Journal of Cell and Tissue Research Vol.

9(3) 1981-1984 (2009)

ISSN: 0974- 0910 (Available online at www.tcrjournals.com) Original Article

EVALUATION OF THE PROTECTIVE EFFICACY OF WOODFORDIA

FRUTICOSA ON PHENYTOIN INDUCED LIVER
DAMAGE IN RATS
?
BRINDHA, D. AND GEETHA, R.

Department of Biochemistry, PSG College of Arts and Science, Coimbatore- 641 046, Tamil Nadu.
E. mail: brindhavenkatesh@ymail.com

Received: August 15, 2009; Accepted: September 15, 2009

Abstract: The present study was carried out to investigate the efficacy of hydroalcoholic extract
of flowers of Woodfordia fruticosa against phenytoin induced hepatic injury in albino rats. The
extract in the dose of 250mg/kg body weight was administered orally once daily for 21 days. The
hepatoprotective activity of the extract was assessed by measuring the total bilirubin, total
protein, total cholesterol, triglycerides and serum marker enzymes in control and experimental
animals. The results revealed significant reduction (p<0.05) in total bilirubin and serum marker
enzymes and increase in total proteins in the animals treated with the flower extract. However,
significant rise in these serum enzymes and decrease in total protein level was noticed in phenytoin
treated group indicating the hepatic damage, while the extract of Woodfordia fruticosa
significantly protected the phenytoin induced hepatocellular injury.

Key words: Hepatoprotective, Woodfordia fruticosa, Phenytoin

INTRODUCTION as foods, beverages and herbal plants, which can

Liver, the largest gland in the body is being evolved
to maintain the body’s internal milieu and also protect
itself from the challenge it faces during its function
[1]. It is concerned with almost all the biochemical
pathways related to growth, fight against disease,
nutrient supply, energy provision and reproduction [2].
Liver, particularly, is vulnerable to drug induced
toxicity mainly because of its role as a primary organ
of drug elimination and its subsequent exposure to
potential toxins [3,4]. Hepatotoxins can affect
biological macromolecules such as proteins, lipids,
RNA, DNA and induce several types of lesions, of
which genotoxic alterations may lead to
carcinogenesis [5]. In order to intervene in the earlier
stages of chemical mediated toxicity, use of chemo-
preventive agents may be considered as an effective
alternative. Among various agents with chemop-
reventive effects, phytoproducts are gaining more and
more attention gradually due to their decreased
toxicity and high efficacy [6], including those used
either increase the efficacy of the antioxidants present
in our body [7] or possess the intrinsic antioxidant
activity. Herbal plants have been found beneficial to
humans as they are the source of various medicinal
compounds and some of them possess antioxidant
property [8-12].
Woodfordia fruticosa belongs to the family
Lythraceae, popularly known as Dhataki and is a
well known non-wood forest produce that is a much
used medicinal plant in Ayurvedic and Unani systems
of medicine in India [13]. The plant Woodfordia
fruticosa is a potential candidate for peptic ulcer
disease [14]. According to the Indian system of
medicine, the flower is pungent, acrid, cooling, toxic,
alexiteric, uterine sedative and anthelmintic and is
useful in thirst, dysentery, leprosy, erysipelas, blood
disease, leucorrhoea, menorrhagia and toothache
[15]. Oil based flower extract has always been
recommended for open wounds [16]. The major
compounds identified in Woodfordia fruticosa are

1981
 J. Cell Tissue Research

predominately phenolics, particularly hydrolysable Group III received hydroalcoholic Woodfordia

