See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/268268248

Protective effect of the roots of Capparis sepiaria Linn. (Himsra) on carbon

tetrachloride-induced hepatotoxicity

Article

CITATION READS

1 150

4 authors, including:

Anita Murali Sunkam Yoganarasimhan

Faculty of Pharmacy, M. S. Ramaiah University of Applied Sciences M.S. Ramaiah College of Pharmacy
26 PUBLICATIONS   132 CITATIONS    70 PUBLICATIONS   332 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Pharmacognosy of novel potential Medicinal Plants View project

All content following this page was uploaded by Anita Murali on 28 March 2015.

The user has requested enhancement of the downloaded file.

 Hepatoprotective activity of Capparis sepiaria / Asian Journal of Traditional Medicines, 2012, 7(1)

Regular articles

Protective effect of the roots of Capparis sepiaria Linn.

(Himsra) on carbon tetrachloride-induced hepatotoxicity
Varadharajan Madhavan a, Anita Murali b*, Sunkam Yoganarsimhan a, Ajay Shankar Pandey a
a. Department of Pharmacognosy, M S Ramaiah College of Pharmacy, Bangalore 560054, India
b. Department of Pharmacology, M S Ramaiah College of Pharmacy, Bangalore 560054, India

Abstract
Capparis sepiaria L. is an important drug used in Ayurvedic medicine. In the present study extracts of the roots of C. sepiaria
were evaluated for their hepatoprotective potential on carbon tetrachloride (CCl4)-induced hepatotoxicity in albino Wistar
rats. The extent of hepatoprotection was evaluated by estimating the serum levels of hepatic transaminases (SGPT and SGOT),
alkaline phosphatase (ALP), total protein (TP), and bilirubin (total and direct). Aqueous and ethanol extracts significantly
reduced the increased liver weight as well as the serum levels of SGPT, SGOT, ALP, and bilirubin and normalized the reduced
serum protein levels in the treated rats as supported by histopathological studies. The extracts were also subjected to preliminary
organic analysis and chromatographic studies including an HPTLC finger print profile. The results obtained indicate that the
roots of C. sepiaria have a significant hepatoprotective effect on CCl4-induced hepatotoxicity.

Key words: Capparis sepiaria; carbontetrachloride induced hepatotoxicity; histopathology; roots; HPTLC

1. Introduction hepatic changes induced by CCl 4 resemble those

of viral hepatitis [3]. CCl 4 is biotransformed by
The liver is a vital organ involved in the cytochrome P450 enzymes in the endoplasmic
maintenance of homeostasis in the body and in reticulum of liver cells to a highly reactive
various biochemical pathways [1]. Liver injury is trichloromethyl free radical which in turn reacts with
caused when certain medicinal agents are consumed oxygen to form a trichloromethyl peroxy radical [4].
in overdose or even within the therapeutic range.
Chemicals that cause liver injury are called Himsra is an important drug used in several
hepatotoxins and more than 900 drugs have been formulations in Ayurvedic medicine and its botanical
implicated in causing liver injury [2]. Carbon source is Capparis sepiaria L. (Capparaceae) [5].
tetrachloride is commonly used for the induction Himsra possesses properties like ‘Shothhar’ (anti-
of hepatotoxocity in experimental animals and the inflammatory), ‘Vedanasthapan’ (analgesic),
‘Raktashodhak’ (blood purifier), ‘Hrudayottejak’
(cardiotonic) and is used in the management of
* Author to whom correspondence should be addressed. Address: ‘Yakrat dosha’ (liver disorders), ‘Pachanvikar’
Department of Pharmacology, M. S. Ramaiah College of Pharmacy,
(digestive disorders), diseases caused by ‘Kapha-
MSRIT Post, Bangalore 560054, Karnataka, India; Tel.:
+91-80-23608942, 9845105865; Fax: +91-80-23607488; Email: Vata’ imbalance, Shlipad (filariasis), cardiac
[email protected]
weakness, fever and to reduce swellings of the
Received: 2011-06-24 Accepted: 2012-01-01 testicles. The seeds are used as an antidote to snake

