Available online at www.pharmresfoundation.com ISSN: 2229-3787 ...

Available online at www.pharmresfoundation.com ISSN: 2229-3787 

Journal of Advanced Pharmaceutical Research. 2011, 2(2), 64-75. 

Research paper 

Hepatoprotective activity of Clerodendron inerme (Verbenaceae) against acetaminophen induced 

hepatic injury in mice 

Ravindra kumar Chourasiya 1 , Siva Shankar Nayak 2,* 

1 Pharmaceutical Chemistry Research Laboratory, Department of Pharmaceutical Sciences, Dr. Hari Singh 

Gour University, Sagar, Madhya Pradesh, India. 

2 Department of Pharmaceutical Chemistry, College of Pharmaceutical Sciences, Mohuda, Berhampur, 

Orissa, India 

*Corresponding author E.Mail: sivanayak@yahoo.com 

Received: Feb 18, 2011; Accepted: May 22, 2011 

ABSTRACT 

The methanol and petroleum ether extracts of Clerodendron inerme (Verbenaceae) leaves were screened for its 

hepatoprotective activity in acetaminophen (250 mg/kg, i.p) induced liver damage in Swiss albino mice. The dose 

levels of 200, 300 mg/kg of both the extracts were used against the hepatotoxicity in liver of the animals. The 

methanol extract of C. inerme showed significantly (P

of phytoconstituents. Clerodendron inerme (L.) 

Gaertn (Family- Verbenaceae) is a scandent, 

straggling highly branched plant 0.9 - 2.1 m. long. 

The shrub is found in the wastelands, hedges, bank of 

rivers, sea and in various tropical parts of India 

(Kirtikar and Basu, 1998; Chatterjee and Pakrashi, 

1995; Chopra et al., 1956) and is acknowledged under 

vernaculars as ‘Glory Bower genus, Garden Quinine’ 

in English (Chourasiya et al., 2010); ‘Lanjai or 

Sangkupi’ in Hindi; ‘Cholora’ in Oriya; ‘Kundali, 

Samudrayuthika, Vanajai and Vanayuthika’ in 

Sanskrit. The poultice of the plant is used in buboes, 

vermifuge, anti periodic and as a substitute for quinine 

in remittent and intermittent fevers. Traditionally the 

leaves powder along with camphor, garlic or pepper 

was used for edema, muscular pains, rheumatic pains 

and roots were also used for venereal diseases 

(Nadkarni, 1996, Kirtikar and Basu, 1998; Chopra et 

al., 1956). Additionally, the fresh juices obtained from 

powdered leaves are used as alternative in scrofula; 

venereal diseases and is used orally in the treatment of 

jaundice (Chatterjee and Pakrashi, 1995). Alcoholic 

extract of leaves stimulates pregnant uterus, raises 

blood pressure and increases intestinal movements in 

rats. The main active chemical constituents include α- 

and β- amyrins, betulin, dehydroroyleanone, 

royleanone, neoclerodane (diterpenoids), Clerodermic 

acid, Cleroinermin, Acacetin, Apigenin (5-hydroxy-7, 

4’-dimethoxyflavone), Salvigenin (5-hydroxy-6, 7, 4’- 

trimethoxyflavone), Pectolinarigenin, Scutellarein and 

β-sitosterol (Chatterjee and Pakrashi, 1995; Rastogi 

and Mehrotra, 1995). The literature reveals that C. 

inerme is used orally in the treatment of jaundice in 

traditional Indian medicine and, moreover, the 

hepatoprotective activity of the stem and leaf water 

extract of C. inerme against carbon tetrachloride- 


induced liver toxicity was reported by Gopal and 

Sengottuvelu (2008). To date, no systematic study has 

been reported in an acute model regarding the 

Acetaminophen induced hepatotoxicity in mice. In the 

present study, the authors has been made to establish 

the hepatoprotective property of methanol and 

petroleum ether leaf extracts of C. inerme against 

Acetaminophen -induced hepatotoxicity in mice. 

MATERIALS AND METHODS 

Plant material 

The plant material (Leaves) used in the study. 

Clerodendron inerme (Verbenaceae) were collected in 

the month of August 2007 from the rural area of 

Mohuda in Ganjam District of Orissa, India. The plant 

was identified and authenticated by botanists Prof. S. 

