Content uploaded by Thuwaini Mm
Author content
All content in this area was uploaded by Thuwaini Mm on Oct 09, 2018
Content may be subject to copyright.
Thuwaini et al. European Journal of Pharmaceutical and Medical Research
www.ejpmr.com
36
"HEPATOPROTECTIVE EFFECTS OF THE AQUEOUS EXTRACT OF CLOVE
(SYZYGIUMAROMATICUM) AGAINST PARACETAMOL-INDUCED
HEPATOTOXICITY AND OXIDATIVE STRESS IN RATS"
1Mahdi M. Thuwaini, 2*Mohammed Abdul-Mounther and 3Hanaa S. Kadhem
1Ph.D. Pathology, College of Nursing, Thi-Qar University.
2Ph.D. Biochemist. College of medicine, Missan Univ. Iraq.
3Ph.D., Physiology, College of Science, Biology Department / University of Basra.
Article Received on 26/05/2016 Article Revised on 15/06/2016 Article Accepted on 06/07/2016
INTRODUCTION
The liver is a key organ in the body and is the First
official for the metabolism of internal and external
agents. It plays vital role in drug disappearing and
detoxification and liver damage may be caused by
xenobiotics, alcohol consumption, malnutrition,
infection, anemia and medications (Mroueh et al.,
2004). Furthermore, the liver has been expected to be
especially susceptible to the noxious effects of
miscellaneous use and until now PCM hepatotoxicity
influences are still expostulative.
Various types of drugs such as acetaminophen,
chloroquine and isoniazid are inducers of hepatotoxicity
in world. More of the hepatotoxic chemicals damage
liver cells basically by inducing lipid peroxidation and
other oxidative damages (Hiraganahalli, et al., 2012).
Paracetamol (PCM) is acetaminophen or N-acetyl-p-
aminophenol (APAP), but a high dose can lead to
undesirable side effects, such as hepatotoxicity with
oxidative stress as one of the possible mechanisms
mediating the event. It was first discovered in 1889 and
is a widely used no prescriptive analgesic and antipyretic
agent (Brown, 1968). Paracetamol (PCM) is widely
used as analgesic and antipyretic drug, but at high
dose it leads to undesirable side effect s, such as
hepatotoxicity.Paracetamol metabolism depends on the
age and sex, after reception a therapeutic dose,
paracetamol tends to sulfation and glucuronidation and
outputs non-toxic metabolites with urine.
Paracetamol hepatotoxicity is caused by the reaction
metabolite N-acetyl-p-benzoquinoneimine (NAPQI),
which causes oxidative stress and glutathione (GSH)
depletion (Boyd and Bereczky, 1966). Where About 4%
of a therapeutic dose is metabolised by the cytochromes
P450, chiefly CYP2E1, to a potentially toxic
intermediate metabolite N-acetyl-p-benzoquinone imine
SJIF Impact Factor 3.628
Research Article
ISSN 2394-3211
EJPMR
EUROPEAN JOURNAL OF PHARMACEUTICAL
AND MEDICAL RESEARCH
www.ejpmr.com
ejpmr, 2016,3(8), 36-42
*Corresponding Author: Mohammed Abdul-Mounther
Ph.D. Biochemist. College of medicine, Missan Univ. Iraq.
ABSTRACT
Hepatoprotective activity of aqueous extract of clove (Syzygiumaromaticum) in albino rats was observed against
controversial hepatotoxicity effects of paracetamol(PCM) induced liver toxicity in rats. Paracetamol was
administered to induce hepatic damage in Wistar albino rats. 100 and 200 mg/kg doses of clove extract and were
used as treatment groups. In present study, the effects of clove extract [100 mg per kg B.W. and 200 mg/kg.B.W.]