18
 Hepatoprotective activity of Capparis sepiaria / Asian Journal of Traditional Medicines, 2012, 7(1)

bite, roots in the treatment of mumps and root bark sexes were used for the study. Inbred animals were
in scabies and eczema [6]. reared and housed in the animal house of MSRCP
and were well maintained under standard conditions
The leaves of C. sepiaria are reported to contain of temperature (22 ± 2ºC), room humidity (60 ±
four pentacyclic triterpenoid alcohols, taraxasterol, 10%) and a 12 h day and night cycle, and given
α and β-amyrin, β-sitosterol, erythrodiol [7] and food (rat pellets from VRK Nutritional Solutions,
betulin [8] and a complex alkaloid has been reported Sangli, Maharashtra, India) and water ad libitum.
in the roots [9]. The anti-inflammatory and analgesic The study protocol was approved by the Institutional
activities of the root, the hepatoprotective activity of Animal Ethical Committee of MSRCP (MSRCP/
the stem, and the antidiabetic activity of the leaves M-34/2009).
have already been reported [10, 11].
2.3. Chemicals and reagents
Himsra (C. sepiaria) is used for the treatment of
liver troubles in ayurvedic medicines. The present Serum parameters were estimated using analysis
work on the hepatoprotective property of the roots is kits purchased from Span diagnostics Ltd, Surat,
undertaken to confirm this activity. India (SGOT, SGPR and TP) and ERBA Diagnostics
Mannheim GmbH, Germany (ALP and bilirubin).
2. Materials and methods Silymarin was a gift from Micro Labs, Hosur, Tamil
Nadu, India. CCl4 and liquid paraffin were obtained
2.1 Plant material from Merck Ltd, Mumbai; anaesthetic ether was
from TKM Pharma, Hyderabad, India; and ethanol
The roots of C. sepiaria were collected from and chloroform were from Rankem Ltd, New Delhi,
the forests of Aralvoymozhi, Kanyakumari district India.
of Tamil Nadu, in April 2010, and identified
and authenticated by Dr. S. N. Yoganarasimhan,
2.4. Preparation of extracts
Taxonomist and Research co-ordinator, M S Ramaiah 2.4.1. Aqueous extract
College of Pharmacy (MSRCP), Bangalore, India.
The coarsely powdered root (200 g) of
The voucher herbarium specimen (Ajay Shankar
C. sepiaria was extracted by maceration with
Pandey 035) has been deposited in the herbarium of
chloroform-water (0.25% v/v of chloroform
the Department of Pharmacognosy, MSRCP, along
in distilled water) followed by filtration and
with a sample of the crude drug in the crude drug
concentration of the extract to dryness to obtain a
museum. The taxonomic identification was carried
dark brown semisolid mass (yield 10.3% w/w). The
out using local flora [12, 13].
aqueous extract for pharmacological studies was
2.2. Animals prepared using a 2% w/v acacia solution in distilled
water.
The pharmacological experiments were carried
out following the guidelines specified by the 2.4.2 Ethanol extract
Committee for the Purpose of Co-ordination and
The coarsely powdered roots (200 g) of C.
Supervision of Experiments on Animals (CPCSEA),
sepiaria were extracted with 95% v/v ethanol in a
Ministry of Environment and Forests (Animal
soxhlet apparatus by continuous hot extraction. The
Welfare Division), Govt of India, New Delhi. Albino
extract was filtered and concentrated to dryness
rats (Wistar strain) weighing 170-250 g of both

19
 Hepatoprotective activity of Capparis sepiaria / Asian Journal of Traditional Medicines, 2012, 7(1)

under reduced pressure to obtain a dark brown follows:

semisolid mass (yield 9.4% w/w). The dried extract
was made into a suspension in 2% w/v acacia using Group 1: Vehicle control (Distilled water
distilled water. containing 2% w/v acacia) p.o.