tannins and flavonoids; another constituent isolated
was woodfordin C with antitumor propety [17,18].
Literature survey revealed that, this plant has not
been subjected to pharmacological screening for its
hepatoprotective activity. In view of this fact, the
present investigation has been made to evaluate the
hepatoprotective activity of Woodfordia fruticosa
against phenytoin induced hepatic dysfunction in rats.
MATERIALS AND METHODS
Plant material: The dried flowers of Woodfordia
fruticosa were procured from the local market in
Coimbatore. The sample was identified and
authenticated by the botanist, Department of Botany,
PSG College of Arts and Science, Coimbatore.
Extraction: About 70g of dried plant material was
extracted with 300 ml of 50 % alcohol by using a
separating funnel with occasional shaking for 72 hrs.
The hydroalcoholic extract of the plant material was
prepared by Soxhlet extraction. Then the extract was
tested for hepatoprotective activity at a concentration
of 250 mg/kg body weight.
fruticosa extract 250mg/kg body weight of animals.
Group IV received Phenytoin suspension 250 mg/kg
bw + hydroalcoholic Woodfordia fruticosa extract
250mg / kg body weight of animals. Group V
received Phenytoin suspension 250 mg/kg + silymarin
100 mg/kg body weight for 21 days. All the treatments
were given orally by means of a gastric tube.
At the end of the treatment, blood samples of all
animals were collected in sterile centrifuge tubes and
allowed to clot. Serum was separated and used for
the assay of protein [20], bilirubin [21], lipid profiles
cholesterol [22] and triglycerides [23], serum marker
enzymes viz., aspartate amino tranferase , alanine
amino transferase [24], alkaline phospatase [25],
lactate dehydrogenase [26] and gamma glutamyl
tranferase [27].
Statistical Analysis: All the data were expressed
as mean ± SD from six animals of each group. The
statistical significance was evaluated by one-way
analysis of variance (ANOVA) using SPSS version
9.0 (SPSS, Cary, NC, USA). Values were considered
statistically significant when P<0.05.

Experimental animals: Male Wistar albino rats, 6 RESULTS

to 8 weeks of age and weighing 150-200 g, were
used. The animals were kept in clean and dry plastic
cages, with 12h:12h light-dark cycle at 25 ± 2 ºC
temperature and 45 - 55 % relative humidity. The
animals were fed with a commercial diet (Amrut
Feed: Mumbai) and water ad libitum. The experim-
ental protocols were approved by the Institutional
animal ethics committee [19].
Experiment design: Male Wistar albino rats (30
animals) were divided into five groups comprising
six animals in each group. Group I (control) received
only distilled water. Group II (positive control)
received 250 mg/kg bw Phenytoin suspension only.
Data pertaining to the levels of biochemical paramet-
ers are presented in table 1 and 2. The hepatic
enzymes viz., aspartate amino transferase, alanine
amino transferase, lactate dehydrogenase and gamma
glutamyl transferase in serum significantly (p<0.05)
increased in phenytoin treated animals as compared
to control. The plant extract treatment significantly
reversed the levels of all enzymes in group III and
group V animals. All the parameters were under
normal limits in the silymarin treated group, which
acted as a positive control.
The total protein concentration of serum was lesser

Table 1: Effect of Woodfordia fruticosa flower extract against Phenytoin induced changes in the activities of serum hepatic
marker enzymes on control and experimental rats. Values are mean ± SD for 6 rats in each group. Values with different superscript
letter differ significantly at P < 0.05
Groups AST (U/L) ALT (U/L) ALP (U/L) GGT (U/L) LDH (U/L)
Control 55.83 ± 3.43 53.17 ± 3.06 124.50 ± 5.43 3.97 ± 0.41 147.83 ± 5.98
Phenytoin 329 ± 1.26a 312.67 ± 11.78a 237.67 ± 16.93a 8.87 ±0.50a 409.50 ± 13.46a
Woodfordia fruticosa flower extract 82.83 ± .12b 85.50 ± 3.94b 154.3 ± 8.31b 5.50 ± 0.62b 169.1 ± 5.31b
Phenytoin + Woodfordia fruticosa 75.83 ± 7.22 75.17 ± 4.45 143 ± 10.70 4.87 ± 0.42 159.67 ± 3.44
flower extract
Sylimarin 53.50 ± 5.68 51 ± 4.20 131.17 ± 8.52 4.15 ± 0.84 158.33 ± 2.94