K. Dash, HOD, PG Department of Bioscience, 

College of Pharmaceutical Sciences, Mohuda and 

comparing with the voucher specimen (CI-1) present 

in the herbarium, has been kept in the laboratory for 

experimental purpose. The collected plant leaves were 

washed and air-dried under the shade, cut into small 

pieces, powdered by a mechanical grinder and passed 

through 40-mesh sieve and stored in a closed vessel 

for future use. 

Preparation of C. inerme leave extracts 

The dried, powdered leaves of Clerodendron 

inerme (250 g) were extracted successively with 

petroleum ether 60–80°C (10 h) in soxhlet apparatus. 

A dark green colored petroleum ether extract was 

obtained. The same leave powder (marc), after proper 

air drying, again extracted with methanol (18 h) to 

produce a greenish brown semisolid mass. The 

extractions were carried out until the solvents become 

colorless. These extracts were further dried and 

concentrated by evaporating the solvent completely 

65

under vacuum at the range of boiling points of solvent 

(Methanol at 64°C and petroleum ether at 35°C) using 

rotary evaporator (Jain Scientific glass works, DTC 

201, Ambala cantt, India) to get crude extracts. The 

methanol extract (CIME) yield 15.7% w/w and 

petroleum ether extract (CIPE) yield 3.0% w/w 

respectively. The dried mass of the extracts were 

stored at 4°C and prepared an emulsion by triturating 

the accurately weighed quantity of the extract with 1% 

gum acacia and suspended in double distilled water at 

definite concentrations separately used for the 

pharmacological study. 

Chemical used 

Paracetamol (acetaminophen) was obtained 

from ALPA Laboratories, Indore, India and silymarin 

was obtained from Micro labs, Hosur, India as free 

sample, The assay kits used for the estimations of 

alanine aminotransferase (ALT), aspartate 

aminotransferase (AST), alkaline phosphatase (ALP), 

total bilirubin were purchased from (AGAPPE 

Diagnostics Pvt. Ltd., Maharashtra, India). All 

extractive solvents are used as analytical grade 

reagents (AR) and purchased from S.D. Fine 

Chemicals, Mumbai, India. 

Preliminary phytochemical analysis 

The methanol (CIME) and petroleum ether 

(CIPE) extracts were subjected to preliminary 

phytochemical screening for detection of major 

chemical groups. In each test, 10% w/v solution of the 

extracts were used and unless otherwise mentioned in 

individual test (Patil, 2001). 

Experimental animals 

All animal procedures were in strict 

accordance with the CPCSEA Guidelines (Committee 

for the purpose of control and supervision of 

experiment on animal). Healthy Swiss albino mice of 


7-8 weeks old weighing 20-35g of either sex were 

used. They were obtained from Jawaharlal Nehru 

Cancer Hospital and Research Centre, Bhopal (M.P) 

India. The mice were housed at a temperature of 20– 

25 0 C under a 12-h light/dark cycle with 50% of 

relative humidity. They were fed with standard animal 

feed e.g. Altromin pellets (Lipton’s India Ltd) and 

water ad libitum. This study complied with current 

ethical regulations on animal research in Jawaharlal 

Nehru Cancer Hospital and Research Centre, Bhopal 

and all mice used in the experiments were treated 

humanely. The use of animal was as per CPCSEA 

norms (CPCSEA Registration No. - 

500/01/a/CPCSEA/2001). Approval for experimental 

work was as per ethical committee of Jawaharlal 

Nehru Cancer Hospital and Research Centre, Bhopal 

M.P., India (Ref. No– 670/225-IAEC/2008/Project 

No.-51) .Animals was acclimatized to their 

environment for one week prior to experimentation. 

Acute Toxicity analysis 

Toxicity studies of the CIME and CIPE were 

performed to get the information, how safe is this 

extract for the therapeutic use. The LD50 value of 

CIME and CIPE were derived by the method of 

Litchfield and Wilcoxon 1949. The maximum non- 

lethal doses of both the extracts were found to be 3000 

mg/kg body weight, orally. The 1% gum acacia was 

used as a vehicle and showed no mortality. The 

determination of acute toxicity by adopting fixed dose 

the guideline of CPCSEA and 1/10 th of LD50 cut off 

values (Patil, 2001; Shivakumar, 2007) of the extracts 

were taken as screening dose. i.e. 200, 300 mg/kg for 

subsequent studies (CIME 200, 300 mg/kg and CIPE - 

200, 300 mg/kg). 