as alone. On PCM-induced hepatotoxicity were examined. Rats were divided haphazardly into six groups
containing 6 rats each. The control group received distal water (D.W.). Other groups were treated with PCM
(600mg/kg) alone, (600mg/kg PCM + 100mg/kg clove extract), and (600mg/kg PCM + 200mg/kg extract)
respectively for 4 weeks. The blood samples were analyzed for biochemical labels of hepatic injury and tissue
samples were subjected for assessing of liver antioxidants and histopathological studies. Analysis of the treated rats
with PCM (600 mg/kg) showed Paracetamol induced male rat hepatotoxicity represented by significant decline in
the serum total albumin (P< 0.05). Conversely, the study declared significantly increment (P< 0.001), bilirubin,
ALT, AST and ALP as shown in group2 (induction group) in compared with group1 (control group). While,
simultaneous administration of clove aqueous extract (100mg/kg and 200mg/kg) with paracetamol, was displayed
significantly attenuated the adverse changes in the serum total albumin, bilirubine, ALT IU/L, AST, and ALP. The
histopathological examination in the liver of rats also encouraging that clove extract markedly diminished the
toxicity of PCM and keeps the histoarchitecture of the liver tissue to near normal. Hence, the results postulate that
clove extract acts as a potent hepatoprotective agent against PCM induced hepatotoxicity in rats.
KEYWORDS: Hepatotoxicity, Syzygiumaromaticum, Wister rats, Paracetamol, Serum liver enzymes.
Thuwaini et al. European Journal of Pharmaceutical and Medical Research
www.ejpmr.com
37
(NAPQI). The highest concentration of CYP2El is
located in centrilobular hepatocytes around the central
vein and reflects the primary hepatic injury produced by
paracetamol. Under normal conditions and therapeutic
doses, NAPQI integrates with intracellular glutathione to
become a non-toxic mercapturate derivative with urinary
excretion. However, after taking a high dose, the
normally minor CYP2E1 pathway becomes important.
When the production of NAPQI surpass the capacity to
detoxify it, the increase NAPQI binds to cellular
components, causing mitochondrial damage and
eventually the death of the hepatocyte. If a
sufficientadequate dose is taken, hepatocyte death may
be massive and produce acute liver failure (Basu, et al.,
2014; Heard, et al., 2012).
Today, herbal medicine is becoming more and more
popular as a safety and effective means of treatment for
many different medical conditions. Herbs are often
favored because they are natural and do not put noxious
chemicals into the body. Herbs are available fresh, as
capsules, powders, extracts, roots, etc (Dhinahar, et al.,
2011).
However, usually that the use of aromatic plants hardly
be higher in popular medicine, which had previously
been documented their characteristics which include:
anti-bacteria, fungi and yeasts (Karkosh, 2012). Most of
those properties of these plants have able to control
microorganisms related to skin diseases,. In addition to
tooth decay and food rotting (Chaieb et al., 2007).
In any case, during the past few years there was observed
an exponential growth in the field of herbal medicament
that has acquired wide popularity medicines due to
natural sources and the lack of side effects. Currently it
has been evaluated natural diverse products to its
protection of hepatic different patterns of hepatotoxicity
(Grover , et al., 2002; Madkour, et al., 2013).
Moreover, clove is one of the most valuable spices used
for centuries as a preservative for food and many medical
purposes. The origins of this type of original spices in
Indonesia, either at the present time are grown in
different regions of the world (Cortés-Rojas et al., 2014).
Where half of the world production of cloves used in
Indonesia to kretek cigarettes by one part of the clove
mixture with two parts of tobacco. Where the their
benefit were obtained through the flower and bud as an
anti-viral and antibacterial, libido exciting, high blood
pressure, carminative, it's main prefer component is
eugenol (de Paoli et al., 2007; Politeo et al., 2010; Koba
et al., 2011).
Furthermore, Clove buds products more than 15 to 20%
of a volatile oil that is accountable for the Distinctive
smell and flavor. The main components of clove bud oil
distilled (60-90%) are phenylpropanoids, including in the
first place eugenol (4-allyl-2-methoxyphenol) and
carvacrol, thymol and cinnamaldehyde and carvacrol,
thymol and cinnamaldehyde (Chomchalow, 1996).
Clove is described as a repellent for fever, disinfectant
and sterilizer for the stomach, heals sores and pain of the
head and protects from epidemics and helps digestion
and calms dental pain and reduces inflammation and
allergy alerts the heart and stomach. And it is extracted
from the seeds of clove oil pilot called eugenol, which
has a special dental pain therapeutic properties, and can
also extract the oil from the clove tree and its leaves
market by distillation (Nosrati, et al., 2011; Djilani,
2012).