The aqueous and ethanol extracts were Group 2: Positive control (CCl4 1.5 ml/kg body
subjected to preliminary organic analysis [13] and weight, p.o. as a single dose).
chromatographic studies [14]. The presence of Group 3: Standard - silymarin (100 mg/kg body
flavonoids was confirmed by HPTLC fingerprint weight p.o.) + CCl4 (1.5 ml/kg body weight, p.o.) as
studies. a single dose.
2.5. HPTLC fingerprint studies
Group 4: Aqueous extract (200 mg/kg body
HPTLC studies were carried out using a weight, p.o.) + CCl4 (1.5 ml/kg body weight, p.o.) as
Camag HPTLC system equipped with a Linomat V a single dose.
applicator, TLC scanner 3 and Reprostar 3 with a 12
Group 5: Aqueous extract (400 mg/kg body
bit CCD camera for photographic documentation,
weight, p.o.) + CCl4 (1.5 ml/kg body weight, p.o.) as
using WINCATS-4 software. All the solvents used
a single dose.
were of HPLC grade and obtained from MERCK.
Ethanol and aqueous extracts (4 µl) were applied Group 6: Ethanol extract (200 mg/kg body
to precoated silica-gel F 254 plates (Merck) as 8 weight, p.o.) + CCl4 (1.5 ml/kg body weight, p.o.) as
mm bands using a Linomat V applicator with a a single dose.
hamilton syringe and developed in a Camag HPTLC
twin trough linear development chamber using Group 7: Ethanol extract (400 mg/kg body
toluene: dioxane: glacial acetic acid (90:25:4). The weight, p.o.) + CCl4 (1.5 ml/kg body weight, p.o.) as
plates were scanned at 254 and 366 nm and photo- a single dose.
documented using a Camag Reprostar 3 [15].
The control animals received acacia solution
2.6. Acute toxicity studies (2% w/v) in distilled water. Group 3 animals were
given silymarin (100 mg/kg body weight p.o.) once
Acute toxicity studies were performed on
daily for 7 days. Group 4 to 7 were given aqueous
aqueous and ethanol extracts following OECD
extract (200 and 400 mg/kg) and ethanol extract (200
guidelines (423) [16]. The highest dose tested
and 400 mg/kg), respectively, once daily for 7 days.
was 2000 mg/kg for both extracts. Accordingly,
All the groups except the normal control were given
1/10th and 1/5th of this dose were selected for the
CCl4 (1.5 ml/kg, p.o.) once on the 7th day to induce
pharmacological studies.
acute liver damage. On the 8th day, 18 h after the last
2.7. CCl4 - induced hepatotoxicity dose of CCl4, all the animals were put under light
ether anaesthesia and blood was collected from the
2.7.1. Experimental protocol
retro-orbital sinus using a heparinized capillary tube.
Healthy Albino Wistar rats of both sexes (7-8 The blood was allowed to clot and the separated
weeks old) weighing 170-250 g were used. They serum was obtained by centrifugation at 12000 rpm
were randomized into 7 groups of 6 animals each as for 10 min. The separated serum was used for the

20
 Hepatoprotective activity of Capparis sepiaria / Asian Journal of Traditional Medicines, 2012, 7(1)

estimation of total bilirubin, direct bilirubin, SGOT, The ethanol extract contained 3 phytoconstituents
SGPT, ALP and total proteins (TP). The animals having Rf values of 0.63, 0.83 and 0.94 under 254 nm
were sacrificed by administering an excess of ether and 11 phytoconstituents having Rf values of 0.37,
and their livers were removed. 0.39, 0.47, 0.58, 0.69, 0.75, 0.80, 0.83, 0.88, 0.92,
and 0.96 under 366 nm (Figs. 1 & 2).
2.7.2. Isolation of liver

Liver was carefully removed and washed with The aqueous extract contained 6
ice cold saline solution and pressed between pads phytoconstituents having Rf values of 0.02, 0.22,
of filter paper and weighed. A portion of the liver 0.47, 0.63, 0.81, and 0.94 under 254 nm and 12
was preserved in 10% v/v neutral formalin for phytoconstituents having Rf values of 0.02, 0.13,
histopathological studies. 0.18, 0.22, 0.29, 0.38, 0.60, 0.69, 0.79, 0.82, 0.89,
and 0.96 under 366 nm (Figs. 3 & 4).
2.7.3. Biochemical estimations
3.2. Acute toxicity studies
Biochemical estimations were performed for
None of the animals treated with either of the
ALT [17], AST [18], ALP [19], TP [20, 21] and
extracts died over a period of one week and until the
bilirubin [22, 23] using a Semi-automatic Chemistry
end of the study.
Analyzer (CA-2005, B4B® Diagnostic Division),
Ranbaxy Diagnostic Division, India.

2.7.4. Histopathological studies

Paraffin sections were prepared from formalin-

fixed liver samples and stained with haematoxylin
and eosin. Histopathology category scores were
AU

assigned based on the extent of injury.