Acetaminophen -induced hepatotoxicity in mice 

(Acute model) 

66

Animals were randomized and divided into 

seven groups (1-7) of six mice in each group. Animals 

of Group-1 (control) received only vehicle (1 % Gum 

Acacia in distilled water). Animals of Group- 2 

(Paracetamol-control) received acetaminophen 250 

mg/kg only. Animals of Group-3 fed with 

acetaminophen 250 mg/kg and the standard drug 

Silymarin 50 mg/kg. Animals of Group-4 and 5 were 

fed with a single dose of acetaminophen 250 mg/kg 

and CIME of 200, 300 mg/kg. Similarly, animals of 

Group-6 and 7 were fed with a single dose of 

acetaminophen 250 mg/kg and CIPE of 200, 300 

mg/kg. Plant extracts were administered three hours 

after the administration of acetaminophen. All the 

treatments were done by means of a gavage or gastric 

tube. The treatments were continued for seven 

consecutive days and on the eighth day of the 

experiment, animals were anaesthetized with ether 

and blood sample was collected by puncturing the 

retro orbital plexus and serum was separated by 

centrifugation at 1500 rpm for 10min. The serum was 

then estimated for alanine aminotransferase (ALT), 

aspartate aminotransferase (AST), alkaline 

phosphatase (ALP) and total bilirubin level using 

assay kits according to the supplier (AGAPPE 

Diagnostics Pvt. Ltd., Maharashtra, India) 

specifications (Sabir and Rocha 2008). The estimation 

was done by using a photoanalyser instrument (FT-2; 

AMS); each determination was carried out with a 

suitable kit (AGAPPE Diagnostics Pvt. Ltd., 

Maharashtra, India) containing the necessary reagents 

at a predetermined wavelength for each parameter. 

Histopathological studies 

One animal belongs to the treated groups 

showing maximal activity as indicated by exhibit 

biochemical parameters from each test, positive 


control, hepatotoxin and control groups were selected 

for this purpose. The animals were sacrificed by ether 

anaesthesia and the abdomen was cut open to remove 

the liver, (5mm thick pieces) of the liver were treated 

in Bouin’s solution (mixture of 75 ml of saturated 

picric acid, 25 ml of 40% formaldehyde and 5ml of 

glacial acetic acid) for 12 hours, then embedded in 

paraffin wax and cut into 5 μm thick sections by using 

microtome and stained with haematoxylin-eosin dye 

and mounted in diphenyl xylene. Then the liver 

sections were observed under microscope for 

histopathological changes in liver architecture and 

photo microscopic were observed (100X) including 

necrosis, steatosis and fatty change of hepatic cells 

(Shai 2001; Saeed 2002). 

Statistical Analysis 

The results were recorded as Mean ± S.E.M. 

and statistical significance between treated groups 

with a control group was evaluated by One-way 

analysis of variance (ANOVA) followed by Dunnett’s 

t-test (Woodson, 1986). 

RESULTS 

Preliminary phytochemical analysis 

Results of different chemical tests on the 

CIME showed the presence of phytoconstituents, i.e., 

reducing sugars, flavonoids, phenolic compounds, 

alkaloids and gums; whereas CIPE showed a positive 

test for the presence of steroids and terpenoids. 

Effect of treatment with acetaminophen on serum 

ALT activities 

The estimation of all the biochemical 

parameters were carried out by using a photoanalyser 

(FT-2; AMS); serum ALT determination was done 

with a suitable kit (AGAPPE Diagnostics Reagent) 

containing the necessary reagents at a pre-determined 

67

wavelength 340 nm where as the rate of oxidation of 

NADH was measured kinetically by monitoring the 

decrease in absorbance at 340 nm (Murray, 1994). 

Mice were treated with paracetamol in group 2 alone 

developed significant liver cell damage as was evident 

for a significant (P < 0.001) increase in serum alanine 

aminotransferase enzyme activities. The oral 

administrations of CIME and CIPE at the different 

dose levels of 200 and 300 mg/kg (groups 4 to 7), 

were shown the lower significantly (P < 0.01) 

activities of marker enzymes and comparable to the 

group 3 of standard drug silymarin (50 mg/kg; P < 

0.01) as shown in Table 1. 