Furthermore, Clove is one of the wealthiest resources of
phenolic compounds and it has big potential for
pharmaceutical, cosmetic, food and agricultural
applications [Diego, et al. 2014]. Since flavonoidsd are
capable to decline xenobiotic-induced hepatotoxicity in
animals and repel the damaging influences of oxidative
stress, collaborating with natural systems like
endogenous protective antioxidant enzymes, clove
appears antioxidant peculiarities and its extracts could be
used as food antioxidants [Kadarian, et al., 2002; Diego,
et al., 2014].
However, commonly use of liver function tests in order
to monitor for liver disease, to observe the development
of hit disease and monitor the effects of these properties
of potentially hepatotoxicity drugs (Muriel, et al., 1992).
Increment the levels of AST and ALT are an indicator of
cellular leakage and functional disorder of the liver cell
membrane(AbolfathiMohajeri, et al., 2012). ALP is
membrane cohesion and its changing is likely to be
affecting the permeability of the membrane and
disturbances in the transfer of outputs (Mehana, et al.,
2012). On the other hand, albumin and bilirubin values
are connected with the excretive and synthetic functions
of hepatic cells (Muriel, et al., 1992).
The goal of this deliberating was carried out to examine
the anti activity of clove extraction against
heptatotoxicity with regard to its effects on liver cells;
histopathology, routine liver function tests.
MATERIAL AND METHODS
Animals
Male Wister albino rats (Rattusnorvegicous) were
obtained from animal center of Thi-Qar university
\college of science/ Iraq, weighing 250 ± 10 g, age (6-80)
months, The animals were kept in standard conditions
(23 ± 2 °C, 12 h light / dark cycle). Standard diet and
water were given ad libitum.
Drug and plant extract
Paracetamol drug (SDI Co, Iraq) were dissolved in
normal saline before use. While Clove dried flower buds
were purchased from the local market, Iraq.
Thuwaini et al. European Journal of Pharmaceutical and Medical Research
www.ejpmr.com
38
Kits for Biochemical Analysis
Kits for Biochemical Analysis Diagnostic commercial
kits for biochemical analyses were purchased from (cat
no. 80014, France, and (cat no .al 146 and cat no. AS
147, united kingdom).
Experimental Design and assembling of rats
Paracetamol (PCM) hepatotoxicity was induced in three
groups of rats. Where, the animals were divided
randomly into six groups (6 each), the first group was
given normal saline (the vehicle) 0.5ml / animal /daily
/I.P.,for 4wks, to serve as (negative) control group. The
second group (induction) or positive group was given
paracetamol (PCM) 600mg/kg/ orally (P.O), alone, three
times weekly for 8 weeks. The third and fourth groups
were received clove extract (100mg/kg/ day and
200mg/kg/ day) alone respectively for 8 weeks as a
single daily (P.O.), while the animals in fifth and six
groups were received (600mg/kg PCM + 100mg/kg
B.W. clove extract), and (600mg/kg PCM + 200mg/kg
clove extract) respectively, to the end of the experiment.
At the end of the treatment period, all animal were killed
by neck dislocation after light anesthesia with diethyl
ether. Blood samples were taken by cardiac puncture.
The collected blood samples were permitted to clot. Sera
were removed by centrifugation at 3000 rpm for 5 min.
and stored at -20°C until analyzed.
Biochemical assay
Liver function tests were carried out by an enzymatic
assay (Drotman and Lawhorn, 1978). Where Serum
blood samples were analyzed for the activities of
Aspartate Transaminase (AST), Alanine transaminase
(ALT), alkaline phosphatase (ALP) and for concentration
of albumin and bilirubin. using a spectrophotometric
autoanalyzer (Olympus AU-2700).
Histopathological examination
Necropsy was conducted to identify gross lesion, after
anesthetizing, the rats were dissected. The livers were
removed and cleaned. Liver tissue samples were fixed in
10% formalin for24 and then washed –dehydrated-
cleared and the organ was embedded in paraffin(Izbicki,
2002), sections about 4μm diameter was cutting and
stained with hematoxylin-Eosin (H and E) for
microscopic examination(Andrew, et al., 2008).. The
histological changes were assessed by observation of
lesions microscopically.