2.8. Statistical analysis

The data were expressed as mean±SEM values
and tested with one way analysis of variance Rf

(ANOVA) followed by the Tukey-Kramer multiple Peak Start Start Max Max Max
comparison test. Position Height Position Height (%)
(Rff) (AU) (Rf) (AU)
1 0.60 9.9 0.63 19.7 13.19
3. Results 2 0.80 23.3 0.83 40.4 27.12
3 0.90 39.6 0.94 88.9 59.69
3.1. Phytochemical and chromatographic studies Peak End End Area Area
Position Height (AU) (%)
Preliminary phytochemical studies revealed the (Rf) (AU)
presence of alkaloids, carbohydrates, phytosterols, 1 0.67 13.6 872.7 16.04
2 0.85 33.3 1468.0 26.99
saponins, flavonoids, gums and mucilage (Table 1).
3 0.97 21.0 3099.0 56.97
HPTLC fingerprint profiles for flavonoids in aqueous
and ethanol extracts of C. sepiaria were obtained Fig. 1. HPTLC fingerprint of the alcohol extract of C.
and Rf values recorded. sepiaria at 254 nm

21
 Hepatoprotective activity of Capparis sepiaria / Asian Journal of Traditional Medicines, 2012, 7(1)

3.3. CCl4- induced hepatotoxicity and direct bilirubin were estimated and compared
with those of the positive controls. The liver weight
The serum levels of SGPT, SGOT, ALP, TP, total
and levels of SGPT, SGOT, ALP and bilirubin
significantly increased (P<0.05 for liver weight and
P<0.001 for the other parameters) while the level of
total proteins was significantly (P<0.001) reduced
in the positive control group. Treatment with drug
extracts produced significant changes in the altered
AU

serum parameters, confiming the hepatoprotective

activity of the drug. Elevated SGPT levels were
significantly reduced in the extract-treated groups
(P<0.01 for 200 mg/kg and P<0.001 for 400 mg/kg).
SGOT levels were also significantly reduced in the
Rf

Peak Start Start Max Max Max

Position Height Position Height (%)
(Rf) (AU) (Rf) (AU)
1 0.36 1.2 0.37 27.9 4.24
2 0.38 1.4 0.39 15.2 2.30
3 0.46 2.3 0.47 148.5 22.56
AU

4 0.56 4.9 0.58 68.5 10.41

5 0.67 7.8 0.69 79.9 12.14
6 0.72 8.4 0.75 69.8 10.61
7 0.79 6.4 0.80 105.0 15.95
8 0.83 9.5 0.83 24.5 3.72 Rf
9 0.84 10.4 0.88 47.2 7.17
Peak Start Start Max Max Max
10 0.89 18.8 0.92 31.2 4.75 Position Height Position Height (%)
11 0.93 31.2 0.96 40.5 6.15 (Rf) (AU) (Rff) (AU)
1 0.01 0.05 0.02 57.0 15.13
Peak End End Area Area
2 0.21 2.7 0.22 12.0 3.19
Position Height (AU) (%)
(Rf) (AU) 3 0.47 0.7 0.47 14.0 3.72
1 0.37 1.1 145.6 2.04 4 0.60 11.9 0.63 23.3 6.18
2 0.40 1.7 84.8 1.19 5 0.76 12.1 0.81 153.4 40.70
3 0.49 2.2 808.8 11.34 6 0.90 39.2 0.94 117.2 31.08
4 0.59 6.4 506.2 7.10 Peak End End Area Area
5 0.70 7.7 606.1 8.50 Position Height (AU) (%)
(Rf) (AU)
6 0.78 7.4 1028.4 14.41
1 0.05 1.9 600.0 5.72
7 0.81 8.0 574.2 8.05
2 0.25 0.1 121.4 1.16
8 0.84 10.2 203.0 2.85
3 0.48 1.1 86.5 0.83
9 0.89 18.4 671.4 9.41 4 0.65 15.7 698.1 6.66
10 0.93 30.8 930.1 13.04 5 0.85 35.7 5681.5 54.02
11 1.00 5.3 1576.0 22.09 6 0.97 17.0 3313.1 31.61
Fig. 2. HPTLC fingerprint of the alcohol extract of C. Fig. 3. HPTLC fingerprint of the aqueous extract of C.
sepiaria at 366 nm sepiaria at 254 nm

22
 Hepatoprotective activity of Capparis sepiaria / Asian Journal of Traditional Medicines, 2012, 7(1)

Table 1. Preliminary phytochemical analysis of C. sepiaria

Phytoconstituents Alcohol extract Aqueous extract

Alkaloids ++ ++
Carbohydrates ++ ++
Phytosterols ++ --
Fixed oils and fats -- --
Phenolic compounds and tannins -- --
Saponins -- ++
Flavonoids ++ ++
Proteins and amino acids -- --
Gums and mucilages -- ++
Note: ++ = Present; -- = Absent

treated groups (P<0.05 and P<0.01, respectively, for and 400 mg/kg P<0.01 and P<0.001, respectively)
200 and 400 mg/kg of the aqueous extract and P<0.01 and in ethanol extract-treated group (200 and 400
and P<0.001, respectively for 200 and 400 mg/kg of mg/kg P<0.001). Serum levels of total protein were
the ethanol extract). ALP levels were significantly reduced in the positive control group. The aqueous
reduced in the aqueous extract- treated groups (200 extract 400 mg/kg (P<0.05), and the ethanol extract