Effect of treatment with acetaminophen on the 

serum AST activities 

In the paracetamol treated group 2; there was 

a significant (P < 0.001) increase in aspartate 

aminotransferase activity compared to the normal 

group 1. AST activity of CIME and CIPE groups of 

two different doses 200 and 300 mg/kg (groups 4 to 7) 

significantly (P < 0.01) recovered the level of enzyme 

activities which is comparable with standard drug 

silymarin group-3 (P < 0.01) as shown in Table 1. 


levels of ALP activities 

The level of ALP activities were carried out 

using the kit AGAPPE Diagnostics Reagent analyzed 

in a photoanalyser (FT-2; AMS). By using a specific 

reagent at pH 10.3, alkaline phosphatase (ALP) 

catalyses the hydrolysis of colorless p-nitro phenyl 

phosphate (p-NPP) to yellow colored p-nitro phenol 

and phosphate. Change in absorbance due to yellow 

color formation is measured kinetically at 405 nm and 

is proportional to ALP activity in the sample (Moss 

and Henderson, 1994). Mice were treated with 

paracetamol (group 2) alone developed significant 


liver cell damage as was evident from a significant (P 

< 0.001) increase in serum enzyme levels of ALP 

compared to the normal group 1. CIME and CIPE 

groups of two different doses, 200 and 300 mg/kg 

significantly (P < 0.01 group 4, 5, 7; P < 0.05 group 

6), recovered the ALP level as compared to the 

standard drug silymarin group 3 (50 mg/kg; P < 0.01) 

as shown in Table 1. 


serum bilirubin levels 

Serum bilirubin levels in paracetamol-induced 

liver damage were estimated using an analytical kit 

from AGAPPE Diagnostics Reagent. The assay 

principle is based on the fact that total bilirubin reacts 

in the presence of caffeine with diazotized sulfanilic 

acid to form azobilirubin. The determination of direct 

bilirubin at 546 nm is proportional to the 

concentration of bilirubin (Willard and Meites, 1982). 

A significant (P < 0.001) increase in serum bilirubin 

level was seen in paracetamol-treated animals in 

group 2 compared to that of the normal group 1. 

Bilirubin levels for CIME and CIPE groups of two 

different doses 200 and 300 mg/kg significantly (P < 

0.01 group 4, 5, 7; P < 0.05 group 6) recovered 

against the paracetamol group 2 and both doses were 

comparable with the standard drug silymarin at 50 

mg/kg (P < 0.01) as shown in Table 1. 

Histopathological studies 

Histopathological examination of liver 

sections of control group 1 and standard group 3 

showed normal cellular architecture with distinct 

hepatic cells, sinusoidal spaces and central vein (Fig. 

1 and 3). Disarrangement of normal hepatic cells with 

centrilobular necrosis, vacuolization of cytoplasm and 

fatty degeneration were observed in the paracetamol 

treated mice of group 2 in Fig. 2. 

68


Table 1: Hepatoprotective Activity of C. inerme extracts on acetaminophen-induced Hepatotoxicity in mice. 

Serum biochemical parameter 

Groups Treatment 

AST 

ALT 

ALP Total bilirubin 

IU/L 

IU/L 

IU/L 

mg/dl 

Group 1 1 % Gum Acacia 49.50±1.36 65.33±1.40 83.42±1.75 2.23±0.08 

Group 2 Acetaminophen 

( 250 mg/kg) i.p. 

Group 3 Acetaminophen 250 mg/kg 

i.p. + Silymarin (50 mg/kg) 

a, b 

62.67±1.03 

a, b 

70.25±1.87 

a, b 

113.17±1.30 

a, b, c 

2.72±0.12 


i.p. CIME (200 mg/kg) 

a, b 

93.00±1.74 

a, b 

193.75±1.71 

a, b 

401.50±1.33 

a, b, c 

0.28±0.024 


i.p. + CIME (300 mg/kg) 

a, b 

78.25±1.40 

a, b 

151.50±1.60 

a, b 

367.25±2.04 

a, b, c 

0.25±0.019 


i.p. + CIPE (200 mg/kg) 

a, b 

110.75±1.75 

a, b 

196.25±1.66 

a, b 

486.75±1.02 

a, b, c 

0.29±0.017 


i.p. + CIPE (300 mg/kg) 

a, b 

76.75±1.17 

a, b 

179.50±1.90 

a, b 

397.50±2.02 

a, b, c 

0.26±0.019 

a b c 

P < 0.001, P < 0.01 and P < 0.05. 