Statistical analysis
Statistical Package (SPSS, version 16) was done using
student T-test for the analysis of the data. Furthermore,
to determine the significance among groups. Where Data
were expressed between means (Mean±Standard
deviations (Mean±SD). Namely,the results in all above
were accepted as statistical significant when the (p) value
less than (p< 0.05).
RESULTS
As shown in table1, the results of hepatoprotective
activity of clove aqueous extract and The hepatotoxicity
of paracetamol(PCM) in rats, demonstrated that the
effects whether via clove aqueous extract or PCM on
serum levels of albumin, bilirubin and liver enzymes
(ALT, AST and ALP) in rats. Where paracetamol
hepatotoxicity was correlated with significant decrease in
the serum albumin (mg/dl) P< 0.05. Conversely, this
study was appeared significantly increased (P< 0.001) in
bilirubin (mg/dl), ALT, AST and ALP as appear in
group2 (induction group) in comparison with group1
(control group). But when using of extract alone in
healthy rats (group3 and 4) caused little insignificant
changes in serum total bilirubin, albumin, ALT, AST and
ALP. Anyhow, immediate administrations of extract
(100mg/kg and 200mg/kg) with paracetamol (600mglkg)
displayed significant decreased the adverse changes in
the serum total bilirubin, ALT, AST and ALP (P< 0.01).
While, Concomitant oral administration of extract
(100mg/kg and 200mg/kg) with paracetamol (group 5
and 6) showed significant increment (P<0.05) in the
mean values of serum total albumin. In Compared with
paracetamol -treated rats. However, it didn't repeat them
to the normal limits.
Histopathologicalexamination of the liver sections in
current study gets from each group. photomicrograph of
the liver section obtained from groups treated either with
normal saline (vehicle) or PCM administration and rats
that were treated at same time with clove extract (two
doses). The control group declares normal cellular
architecture with identified hepatocytes structure,
sinusoidal spaces and central veins, with normal radial
arrangements around central vein (CV) (Figure 1a). In
the meantime, grievous histopathological changes were
clearly noticed in PCM-intoxicated rats revealing
centrilobular hepatic necrosis, with occasional plotted
areas of moderate necrosis. Pronounced grade of fatty
changes and marked centrilobular hepatic infiltration of
lymphocytes and nuetrophils were obviously observed,
congestion in portal vein (PV) with fibrous tissue
proliferation, extended from portal area and forming
portal bridging fibrous septae (Fig. 1b), besides, showing
ballooning degeneration of hepatocytes associated with
vacuolar changes (Fig.1c).When clove extract was
treated simultaneously withPCM administration,
significant decreases of hepatocellular changes were
observed. In addition, showing closer to normal liver
structure (Figure 1 c). Identical observation was seen
when clove extract alone was administered to the rats.
Thuwaini et al. European Journal of Pharmaceutical and Medical Research
www.ejpmr.com
39
Table1: Effect of administration of low dose (10mg/kg/day) and high dose (20 mg/kg/day) of clove extract on
Paracetamol induced hepatotoxicity in rats. Values are expressed as Mean ± SD.
Groups
Bilirubin mg/dl
Albumin g/dl
ALT U/L
AST U/L
ALP U/L
G1
0.61±0.01
5.02±0.77
55.29±4.32
69.81±8.44
148.91±13.70
G2
1.51±0.11 *
3.1±0.66 *
181.90±14.50*
198.60±17.80*
238.23±18.02 *
G3
0.59±0.01NS
4.88±0.05 NS
54.47±4.62 NS
72.61±7.43 NS
145.95±12.02NS
G4
0.66±0.05 #
5.04±0.10 #
56.85±7.32#
68.62±13.12#
144.92±14.25#
G5
1.01±0.04 #
4.51±0.09 #
126.36±7.32 #
141.63±11.60#
201.31±13.14 #
G6
0.99±0.11 #
4.59± 08 #
121±9.01 #
128±6.61 #
198.05± 13.33#
* Significant change (P< 0.05).compared to control vehicle group.