Table 2. Effect of C. sepiaria root extracts on serum biochemical parameters

Groups Liver wt g/ 100 g bw SGPT (U/l) SGOT (U/l) ALP (U/l)

Normal control 2.74±0.12 40.42±1.88 105.93±4.97 151.31±3.12

Positive control (CCl4 1.5 ml/kg) 3.18±0.09 122.18±8.86 333.65±15.64 336.88±24.33
Standard (Silymarin, 100 mg/kg) 2.67±0.48 ns 56.55±0.70*** 132.00±8.04*** 171.19±6.5***
Aq. Extract (200 mg/kg) 3.02±0.15 ns 94.93±2.01** 248.87±27.93* 273.69±6.98**
Aq. Extract (400 mg/kg) 2.78±0.09 ns 63.37±2.58*** 217.53±19.31** 247.78±6.47***
Alc. Extract (200 mg/kg) 2.74±0.11 ns 97.51±5.09** 236.67±21.65** 257.30±2.43***
Alc. Extract (400 mg/kg) 2.89±0.12 ns 65.32±2.15*** 208.43±8.18*** 240.43±4.42***
Bilirubin (mg/dl)
Groups Total protein (g/dl)
Total Direct
Normal control 6.06±0.55 0.43±0.04 0.17±0.02
Positive control (CCl4 1.5 ml/kg) 3.08±0.06 2.31±0.18 0.67±0.06
Standard (Silymarin, 100 mg/kg) 5.52±0.19*** 0.71±0.06*** 0.21±0.01***
Aq. Extract (200 mg/kg) 3.83±0.27 ns 1.85±0.05** 0.53±0.02*
Aq. Extract (400 mg/kg) 4.58±0.28* 1.79±0.04** 0.47±0.03**
Alc. Extract (200 mg/kg) 4.54±0.22* 1.76±0.05** 0.48±0.02**
Alc. Extract (400 mg/kg) 4.98±0.29** 1.61±0.09*** 0.36±0.02***
Values are expressed as Mean ± SEM, Data is compared against positive control group. One way analysis of variance (ANOVA) P value
for liver weight per 100 g of body weight considered non significant with 0.4247; P value for SGPT, SGOT, alkaline phosphatase, total
protein, total bilirubin and direct bilirubin considered extremely significant with P<0.0001. Tukey-Kramer multiple comparison test. ***
P< 0.001, ** P< 0.01, * P< 0.05; Note: U/l = Units per litre; ns = non-significant.

23
 Hepatoprotective activity of Capparis sepiaria / Asian Journal of Traditional Medicines, 2012, 7(1)

the positive control group and there was a significant

decrease in total bilirubin levels in the aqueous
extract-treated groups, 200 and 400 mg/kg (P<0.01),
AU