All the values are expressed as Mean ± SEM.; n =6. The data was analyzed by evaluated by One-way analysis of variance 

(ANOVA) followed by Dunnett’s t-test, Group-2 compared with normal group-1 with a P < 0.001. Experimental groups from-3 to 

7 compared with group-2 with b P < 0.01 and c P < 0.05. 

158.50±1.33 a 

The liver sections of the mice treated with 

CIME (200 and 300 mg/kg) of groups 4 and 5 showed 

signs of protection as was evident by the absence of 

necrosis and vacuoles (Fig. 4 and 5) while the liver 

sections of the mice treated with CIPE (200 and 300 

mg/kg) of groups 6 and 7 showed reduction in fatty 

degeneration and absence of necrosis (Fig. 6 and 7). 

Fig. 1: Histopathology of liver of Negative control 

group. T.S. of liver revealed normochromatic 

hepatocytes (H), occasionally apoptosis, cart wheel 

nucleus (CWN) and lipid vacuoles (CV). A good mitotic 

index has been observed. H and E, 100x. 

129.42±1.76 a 

257.83±1.27 a 

4.02±0.20 a 

Fig. 2: Histopathology of liver of Positive control group. 

T.S. of liver revealed hyper chromatic and highly 

damaged hepatocytes (H). Large numbers of lipid 

vacuoles (LV) have been observed which propelled 

nucleus to the periphery of the cell. Frequent apoptosis 

(Apop) and necrosis (CWN) with poor mitotic index 

have been observed. H and E, 100x. 

DISCUSSION 

In acute toxicity study, oral administration of 

both CIME and CIPE did not produce any mortality in 

mice up to a dose level of 3 g/kg body weight. This 

may be due to broad non-toxic range of the plant, 

69

where the plant extract showed a high LD50 and 

relatively safety. Paracetamol (acetaminophen) is a 

widely used analgesic-antipyretic agent which is 

metabolized by the liver. 

Fig.3: Histopathology of liver of Standard group. T.S. 

of liver revealed normochromatic hepatocytes (H). 

Occasionally apoptosis, cart wheel nucleus (CWN) and 

lipid vacuoles (CV) have been observed with average 

mitotic index. H and E, 100x. 

Fig. 4: Histopathology of liver of Test group-1 

(Methanol Extract 200mg/kg bw orally). T.S. of 

liver revealed normochromatic hepatocytes(H) 

with good mitotic index. Occasionally lipid 

vacuoles (CV) have been observed without 

apoptosis (Apop), necrosis and cart wheel nucleus 

(CWN). H and E, 100x 

Over dosing of paracetamol to rats is reported 

to decrease their sensitivity to its hepatotoxic effects, 

which are associated with oxidative stress (Meredith, 

1996). So the studies on antioxidant enzymes, 


(Glutathione peroxidase (GPx), glutathione-S- 

transferase (GST), superoxide dismutase (SOD) and 

catalase (CAT) have been found to be of great 

importance in the assessment of liver damage. A small 

fraction of the drug is subject to oxidation reactions 

catalyzed by cytochrome P-450 enzymes in the liver, 

resulting in the generation of hepatotoxic N-acetyl-p- 

benzo-quinoneimine. 

Fig. 5: Histopathology of liver of Test group-2 

(Methanol Extract 300mg/kg bw orally). T.S. of liver 

revealed normochromatic hepatocytes with good mitotic 

index. Occasionally lipid vacuoles(CV), cart wheel 

nucleus(CWN) and apoptosis(Apop) have been 

observed without necrosis. H and E, 100x. 

Fig. 6: Histopathology of liver of Test group-3 (Pet. 

Ether Extract 200mg/kg bw orally). T.S. of liver 

revealed hyper chromatic hepatocytes(H) with average 

mitotic index. Frequently lipid vacuoles (CV) and cart 

wheel nucleus(CWN) have been observed. Few 

apoptotic hepatocyts(Apop) have been observed. H and 

E, 100x. 

