hepatoprotective activity on post treatment against CCl4 hepatotoxicity in rats. The results of
this study strongly indicate that A. dysophylloides has a potent hepatoprotective action against carbontetra chloride induced hepatic damage in rats.
KEYWORDS Anisochilus dysophylloides, carbon tetrachloride, SGOT, SGPT, SALP, Silymarin.

INTRODUCTION
The herbal drugs have been used in the treatment of liver diseases in
China and India. Some herbal drugs promoted several disorders and
contain potent hepatotoxic alkaloids and are harmful
1
. Many
traditional remedies employ herbal drugs for the treatment of liver
ailments
2
. The family Lamiaceae has traditionally occupied an
important position in the socio- cultural, spiritual and medicinal
arena of rural and tribal lives of India. The leaves of Lamiaceae
members are used as diaphoretic, expectorant, ear ache, cure skin
orders
3
, anti-inflammatory, antipyretic
4
, chemoprotective
5
,
antioxidant
6
, analgesic, immunomodulatory
7
, antiarthritic,
hypoglycemic and hypolipidaemic activities
8
. Anisochilus
dysophylloides Benth. (Lamiaceae) has been used in ayurvedic
system of medicine and each part of the plant has medicinal value.
However, so far to our knowledge there are no reports on the effect
of A. dysophylloides extract on hepatoprotective activity. Therefore,
the present study is focused on assessing hepatoprotective activity of
A. dysophylloides ethanolic extract.
MATERIALS AND METHODS
Plant Material
Whole plant of Anisochilus dysophylloides was collected from
Coonor, the Nilgiris district, Coimbatore, Tamilnadu and
authenticated by a taxonomist, Murugasen, Senior scientist,
SACON, Coimbatore. The plant material was air-dried at room
temperature. The dried material was first extracted with petroleum
ether and followed by ethanol. The ethanolic extract was
concentrated at room temperature in amber-coloured light protected
bottles for the further experimental studies.
CCl
4
induced Hepatotoxicity
Male albino rats weighing about (150-200gm) were used in this
study. They were kept in a 12:12hr L: D cycle and the temperature
maintained at 222C. Standard laboratory pellet diet (Hindustan
Lever, Bangalore) and water was administered to the rats ad libitum.
The animals were taken care as per the Guidelines for the care and
use of animals in the scientific research by the Indian National
Science Academy New Delhi
9
.
The rats were randomly divided into 5 groups, each group
containing six animals, as given below:
Group I Control rats
Group II CCl
4
induced liver injured rats (carbontetra chloride
1ml/kg b.wt.)
Group III Liver injured rats administered with A. dysophylloides
drug (100mg/kg b.wt.)
Group IV Liver injured rats administered with A. dysophylloides
drug (200 mg/kg b.wt.)
Group V Rats administered with a standard drug silymarin
(100g/kg b.wt.).
Animals were sacrificed 48h after the last injection. Blood was
collected, allowed to clot and serum separated. It is used for further
biochemical studies.
RESULTS AND DISCUSSION
The present study showed that the liver marker enzymes SGOT,
SGPT and ALP levels were significantly increased in Group II,
while the serum total protein, albumin and globulin levels were
significantly decreased in Group II when compared to control rats.
The ethanolic extract of A. dysophylloides possess hepatoprotective
activity as evidenced by the significant decrease in the liver marker
enzymes SGOT, SGPT, ALP and bilirubin levels in Group III and
IV compared to Group II. However, the serum total protein, albumin
and globulin levels were elevated in the rats administered with 100
and 200mg/kg b.wt. (Table I-III and Figs I-III). In ayurvedic
medicine, the plant has been recommended for the treatment of
hepatic disorder. Earlier studies have shown that several medicinal
plants have hepatoprotective activities
10
.
The LPO levels were significantly increased in Group II CCl
4
induced hepatotoxicity rats, compared to the control group I.
Treatment of liver injured rats with AD extract at the dose of
100mg/kg b.wt. in group III and 200mg/kg b.wt. in group IV showed
a significant increase when compared to the control group. This
result is compared to the liver injured rats received silymarin orally
at the dose of 100mg/kg b.wt. for 7 days. GPx, GRD, SOD and CAT
were significantly decreased in AD treated liver injured group III
(100mg/kg b.wt.) and group IV(200mg/kg b.wt.) as compared to the
control group I. (Table IV; Fig IV).
CCl
4
induced hepatotoxicity is a widely used animal model to assess
the hepatoprotecive activity of drug. The hepatoprotective effect of
CCl
4
are thought to result from its reductive dehalogenation by the
P450 enzyme system to the highly reactive free radical CCl
3.
This
radical combines with molecular oxygen to form the trichloromethyl
peroxyl radical which abstracts hydrogen from unsaturated lipids
and initiates the process of lipid peroxidation
11
. Serum ALP and
bilirubin is also associated with liver cell damage. The ALP activity
Karupannan Arumugasamy et al. IRJP 2011, 2 (10), 19-21
INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(10), 2011
and serum bilirubin level are largely used as most common
biochemical markers to evaluate liver injury
12
. The administration of
A. dysophylloides extracts has prevented the increased serum marker
enzyme ALP and bilirubin level. This is in agreement with the
commonly accepted view that serum levels of ALP return to normal
with the healing of hepatic parenchyma and the regeneration of
hepatocysts
13
. An increase in LPO indicates serious damage to cell
membranes, inhibition of several enzymes and cellular function
14
. In
the present study, an increase of LPO level was found in the CCl
4

