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311
A HISTOLOGICAL STUDIES OF THE AMELIORATIVE EFFECT OF
TURMERIC
(CURCUMA LONGA)
ON THE LIVER OF
METRONIDAZOLE-TREATED ALBINO WISTAR RATS
Elioku.O.Grace1, Anyabolu A.E1, Ofoego U.C.1, *1Okafor I. A.
1
Department of Anatomy, Faculty of Basic Medical Sciences, College of Health Sciences,
Nnamdi Azikiwe University. Nnewi Campus, Anambra State Nigeria.
ABSTRACT
This study was carried out to evaluate the ameliorative effect of
turmeric on the cytoarchitecture of the liver of metronidazole (MTZ) -
treated rats. Twenty adult wistar rats weighing 140-200 gramms were
used. The animals were randomly divided into four groups of five (5)
each. The groups were labelled A,B,C and D. Group A animals served
as the control and received feed and water only. Group B received low
dose of metronidazole (200mg/kg/day), Group C received high dose of
metronidazole (400mg/kg/day), Group D received high dose of
metronidazole (400mg/kg/day) + Turmeric (400mg/kg/day). The
administration lasted for twenty eight (28) days and the animals were
weighed. At the end of experimental period, the animals were
sulphocated via Diethylether and dissected. The liver were weighed
and trimmed down for histological studies. The final body weight for
group C treated with high dose of metronidazole (400mg/kg) was significantly lower
(p>0.05) than group (B and D); but significantly higher (p>0.05) than the group A (control).
The final body weight for group D treated with metronidazole+turmeric was significantly
higher (P>0.05) than the control and other experimental groups (B and C) animals. The
relative liver weight for group D (400mg/kg/day Metronidazole+Turmeric administered)
were relatively higher than that of group A (control) and other experimental groups (B and
C). Histological observation showed the presence of prominent necrosis and evidence of
cellular infilteration into stroma in group C (400mg/kg metronidazole). While group D
treated with turmeric showed mild vacuolation and mild necrosis and evidence of slight
Volume 4, Issue 9, 311-320. Research Article ISSN 2277– 7105
Article Received on 30 June 2015,
Revised on 24 July 2015, Accepted on 19 Aug 2015
*Correspondence for Author
Okafor I. A.
Department of Anatomy,
Faculty of Basic Medical
Sciences, College of
Health Sciences, Nnamdi
Azikiwe University.
Nnewi Campus, Anambra
State Nigeria.
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312
hemorrhage. It can be concluded that metronidazole (MTZ) can cause toxicity, and turmeric
can provide ameliorative effect against metronidazole toxicity.
KEYWORDS: Metronidazole, Turmeric, Ameliorate, Histology, liver, Albino Wistar Rats.
1. INTRODUCTION
Metronidazole (MTZ) is a 5-nitroimidazole drug widely used in veterinary and human
medicine for the treatment of trichomoniasis, giardiasis, amoebiasis and anaerobic bacterial
infection.[1,2,3] It also shares the trade name flagyl or Nidagyl. Metronidnzole is metabolized
in the liver and excreted mainly via the kidney in urine and to a lesser extent through the
intestinal wall with feaces.[4] The drug is available in various formulations like tablet (200mg,
250mg, 400mg, and 500mg) hard capsule (500mg); oral suspension (40mg/ml); Suppositories
(500mg) and solution for injection (5mg/ml.[5]
Metronidazole has been listed by United States national toxicology program (NTP) as
reasonably anticipated to be a human carcinogen according to WHO International Agency for
Research on Cancer (IAR).[6,7]
Animal studies on oral metronidazole has shown increased incidence of tumor in the lungs,
liver, testes reticulum, mammary gland and pituitary gland in certain rodent species.[8,9] Other
studies also shows that high dose of metronidazole causes hepatotoxicity in rats.[10]
Turmeric is a perennial herb of ginger family (Ginger baracae), which is also known as
Curcuma longa.. It is a traditional curry spice used as a yellow food colouring. Turmeric has
been used in traditional medicine in India and ancient Egypt for at least 6000 years and has
been extensively cultivated in tropical areas of Asia, and to a lesser extent in Africa. In
approximately the past 50 years, turmeric has been subjected to numerous trials and studies