#: Significant change (P< 0.05) compared to induction group.NS: Non-significant
A. Healthy rats B. Inducted Rats
C. Inducted Rats D. PCM rats treated with Extract
Figure 1: a- Histopathology of healthy rat liver declaringnormal histology of hepatic structure. B–c. Inducted
rats liver showing intensity fatty changes, sinusoidal feathery degeneration and necrosis, congestion in portal
vein (PV) with fibrous tissue proliferation, ballooning and sever degeneration of hepatocytes d. healthy rat liver
treated with clove extract plus PCM, showing mild fatty change, mild sinusoidal dilation and congestion viz
closer to normal hepaticstructure. (H&E 200X).
DISCUSSION
Liver regulates many important functions including
metabolism, and is an important organ for detoxification
of the liver, which can cause damage to the liver during
the metabolic interaction. Pathogenesis of this damage
includes all types of cells in the liver by death and
renewal processes and progress to chronic hepatitis,
fibrosis, cirrhosis and hepatocellular carcinoma
(Giannelli et al., 2003). Well, in experimental studies,
Paracetamol is widely used as a hepatotoxin in high dose
(overdose). The effectiveness of any preventive drug to
the liver relied mainly on its ability reducing injury
effects and the preservation of the natural function of the
liver, which is trouble by hepatotoxin (Dutta, et al.,
2013). However, the hepatotoxicity is caused by the
reaction metabolite of Paracetamoli.e N-acetyl-p-
benzoquinoneimine (NAPQI), which causes oxidative
stress and glutathione exhausting. (Boyd andBereczky.
1966). Paracetamol toxicity is due to the composing of
toxic metabolites when a part of it is metabolized by
cytochrome P-450. Introduction of cytochrome [Dahlin,
1984] or depletion of hepatic glutathione is a condition
A
B
C
D
Thuwaini et al. European Journal of Pharmaceutical and Medical Research
www.ejpmr.com
40
for paracetamol induced hepatotoxicity [Moron, 1979,
Gupt, 2006]. Nonetheless, the association of paracetamol
abusiveness and hepatotoxicity is getting significance
because of its usage as a suicidal agent either alone or in
combination with other analgesic mixtures. A dose of
15—50 gm/day is dangerous and may result in death of a
person due to hepatonecrosis (Prescott, et al., 1971).
Nonetheless, usually, AST and ALP are existing in high
concentration in liver. These enzymes are discharge from
the cells and their levels in the blood increment. Because
of hepatocyte necrosis or anomalies membrane
permeability (Shah, 2002).
However, this study appeared that the activities of serum
of AST and ALT are cytotolic indicator enzymes
inverting hepato-cellular necrosis, meaningly they are
discharged into the blood after cell membrane injuring.
Hence, we used the activities of AST, ALT and ALP in
the circulation as markers of hepatic damage.
However, Giving of clove extract at doses of 100 and
200 mg/kg, p.o., daily resulted in a significant sluggish
of moderate rise of serum enzyme markers in
paracetamol- induced group, in comparable to group II.
In which, proposing that the Clove extract as other plant
extracts may prevent hepatic injury associated with
overdose of PCM administration. Where, found there are
significant elevations in serum of (AST, ALT and ALP)
levels in PCM rats compared with healthy animals
(control group).This may relate to immoderate
accumulation of amino acids in the serum of animals as a
consequence of the moving of amino acids from protein
stores (Abolfathi, et al., 2012). These findings are
agreement with the results of a previous study in which
paracetamol was administered to rats (Sener, et al.,
2003). Furthermore, the bilirubin serum level took the
same manner. On the other word, a decline of serum
bilirubin in this study confirms the hepatoprotective
activity of clove extract. Conversely, the serum level of
hepatocellular albumin marker was significantly declined
in group II that treated with a paracetamol alone. That is
verifying the depleting of the liver function .in compared
to group I (control) (Table1). These scores were in line
with the results of previous studies that
weregivenparacetamol to rats (Sener, et al., 2003). It is
authenticated that the cloves extract to have a preventive
influence on plasma membrane of hepatocytes (Diego, et
al. 2014). On the other word, hepatic injury induced by
paracetamolin which caused a decrease in albumin. In
addition, hepatic transaminases such as AST and ALT It
is still considered the gleaming standards for assessing
the hepatic injury (Michaut,. et al., 2014). Moreover,
leakage of liver enzymes into blood when injury to the
liver due to increased permeability, injury and necrosis
of the hepatic cells.Therefore, AST, ALT and ALP levels
are elevating in liver toxicity (Dutta, et al., 2013).