induced rats. It is well known that SOD, CAT, GPx and GRD play
an important role as protective enzyme against free radical formation
in tissues
15
.
The hepatoprotective activity of Anisochilus dysophylloides
ethanolic extract is effective against CCl
4
induced hepatic damage.
This herbal drug alleviates the symptoms of liver damage, as evident
by biochemical assay and seems to raise some concern about the
traditional indications of this species as a medicine for liver disease.
However, further studies need to be worked out with other
hepatotoxic compounds and evaluating other toxicity biomarkers.
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Table I Effect of AD extracts on the total protein, albumin, globulin
concentration in the normal, liver damaged and drug treated rats
GROUPS Total Protein (mg/dl) Albumin (g/dl) Globulin (g/dl)
GROUP I 8.34 0.34 4.530.31 3.810.34
GROUP II 5.11 0.14* 3.900.14 1.210.10*
GROUP III 5.91 0.21 3.100.13 2.810.45
GROUP IV 6.13 0.4 3.130.11 3.000.32
GROUP V 7.99 0.15a 4.510.21 3.480.33a
Group I Control rats given normal saline daily for 21 days
Group II CCL4 induced liver toxicity in rats given normal saline daily for 21days
Group III Liver injured rats given AD drug at the dose of 100 mg/ Kg body weight
daily orally for 21 days
Group IV Liver injured rats given AD drug at the dose of 200 mg/ Kg body weight
daily orally for 21 days
Group V Liver injured rats received silymarin (100 g/kg b wt / day) orally.

Table II Effect of AD extracts on enzyme activity of serum GOT, GPT and ALP
in the normal, liver damaged and drug treated rats
GROUPS SGOT
(U/L)
SGPT
(U/L)
ALP
(U/L)
GROUP I 32.141.71 28.141.31 136.514.64
GROUP II 138.533.16* 93.511.64* 198.633.16*
GROUP III 121.612.84* 70.151.36* 163.263.56
GROUP IV 62.511.16a 43.211.14a 131.564.11a
GROUP V 30.251.91a 21.561.31a 114.313.56a
Group I Control rats given normal saline daily for 21 days
Group II CCL4 induced liver toxicity in rats given normal saline daily for 21days
Group III Liver injured rats given AD drug at the dose of 100 mg/ Kg body weight
daily orally for 21 days
Group IV Liver injured rats given AD drug at the dose of 200 mg/ Kg body weight
daily orally for 21 days
Group V Liver injured rats received silymarin (100 g/kg b wt / day) orally.

Table III Effect of AD extracts on the serum Total, conjugated and
unconjugated bilirubin levels in the normal control, liver injured and drug
treated rats.
GROUPS Total Bilirubin
(mol/L)
Conjugated
(mol/L)
Unconjugated
(mol/L)
GROUP I 0.840.14 0.240.11 0.600.15
GROUP II 2.630.21 1.030.16 1.600.18
GROUP III 1.830.14 0.800.11 1.030.17
GROUP IV 1.050.24 0.250.13 0.800.10
GROUP V 0.810.05 0.210.11 0.600.16
Group I Control rats given normal saline daily for 21 days
Group II CCL4 induced liver toxicity in rats given normal saline daily for 21days
Group III Liver injured rats given AD drug at the dose of 100 mg/ Kg body weight
daily orally for 21 days
Group IV Liver injured rats given AD drug at the dose of 200 mg/ Kg body weight
daily orally for 21 days
Group V Liver injured rats received silymarin (100 g/kg b wt / day) orally.

Table IV Effect of AD extracts on liver LPO, GPX, GRD, SOD and CAT in the normal control, liver injured and drug treated rats.

GROUPS
LPO (n mole of MDA/mg
protein)
GPX (/mg
Protein)
GRD (/mg) SOD ( /mg)

CAT (/mg)

GROUP I 0.890.03 8.510.13 8.110.03 6.840.03 10.511.05
GROUP II 3.940.04* 3.050.21** 2.630.02* 1.940.01** 4.110.98**
GROUP III 3.050.06* 3.920.14* 3.610.01* 3.560.05 5.690.91*
GROUP IV 1.930.05 5.690.11a 5.110.04a 4.910.03a 8.650.84a
GROUP V 0.910.03aa 8.940.15aa 9.260.02aa 7.530.02a 9.980.15a
Group I-Control rats given normal saline daily for 21 days
Group II-CCL4 induced liver toxicity in rats given normal saline daily for 21days
Group III-Liver injured rats given AD drug at the dose of 100 mg/ Kg body weight daily orally for 21 days
Group IV- Liver injured rats given AD drug at the dose of 200 mg/ Kg body weight daily orally for 21 days
Group V-Liver injured rats received silymarin (100 g/kg b wt / day) orally.

Karupannan Arumugasamy et al. IRJP 2011, 2 (10), 19-21
INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(10), 2011


Fig I Effect of AD extracts on the protein, albumin, globulin concentration in the normal, liver damaged and drug treated rats.



Fig II Effect of AD extracts on enzyme activity of serum GOT, GPT and ALP in the normal, liver damaged and drug treated rats.



Fig III Effect of AD extracts on the serum Total, conjugated and unconjugated bilirubin levels in the normal control, liver injured and drug treated rats.



Fig IV Effect of AD extracts on liver LPO, GPX, GRD, SOD and CAT in the normal control, liver injured and drug treated rats.

Source of support: Nil, Conflict of interest: None Declared