and the ethanol extract-treated groups, 200 and 400

mg/kg (P<0.01 and P<0.001, respectively). Direct
bilirubin levels were significantly reduced in the
aqueous extract-treated groups, 200 and 400 mg/kg
Rf
(P<0.05, P<0.01, respectively) and in the ethanol
Start Start Max Max Max
Peak Position Height Position Height (%) extract-treated groups, 200 and 400 mg/kg (P<0.01,
(Rf) (AU) (Rf) (AU) P<0.001, respectively). The liver weight also
1 0.01 1.6 0.02 75.3 12.36
decreased following treatment with the drug extracts
2 0.12 0.5 0.13 70.2 11.52
(Table 2).
3 0.17 0.7 0.18 56.4 9.25
4 0.20 0.7 0.22 18.9 3.10 3.3.1. Histopathological studies
5 0.27 0.1 0.29 103.8 17.04
6 0.38 0.8 0.38 17.5 2.87 Histopathological observations clearly revealed
7 0.58 5.4 0.60 16.4 2.69 the extent of hepatic damage caused by CCl 4
8 0.68 7.8 0.69 72.6 11.92 and also the extent of the hepatoprotective effect
9 0.78 4.3 0.79 20.8 3.42 in the form of reparative changes. The liver of
10 0.81 7.1 0.82 52.8 8.66 normal control animals exhibited normal hepatic
11 0.84 6.5 0.89 66.8 10.96 architecture with the central veins, portal tracts,
12 0.91 27.6 0.96 37.7 6.19 hepatocytes and sinusoids appearing normal. The
End End Area Area lobular unit was clearly identified. The positive
Peak Position Height (AU) (%)
(Rf) (AU) control group showed a loss of normal liver
1 0.04 3.0 941.1 12.97 architecture with degenerative hepatocytes and
2 0.15 0.1 461.3 6.36 moderate fibrosis and the sinusoidal spaces were
3 0.19 0.2 283.9 3.91 flooded with inflammatory cells. The liver of
4 0.25 0.3 165.7 2.28 aqueous extract-treated group (200 mg/kg) showed
5 0.31 1.3 1029.6 14.19
moderately diffused degeneration of hepatocytes
6 0.40 3.7 104.5 1.44
and mild to moderate fibrosis. The aqueous extract-
7 0.60 7.2 161.9 2.23
treated group (400 mg/kg) showed mild fibrosis
8 0.71 7.5 709.07 9.77
and hepatocyte degeneration. An improved liver
9 0.80 7.1 204.4 2.82
histology as found in the ethanol extract-treated
10 0.83 6.4 358.9 4.95
groups. Photomicrograph images of the ethanol
11 0.91 26.8 1036.4 14.29
12 1.00 1.2 1797.6 24.78
extract-treated group (200 mg/kg) showed only
mild fibrosis and moderately diffused degeneration
Fig. 4. HPTLC fingerprint of the aqueous extract of C. sepiaria
of hepatocytes. The rats treated with the ethanol
at 366 nm
extract (400 mg/kg) showed minimal fibrosis and
focal degeneration of hepatocytes. The standard
200 and 400 mg/kg (P<0.05and P<0.0.1respectively)
silymarin-treated group showed a reversal of toxicity
increased the altered protein level significantly.
with minimal fibrosis and a lesser degree of focal
Serum bilirubin levels (total and direct) increased in

24
 Hepatoprotective activity of Capparis sepiaria / Asian Journal of Traditional Medicines, 2012, 7(1)

Fig. 5-11: Histopathological studies of the liver

degeneration of hepatocytes (Figs. 5 to11). is the commonest form of iatrogenic disease. Liver
injury due to CCl4 in rats was first reported in 1936
4. Discussion [24] and this has been widely and successfully
used by many investigators [25, 26]. Carbon
Hepatotoxicity caused by drugs and chemicals tetrachloride is metabolized by cytochrome P-450

25
 Hepatoprotective activity of Capparis sepiaria / Asian Journal of Traditional Medicines, 2012, 7(1)

in the endoplasmic reticulum and mitochondria properties. Hence, these phytoconstituents could
with the formation of CCl3O- , a reactive oxidative have contributed to the hepatoprotective effect.