70

Fig. 7: Histopathology of liver of Test group-4 (Pet. 

Ether Extract 300mg/kg bw orally). T.S. of liver 

revealed hyper chromatic hepatocytes (H) with average 

mitotic index. Frequently lipid vacuoles (LV) and cart 

wheel nucleus (CWN) have been observed. Few 

apoptosis (Apop) with necrotic hepatocytes have been 

observed. H and E, 100x 

Exposure to high doses of acetaminophen 

results in increased levels of N-acetyl-p-benzo- 

quinoneimine. Normally, the toxic oxidation 

metabolites generated in the liver are converted into 

non-toxic metabolites excreted in urine via 

conjugation with GSH, which contains sulphydryl 

groups. However, the intake of high doses of 

acetaminophen limits the capacity of GSH to detoxify 

N-acetyl-p-benzo-quinoneimine and results in the 

consumption of liver GSH stores (Mitchell et al., 

1973; Savides and Oehme, 1983). Oxidative stress is 

reported to constitute a major mechanism in the 

pathogenesis of acetaminophen induced liver damage 

(Ozdemirler et al., 1994). The binding of NAPQI with 

cellular proteins leads to necrosis in liver (Dahlin et 

al., 1984), which subsequently alters the liver function 

tests (LFTs). 


Reactive oxygen species (ROS) and nitrogen 

intermediates, produced by hepatic parenchyma and 

non-parenchyma cells are believed to be important 

factors contributing to Acetaminophen induced injury 

(Michael et al., 1999). In addition, the anti oxidative 

enzymes such as, super oxide dismutase (SOD) and 

catalase (CAT) also play an important role in the 

modulation of acetaminophen induced oxidative 

damage and is a commonly and widely used 

analgesic/ antipyretic agent. Hepatotoxic dose of 

acetaminophen step up the normal levels of hepatic 

enzymes (Morazzoni et al 1995). 

In living systems, liver is considered to be 

highly sensitive to toxic agents therefore it is easier to 

analyze hepatotoxicity/protection for short duration 

research. The study of different enzyme activities such 

as alanine amino transferase (ALT), asparate amino 

transferase (AST), alkaline phosphates (ALP) and 

total bilirubin have been found to be of great value in 

the assessment of clinical and experimental liver 

damage (Vermulen et al 1992). In the present 

investigation it was observed that the animals treated 

with acetaminophen resulted in significant hepatic 

damage shown by the elevated levels of serum 

markers. These elevated serum hepatic profile reflects 

hepatic structural integrity or the architect of liver 

tissue. Assessment of liver function can be made by 

estimating the activities of serum ALT, AST, ALP 

and total bilirubin, which are enzymes originally 

present in higher concentration in cytoplasm 

(Manokaran et al., 2008). 

When there is hepatopathy, these enzymes 

leak into the blood stream which serves as an 

indicator for the liver damage (Nkosi et al., 2005).The 

abnormally high level of serum ALT, AST, ALP and 

total bilirubin content observed in group 2 in our 

71

study are the consequence of paracetamol-induced 

liver dysfunction and denotes the damage to the 

hepatic cells (Table 1). Treatment with CIME and 

CIPE at dose levels of 200 and 300 mg/kg reduced the 

enhanced level of serum ALT, AST, ALP and total 

bilirubin level which seemed to offer protection and 

maintain the functional integrity of hepatic cells. The 

elevated levels of serum marker enzyme activities 

such as ALT, AST enzyme levels are a direct measure 

of hepatic injury and they show the status of liver. 

The elevated levels of serum enzymes are 

indicative of cellular leakage and loss of functional 

integrity of cell membrane in liver. Thus the lowering 

of the enzyme content in serum is a definite indication 

of the hepatoprotective action of a drug. CIME and 

CIPE at a dose of 200 mg/kg (P< 0.01) and 300 mg/kg 

decreased significantly (P < 0.01) the activity of both 

AST and ALT as compared to the standard drug 

silymarin. Serum ALP and bilirubin levels are also 

related to the status and function of hepatic cells. 

Increase in serum ALP level is due to increased 

synthesis, in the presence of increasing biliary 

pressure (Moss and Butterworth, 1974). 