1982
 Brindha and Geetha
Table 2: Effect of Woodfordia fruticosa flower extract against Phenytoin induced changes on Bilirubin, Cholesterol, Triglycerides
and Protein in control and experimental rats. Values are mean ± SD for 6 rats in each group. Values with different superscript letter
differ significantly at P < 0.05
Groups Bilirubin (mg/dl)
Control 0.42 ± 0.04
Phenytoin 1.59 ± 0.11 a
Woodfordia fruticosa flower extract 0.66 ± 0.05 b
Phenytoin + Woodfordia fruticosa 0.55 ± 0.04
flower extract
Sylimarin 0.68 ± 0.05
in phenytoin treated group when compared with
normal control and it attained an almost normal value
in the hydroalcoholic flower extract treated animals.
The level of total bilirubin and lipid profiles namely
cholesterol and triglycerides in serum recorded
significant increment in rats treated with phenytoin
when compared with normal controls. The
administration of the flower extract and silymarin
significantly reduced the level of all above mentioned
biochemical indicators.
DISCUSSION
The liver is a vital organ and can be injured by number
of chemicals and drugs. In the present study,
phenytoin was selected as hepatotoxicant to induce
liver damage. Phenytoin, a hydantion anticonvulsant
is used widely in the treatment of seizure [28,29].
Phenytoin is metabolized by hepatic cytochorome
P450 enzymes to reactive aromatic intermediates
(arene oxides). These arene metabolites of phenytoin
may be involved in the pathogenesis of drug induced
hepatotoxicity [30,31]. The activities of serum marker
enzymes were significantly higher in phenytoin
treated rats as compared to the controls. These
results show that phenytoin is capable of affecting
both mitochon-drial and cytosolic enzymes. The
concurrent treatment of phenytoin with the flower
extract mediated restoration in the levels of serum
marker enzymes towards respective normal values
is an indication of stabilization of plasma membranes
as well as repair of hepatic tissue due to damage
caused by this toxicant.
The site specific oxidative damage of some of the
susceptible amino acids of protein is regarded as the
major cause of metabolite dysfunction during
pathogenesis. The capacity of liver to synthesize
albumin is adversely affected by hepatotoxins [32].
The lower level of total protein recorded in the serum
of phenytoin treated rats can be attributed to these
1983
Cholesterol (mg/dl) Triglycerides (mg/dl)
108.33 ± 14.72 169 ± 8.85
278.33 ± 56.01a 206.5 ± 22.11 a
170 ± 20 b 130 ± 19.44 b
153.33 ± 30.11 162.83 ± 20.88
128.33 ± 29.94 158.33 ±17.68
features. Attainment of near normalcy in protein
content of serum in flower extract treated rats further
confirmed the anti-hepatotoxic effect of W. fruticosa.
Bilirubin concentration has been used to evaluate
chemically induced hepatic injury. The various forms
of liver disorders showed hyperbilirubinemia [33].
Phenytoin intoxication in rats increased the level of
bilirubin content in serum which might be due to the
destruction of erythrocytes by toxic metabolite
leading to a failure, to extract bilirubin. Simultaneous
administrations of the flower extract with phenytoin
decreased the elevated bilirubin level.
The increased serum triglyceride levels in phenytoin
treated rats may be due to the decreased activity of
lipoprotein lipase, which is involved in the uptake of
triglyceride- rich lipoprotein by the extra hepatic
tissues [34]. Pretreatment with the flower extract
reduced the elevated cholesterol and triglyceride
levels, suggesting that the extract prevented pheny-
toin induced hyperlipidemia probably due to their
hepatoprotective activity. In all the parameters
studied, the hepatoprotective activity of W. fruticosa
was significant being similar to that of silymarin.
Herbal medicines are known for centuries to be
protective against one or the other diseases, the
pathophysiological bases of which were not known
in those periods and even to day majority of them
are unidentified. Woodfordia fruticosa is used in
the Indian systems of medicine like Ayurveda, Siddha
and Unani [35]. Plant constituents like tannins and
flavonoids are well known for their antioxidant and
hepatoprotective activities [36,37]. The flowers of
Woodfordia fruticosa contain substantially high
amount of tannins, flavonoids and some polyphenolic
compounds, that show antioxidant and hepatoprote-
ctive properties [38].
From overall data it is concluded that the flower
 J. Cell Tissue Research
extract of Woodfordia fruticosa exhibits antioxidant
and hepatoprtective properties and may be screened
further as liver protector. However, further studies
are needed to isolate and purify the active principal
involved in the hepatoprotective function.
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