free radical, which initiates lipid peroxidation Further investigation is needed to identify the
[27, 28]. Administration of a single dose of CCl4 type of phytoconstituent(s) responsible for the
produces a centrilobular necrosis and fatty changes hepatoprotective activity and to identify the exact
in the liver within 24 h [29]. The toxin reaches mechanism of action.
its maximum concentration in the liver within 3
h of administration. Subsequently, the level falls 5. Conclusion
and by 24 h, no CCl4 is left in the liver [30]. CCl4
administration causes an increase in liver weight Both ethanol and aqueous extracts of the roots
and volume [31]. When the cell membrane of of C. sepiaria exhibited significant hepatoprotective
hepatocytes is damaged, a variety of enzymes, activity. The ethanol extract produced better
such as SGOT, SGPT, ALP normally located in the protection from hepatotoxicity, which may be due
cytosol, are released into the blood [32]. The use of to the presence of phytosterols. Histopathological
serum transaminase activity as an index of hepatic studies confirmed these findings. This study
damage is widely been used. Liver cells synthesize provided evidence to support the use of the roots of
albumin, fibrinogen, prothrombin, alpha-1- C.sepiaria as a potential drug for the treatment of
antitrypsin, haptoglobin, ceruloplasmin, transferrin, hepatotoxicity.
alpha fetoproteins and acute phase reactant proteins.
The blood levels of these plasma proteins are Acknowledgements
reduced following extensive liver damage. The
reduction in the protein levels is likely due to The authors thank the Gokula Education
the impairment of protein synthesis during liver Foundation for their support and help provided in
toxicity [33]. Estimation of bilirubin, a metabolic carrying out this work. The authors also thank V.
product of the breakdown of heme, is an important Chelladurai for supplying authentic plant material
parameter in liver function tests. The bilirubin level and Micro labs Ltd for the gift of silymarin.
rises in diseases of hepatocytes, obstruction of
biliary excretion into the duodenum, in hemolysis References
and defects of hepatic uptake and conjugation of [1] Stine KE, Brown TM. The Principles of Toxicology. 2nd
bilirubin , such as Gilbert’s disease [34]. The findings ed. London: CRC Lewis Publications, 1996, 149-159.
[2] Friedman, Scott E, Grendell, James H, McQuaid,
of the present study indicate that both ethanol and
Kenneth R. Current diagnosis & treatment in
aqueous extracts of the roots of C. sepiaria exhibit gastroenterology; New York: Lang Medical Books/
significant hepatoprotective activity which was McGraw-Hill, 2003, 664–679.
further confirmed by histopathological studies. [3] Usha S, Vilasrao S, Kadam J, Ghosh R. Hepatoprotective
Activity of Livobond- A Polyherbal Formulation Against
The phytochemical studies of C. sepiaria showed CCl4 Induced Hepatotoxicity in Rats. Int J Pharmacol,
2008, 4(6): 472-476.
the presence of many important phytoconstituents
[4] Clawson GA. Mechanisms of carbon tetrachloride
like alkaloids, flavonoids, carbohydrates, saponins hepatotoxicity. Immunopathol Res, 1989, 8(2): 104-12.
and phytosterols. Previous studies have reported [5] Sharma PV. Dravyagunavigyan. Vol 2. Varanasi:
that alkaloids [35], flavonoids [36] and saponins Chaukhambha Bharati Academy, 2005, 230-231.
[6] Aditya Chaudhury N, Ghosh D. Studies on insecticidal
[37] are responsible for the hepatoprotective
plants: chemical examination of the leaves of Capparis