In the present study, CIME and CIPE at a 

dose of 200 mg/ kg (P < 0.01 and P < 0.05) and CIME 

and CIPE at a dose 300 mg/kg, were found to reduce 

significantly (P < 0.01) both serum ALP and bilirubin 

levels in the treated groups which are comparable to 

the standard drug silymarin (Table 1). 

Hepatoprotective activity of C. inerme was shown by 

its ability to inhibit paracetamol-induced liver damage 

in mice. The fall in serum marker level by CIME (300 

mg/kg) was significantly (P < 0.01) able to alter the 

toxic condition of the hepatocytes as compared to 

CIPE. 


The histopathological study has been done in 

liver tissue. Negative control group revealed normal 

hepatocyte with normochromatic cellular components 

(fig.1). The liver section of mice which was treated 

with paracetamol (Acetaminophen) revealed hypo- 

chromatic cells, apoptotic bodies with more 

intracellular lipid vacuoles which was more prominent 

for shifting of nucleus to the periphery. Cart wheel 

nucleus was also seen (fig.2). When the animal was 

treated with Silymarin there was a slight reduction of 

apoptotic cells and cart wheel nucleus (fig.3). On the 

other hand when the animal was administered with 

200 mg/kg bw orally CIME revealed good mitotic 

index, very few lipid vacuoles, cart wheel nucleus and 

very few apoptotic bodies. Section was suggestive of 

normochromatic cells (fig.4) when the animal was 

treated with 300 mg/kg bw orally CIME. It has been 

also observed that all the cells were normochromatic 

hepatocytes with good mitotic index, negligible lipid 

vacuoles were present and there was no evidence of 

cellular toxicity (fig.5). On the other hand, CIPE @ 

200 mg/kg b.w orally revealed hyper chromatic, 

dysplastic cells, distorted hepatocyte and large no. of 

lipid vacuoles with low mitotic index (fig.6). When 

animal was treated with CIPE @ 300 mg/kg b.w 

orally revealed hyper chromatic hepatocytes with 

average mitotic index (fig.7). A comparative 

histopathological study of livers from different 

experimental groups further corroborated the 

hepatoprotective efficacy of CIME and CIPE. 

Hepatoprotective activity of C. inerme was 

shown by its ability to inhibit paracetamol-induced 

liver damage in mice. The fall in serum marker level 

by CIME (300 mg/kg) was significantly (P < 0.01) 

able to alter the toxic condition of the hepatocytes as 

compared to CIPE at a dose level of 200 and 300 

72

mg/kg, so as to protect the membrane integrity against 

acetaminophen-induced leakage of marker enzymes. 

This means there was a potential functional change in 

the hepatocytes of the hepatic cells. Hence, it has been 

concluded that the methanol extract of C. inerme 

(CIME) at dose levels of 300 mg/kg revealed its 

potential significance as a hepatoprotection against 

acetaminophen-induced toxicity which was further 

supported by the presence of phytoconstituents such 

as flavonoids, polyphenols, and tannins. All the results 

can be comparable with the standard drug silymarin. 

Hence, we conclude that CIME and CIPE at higher 

dose revealed its potential significant as a 

hepatoprotection against Acetaminophen induced 

toxicity. However, a mild hepatic cell aberration was 

found in lower doses. A comparative 

histopathological study of liver from different 

experimental groups corroborated the 

hepatoprotective efficacy of methanol extract which 

was more effective than the petroleum ether extract of 

Clerodendron inerme. Further detailed studies may, 

however, confirm the utility profile of this drug. 

ACKNOWLEDGEMENTS 

Authors are thankfully acknowledged to 

Jawaharlal Nehru Cancer Hospital and Research 

Centre, Bhopal (M.P) India and College of 

Pharmaceutical sciences, Mohuda, Berhampur, India 

for providing research facilities. The authors also wish 

to thank Dr. N.Ganesh, Head, Department of 

Research, Jawaharlal Nehru Cancer Hospital and 

Research Centre, Bhopal (M.P) India for his valuable 

help and suggestions during the course of work and 

also thankful to the Prof. S. K. Dash, HOD, PG 

Department of Biosciences, College of 

Pharmaceutical sciences, Mohuda, Berhampur, for the 

identification of the plant. 


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