Moreover, (Kumar et al., 2004), was appeared that is
One of the hallmark features of hepatic damage is
emergent leakage of cellular enzymes into plasma.
Besides, the range and sort of liver injury can be entered
based on the existence or absence of specific enzymes in
the blood surge. However, in this study, hepatoprotective
effect of clove plant extract is confirmed by the
amelioration ALT, AST and ALP levels. Where
simultaneously treatment with this extract inhibiting
PCM induced AST and ALT altitudes. Also emphasised
that there are Previous studies have documented
elevations of transaminases after APAP treatment
(Asha et al., 2004). The increment is time dependent
with importance elevation noted after two days
proposing severe hepatocellular injury caused by oozing
of these enzymes into circulation that is normally
cytoplasmic in position (Chung et al., 2001).
The biochemical results were also proved by histological
manifestations. The changes at most comprise
hepatocellular necrosis, fatty accumulation,
inflammatory cells infiltration and other histological
observations which were also in line with the scores of
other workers (Agbaje, et all., 2009;Matsura, et al.,
2006). However, histopathological outcomes further
added more verification to the hepatoprotective influence
of clove extract. Hence, the hepatoprotective impacts of
clove extract could be ascribed to its interference with
many pro- and inflammatory mediators which are getting
before hepatotoxicity.
CONCLUSION
The administration of cloves simultaneously with
paracetamol for end of experimental (4wks) led to
protects of hepatic cells by inhibition of oxidative effort
to paracetamol in rats. So it showed significantly
responding to changes in the functional test. These
results confirm paracetamol toxicity to the hepatic cells,
simultaneously has implications strong protection of
hepatic cells against hepatotoxicity in rats by its ability to
antioxidants. However, it is needed more study about the
mechanism by which clove extract makes its anti-
oxidants effects on human being which are still
unknowable. Though, considering our results in this
animal model, it is recommended that the clove plant
extract maybe used in the future as hepatoprotactive drug
from different liver injuries.
REFERENCES
1. Brown RA. Hepatic and renal damage with
paracetamoloverdosage. J Clin Pathol, 1968; 21(6):
793.
2. Karkosh AA, Study of in vitro antibacterial activity
of the essential oils of Cloves
(Syzygiumaromaticum) and the effect of
temperature on antibacterial activity. Euphrates
Journal of Agriculture Science, 2012; 4: 15-19.
3. Koba K,Nenonene AY, Raynaud C, Chaumont JP,
Sanda K, Antibacterial Activities of the Buds
Essential Oil of Syzygiumaromaticum (L.) 42
Merr.& Perry from Togo. Journal of Biologically
Active Products from Nature, 2011; 1: 42-51.
Thuwaini et al. European Journal of Pharmaceutical and Medical Research
www.ejpmr.com
41
4. Chaieb K, Zmantar T, Ksouri R, Hajlaoui H,
Mahdouani K, Abdelly C, Bakhrouf A. Antioxidant
properties of essential oil of Eugenia caryophyllata
and its antifungal activity against a large number of
clinical Candida species. Mycoses, 2007; 50: 403-
406.
5. Cortés-Rojas DF, De Souza CRF, Pereira Oliveira
W. Clove (Syzygiumaromaticum): a precious spice.
Asian Pacific Journal of Tropical Biomedicine,
2014; 4: 90-96.
6. Djilani A, Dicko A, The Therapeutic Benefits of
Essential Oils, Nutrition, Well-Being and Health. In:
Bouayed J (Ed.), 2012. ISBN: 978-953-51-0125-3,
In Tech, DOI: 10.5772/25344. Available from:
http://www.intechopen.com/books/nutrition-well-
being-and-health/the-therapeutic-benefits-of-
essential-oils access on 25 May, 2014.