26
 Hepatoprotective activity of Capparis sepiaria / Asian Journal of Traditional Medicines, 2012, 7(1)

sepiaria. J Indian Chem Soc, 1970, 47(8): 751-754. A candidate reference method for determination of total
[7] Saraswathy A, Sasikala E, Patra A, Kundu AB. Betulin- protein in serum l. Development and validation. Clin
28-acetate from Capparis sepiaria L. J Indian Chem Chem, 1981, 27: 1642-50.
Soc, 1991, 68(11): 633-34. [22] Pearlman FC, Lee RT. Detection and Measurement of
[8] Chaudhari SR, Chavan MJ, Gaud RS. Phytochemical Total Bilirubin in Serum, with Use of Surfactants as
and pharmacological studies on the roots of Capparis Solubilizing Agents. Clin Chem, 1974, 20(4): 447-453.
sepiaria. Indian J Pharm Sci, 2004, 4(66): 454-457. [23] Henry RJ (Ed). Cinical chemistry: Principles and
[9] Satyanarayan T, Devi K, Mathews AA. Hepatoprotective techniques. 2nd ed. New York: Harper and Row, 1974,
activity of Capparis sepiaria stem against carbon 1042.
tetrachloride-induced hepatotoxicity in rats. J Pharm [24] Nanji AA, Jokelainen K, Foutouhinia M, Rahemtulla A,
Res & HC, 2009, 1(1): 34-35. Thomas P, Tipoe GL. Increased severity of alcoholic
[10] Selvamani P, Latha S, Elayaraja K, Suresh Babu P, liver injury in female rats: role of oxidative stress,
Gupta JK, Pal T K. Antidiabetic activity of the ethanol endotoxin and chemokines. Am J Physiol Gastrointest
extract of Capparis sepiaria L leaves. Indian J Pharm Liver Physiol. 2002, 281(6):G1348- 56.
Sci, 2008, 70(3): 378-380. [25] Cameron GR, Thomas JC and Karunarathe WAE. The
[11] Gamble JS. The Flora of the Presidency of Madras. pathogenesis of liver injury in carbon tetrachloride and
Vol. 1. Dehra Dun: Bishen Singh Mahendra Pal Singh, thioacetamide poisioning. J Path Bact, 1936, 41: 297.
(reprinted), 2005, 46. [26] Handa SS, Sharma A. Hepatoprotective activity of
[12] Nair NC, Henry AH. Flora of Tamil Nadu, India, Series andrographolide from Andrographis paniculata
I: Analysis. Vol I. Coimbatore: Botanical Survey of against carbontetrachloride. Indian J Med Res, 1990,
India, 1983, 14. 92B:276-92.
[13] Harborne JB. Phytochemical Methods. 3rd ed. London: [27] Shirwaiker A, Sreenivasan KK, Krishnanand BR, Kumar
Chapman and Hall, 1998, 4-7. AV. Chemical investigation and anti-hepatotoxic activity
[14] Stahl E. Thin Layer Chromatography. A Laboratory of the root bark of Capparis spinosa. Fitoterapia, 1996,
Hand Book. 2nd ed. Berlin: Springer, 2005, 60-67, 67(3): 200-204.
241-247. [28] Zimmerman MD, Hayman J. Function and integrity
[15] Wagner H, Bladt S. Plant Drug Analysis. 2nd ed. Berlin: of the liver, In: Clinical diagnosis and management
Springer, 1996, 54, 254. by laboratory methods In: Clinical diagnosis and
[16] w w w. i c c v a m . n i e h s . n i h . g o v / S u p p D o c s / O E C D / management by laboratory methods, 17th ed.1976,
OCDE_GL423. Organisation for Economic Cooperation 217-250.
and Development (OECD) Guidelines for Testing of [29] Agarwal AK and Mehendale JK. Potentiation of carbon
Chemicals-423, Acute Oral Toxicity-Acute Toxic Class tetrachloride hepatotoxicity and lethality by chlordecone
Method; 17 Dec, 2001. in female rats. Toxicology, 1983, 26: 231-242.
[17] Schumann G, Bonora R, Ceriotti F, Ferard G, Ferrero [30] Dawkins MJR. Carbon tetrachloride poisoning in
CA, Franck PF, et al. IFCC primary reference procedures the liver of the new-born rat. J Path Bact, 1963, 85:
for the measurement of catalytic activity concentrations 189-196.
of enzymes at 37 °C. Part 4. Reference procedure for [31] Reddy DG, Reddy SKG, Reddy SN, Reddy VR.
the measurement of catalytic concentration of alanine Hepatoprotective activity of Rhus mysorensis against
aminotransferase. Clin Chem Lab Med, 2002, 40: carbon tetrachloride induced hepatotoxicity in albino
718-733. rats. Int J Pharm Sc Rev Res, 2010, 4(3): 46-48.
[18] Bergmeyer HU, Bowers GN, Horder M, Moss DW. [ 3 2 ] M i t r a S K , Ve n k a t a r a g a n n a M V, S u n d a r a n R ,
Provisional recommendations on IFCC methods for the Gopumadhavan S. Protective effect of HD-03, a herbal
measurement of catalytic concentrations of enzymes. formulation against various hepatotoxic agents in rats. J
Clin Chem, 1977, 23(5): 887-899. Ethnopharmacol, 1998, 63(3):181-186.
[19] Bessey OA, Lowry O, Brock MJ. A method for the rapid [33] Rao SV, Padmavathi B, Reddy KS, Indira K. Protein
determination of alkaline phosphatase with five cubic metabolic response to carbon tetrachloride in a fresh
millimeters of serum. Biol chem, 1946:164, 321-29. fish, Sarotherrodon mossambicus. Chem Ecol, 1995, 11:
[20] Koller A. Proteins, In: Clinical Chemistry: Theory, 207-212.
analysis and co-relation, Kaplan LA, Pesce AJ, Eds. C. [34] Mohan H. The liver, biliary tract and exocrine pancreas.
V. Mosby, Toronto, 1984, 1268-1327. In: Text book of Pathology, 4th ed. New Delhi: Jaypee
[21] Doumas BT, Bayse DD, Carter RJ, Peters T, Schaffer R. Brothers Medical Publishers (P) Ltd, 2002, 22-224,

27
 Hepatoprotective activity of Capparis sepiaria / Asian Journal of Traditional Medicines, 2012, 7(1)

569-630. injury in primary cultured neonatal rat hepatocytes and

[35] Vijayan P, Prashanth HC, Vijayaraj P, Dhanaraj SA, in rats with hepatic damage. J Biomed Sci, 2006, 3(4):
Badami S, Suresh B. Hepatoprotective effect of the total 569-578.
alkaloid fraction of Solanum pseudocapsicum leaves. [37] Kim HJ, Chun YJ, Park JD, Kim SI, Roh JK, Jeong
Pharm Biol, 2003, 41(6): 443-448. TC. Protection of rat liver microsomes against carbon
[36] Yihang Wu, Fang W, Zheng Q, Lu L, Yao H, Zhou tetrachloride-induced lipid peroxidation by red ginseng
C. Hepatoprotective effect of total flavonoids from saponin through cytochrome P450 inhibition. Planta
Laggera alata against carbon tetrachloride-induced Med, 1997, 63(5): 415-418.

28
View publication stats