7. Ibrahium MI, Abd El-Ghany ME, Ammar MS,
Effect of Clove Essential Oil as Antioxidant and
Antimicrobial Agent on Cake Shelf Life. World
Journal of Dairy & Food Sciences, 2013; 8: 140-
146.
8. Nosrati S, Esmailzadeh-Hosseini SA, Sarpeleh A,
Soflaei Shahrbabak M, SoflaeiShahrbabak Y,
Antifungal activity of spermint (Menthaspicata L.)
essential oil on Fusariumoxysporum f. sp. radicis-
cucumerinum the causal agent of stem and crown rot
of greenhouse cucumber in Yazd, Iran. In:
International Conference on Environmental and
Agricultural Engineering, Chengdu, China held on,
2011; 2011; 52-56.
9. di Paoli S, Giani TS, Presta, GA, Pereira MO, Da
Fonseca AD, Brandão-Neto J, Medeiros AD,
Santos-Filho SD, Bern ardo-Filho M, Effects of
Clove (Caryophyllusaromaticus L.) on the Labeling
of Blood Constituents with Technetium-99m and on
the Morphology of Red Blood Cells. Brazilian
Archives of Biology and Technologym, 2007; 50:
175-182.
10. Politeo O, Jukic M, Milos M, Comparison of
chemical composition and antioxidant activity of
glycosidically bound and free volatiles from clove
(Eugenia caryophyllata Thunb.). Journal of Food
Biochemistry, 2010; 34: 129-141.
11. Chomchalow, N., 1996. Spice production in Asia-an
overview. Proceeding of IBC’s Asia Spice Markets’
96 Conference. Singapore.
12. Mehana EE, Meki AR, Fazili KM. Ameliorated
effects of green tea extract on lead induced liver
toxicity in rats. ExpToxicolPathol, 2012; 64(4):
291–5.
13. Muriel P, Garciapina T, Perez-Alvarez V, Mourelle
M. Silymarin protects against paracetamol-induced
lipid peroxidation and liver damage. J Appl Toxicol,
1992; 12(6): 439–42.
14. Abolfathi Mohajeri D, Rezaie A, Nazeri M.
Protective effects of green tea extract against hepatic
tissue Injury in Streptozotocin-Induced diabetic rats.
Ecam, 2012; 2012: 10. AA,
15. Drotman R, Lawhorn G. Serum enzymes as
indicators of chemically induced liver damage. Drug
Chem Toxicol 1978; 1(2): 163–71.
16. Grover JK, Yadav S, Vats V. Medicinal plants of
India with antidiabetic potential. J Ethnopharmacol
2002; 81: 81–100.
17. Madkour FF, Abdel-Daim MM. Hepatoprotective
and antioxidant activity of dunaliellasalina in
paracetamol-induced acute toxicity in rats. Indian J
Pharm Sci., 2013; 75(6): 642-8.
18. Diego FR, Claudia RF, Wanderley PO. Clove
(Syzygiumaromaticum): a precious spice. Asian Pac
J Trop Biomed, 2014; 4(2): 90-6.
19. Kadarian C, Broussalis AM, Miño J, Lopez P,
Gorzalczany S, Ferraro G, et al. Hepatoprotective
activity of Achyroclinesatureioides (Lam) DC.
Pharm Res., 2002; 45(1): 57-61.
20. Dhinahar S, Lakshmi T. Role Of Botanicals As
Antimicrobial Agents In Management Of Dental
Infections – A Review. International Journal Of
Pharma And Bio Sciences, 2011; 2: 690-04.
21. Mroueh, M., Saab, Y., Rizkallah, R.,
Hepatoprotective activity of Centauriumerythraea on
acetaminophen-induced hepatotoxicity in rats.
Phytother. Res., 2004; 18, 431–433.
22. Hiraganahalli DB,Chandrasekaran CV, Dethe S,
Mundkinajeddu D, Pandre MK, Balachandran J, et
al. Hepatoprotective and antioxidant activity of
standardized herbal extracts. Phcog Mag., 2012; 8:
116–23.[PMC free article] [PubMed].
23. Heard K, Bui A, Mlynarchek SL, Green JL, Bond
GR, Clark RF, et al. Toxicity From Repeated Doses
of Acetaminophen in Children: Assessment of
Causality and Dose in Reported Cases. Am J Ther,
2012 Mar 8. [Medline]. [Full Text].
24. Basu S,Haldar N, Bhattacharya S, Biswas S, Biswas
M. Hepatoprotective activity of Litchi chinensis
leaves against paracetamolinduced liver damage in
rats. Middle East J Sci Res., 2014; 7(3): 292- 296.
25. Boyd, E.H. and G.M. Bereczky, Liver necrosis from
paracetamol. Br. J. Pharmacol., 1966; 26: 606-614.
26. Andrew, H.F., K.A. Jacobson, J. Rose and R. Zeller,
Hematoxylin and eosin staining of tissue and cell
sections. CSH Protoc, 2008. DOI: 10.1101/pdb.
prot4986.
27. Sabin, landan of Brian, and Everit, S. [Edit]. A
Handbook of statistical analyses lesing SPSS,
chapman of Hell CRC, was hington, 2004.
28. Giannelli G, Quaranta V, Antonaci S. Tissue
remodelling in liver diseases. HistolHistopathol,
2003; 18: 1267-74.
29. Shah M. Indian Drugs, 2002; 39: 333-337.
30. Dahlin D.Proc Natl Acad Sci., 1984; 81: 1327-1331.
15.
31. Moron MS, Depierre JW and Mannervik B.
BiochemBiophysActa. 1979; 582: 67-78. 16.
32. Gupta AK. J. Pharmacol. Toxicol. 2006; 1: 82-88.
33. Prescott, L.F., Wright, N., Roscoe, P. and Brown,
5.5. Plasmaparacetamol half-life and hepatic
Thuwaini et al. European Journal of Pharmaceutical and Medical Research
www.ejpmr.com
42
necrosis in patients with paracetamoloverdosage.
Lancet, 1971; 1: 519.
34. Sener, G., Sehirli, A.Ö., Ayanog˘lu-Dülger, G.,
Protective effects of melatonin, vitamin E and N-
acetylcysteine against acetaminophen toxicity in
mice. A comparative study. J. Pineal. Res., 2003; 35:
61–68.
35. Michaut, A., Moreau, C., Robin, M.-A., Fromenty,
B., 2014. Acetaminophen-induced liver injury in
obesity and nonalcoholic fatty liver disease. Liver
Int. http://dx.doi.org/10.1111/liv.12514.
36. Dutta BJ, Lahkar M, Augustine BB, Lihite RJ.
Hepatoprotective activity of Tamarind indica and
Homalomenaaromatica in rats. Int J Pharm Pharma
Sci., 2013; 5(2): 436-8.
37. Kumar, G., G.S. Banu, P.V. Pappa, M. Sundararajan
and M.R. Pandian, Hepatoprotective activity
of Trianthemaportulacastrum L. against paracetamol
and thioacetamide intoxication in albino rats. J.
Ethnopharmacol., 2004; 92: 37-40.
38. Asha, V.V., S. Akhila, P.J. Wills and A.
Subramoniam, Further studies on the antihepatotoxic
activity of Phyllanthusmaderaspatensis Linn. J.
Ethnopharmacol., 2004 92: 67-70.
39. Chung, Y.H., J.A. Kim, B.C. Song Song, I.H. Koh,
M.S. Lee, H.C. Eunsil, Y.S. Lee and D.J.
Su, Centrilobular hepatic necrosis; Isocitrate
dehydrogenase as a marker of centrilobular model of
rats. J. Gastroenterol. Hepatol, 2001; 16: 328-332.
40. Agbaje, E.O., A.A. Adeneye and A.O. Daramola,
Biochemical and toxicological studies of aqueous
extract of Syzigiumaromaticum (L.) Merry. and
Perry (Myrtaceae) in rodents. Afr. J. Tradit.
Complement Altern. Med., 2009; 6(3): 241-254.
41. Matsura T; Nishida T; Togawa A; Horie S; Ohata S;
Nakada J, Ishibe Y and Ohta Y. Mechanisms of
protection by melatonin against acetaminophen-
induced liver injury in mice. Journal of Pineal
Research, 2006; 41(3): 211-219.