and it has been validated and clarified by modern science.[11,12] The most important
component of turmeric is curcumin. Curcumin has been found to have hepatoprotective
characteristics. Animal studies has demonstrated it hepatoprotective effect to from a variety
of hepatotoxic insults like ethanol, paracetamol, metronidazole etc.[13,14,15]
The liver is the work horse of the human body and the most complex organ involved in
metabolism of food and drugs.[12] The liver is the major target organ for chemically induced
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313
approved drugs from the United States market. This also accounts for more than 50% of cases
of liver failure in the united state today.[16]
Liver disease is one of the major causes of the mortality in man and animals all over the
globe, with hepatotoxicity being a major contributing factor.[17] The liver filters toxic
substances like alcohol, chemotherapeutic drugs, antibiotics and toxicants from the body. If
accumulation of these toxins is faster than the liver metabolizing ability, hepatic damage may
occur.[18]
2. MATERIALS AND METHOD
2.1 BREEDING OF ANIMALS
Tweenty Adult Wistar Rats were purchased from a local farm at Nsukka in Enugu State,
Nigeria. They were breed in Experimental Animal House of the Department of Anatomy,
Nnamdi Azikiwe University, College of health science, Nnewi Campus. Anambra state
Nigeria. They were allowed for a period of two weeks acclimatization under normal
temperature (27ºc – 30ºc) before their weight were taken. They were feed with Growers
Mash, product of Premier Feed Mills Company Limited in Sapele, Delta State Nigeria.
2.2 DRUG PREPARATION AND COLLECTION
The drug used for this research work includes the following as stated below: Metronidazole
by May & Baker Nig. Plc; 400mg and 200mg with expiration date of 2018, were purchased at
Gods Will Pharmacy Nnewi Anambra State Nigeria. Metronidazole was grounded to fine
powder and dissolved in water (50mls of water for 800g of metronidazole) for 10min before
administration daily to allow for proper dissolution. Turmeric powder with expiration date of
2016 was purchased from the Main Market at Nnewi Town, Anambra State. Turmeric was
made and packaged in India by TRS ASIAS FINEST FOOD, INDIA. Before administration,
the turmeric powder was weighed and dissolved in water (2g dissolved in 10mls of water).
Turmeric was administered in solution form. Weighing of extract was done weekly with an
Electronic Weighing Balance with accuracy range of 100g
2.3 EXPERIMENTAL PROTOCOL (DESIGN AND PROCEEDURE)
The Twenty Adult Male wistar Rats were weighed and allocated into four groups of five
animals each. The groups were designated as groups A,B,C and D. Group A animals served
as control and received water and feed only. The experimental groups B,C and D received
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314
metronidazole; Group C received 400mg/kg bodyweight/day metronidazole; Group D
received 400mg/kg/bodyweight/day metronidazole + 400mg/kg body weight Turmeric. The
drugs were administered once a day between the hour of 7:am- 12:pm; for period of
tweenty-eight days. The drug were administered orally using syringes (5mls & 2.5mls). After the
twenty-eight day, the animals were weighed and their weight recorded. Tweenty-four Hours
after the last administration, the animals were sulphocated via Diethylether and were
dissected. Liver tissue were removed from the animals and weighed. They were trimmed
down to a size of 3mm x 3mm thick and fixed in Zenkers fluid for four (4) hours for
histological studies. The tissue slide were prepared in the Histopathological Department of
the Nnamdi Azikiwe University Teaching Hospital Nnewi Anambra State.
2.4 STATITICAL ANALYSIS
The data are reported as mean and standard deviations. Statitical comparison between group
mean were done by one-way analysis of variance. P value less than (<) 0.05 were considered
significant.
3. RESULTS
[image:4.595.67.561.470.766.2]3.1 MORPHOMETRIC ANALYSIS OF BODY WEIGHTS
Table 4.2: Comparison of mean initial and final body weight and weight change in all the groups (A,B,C and D)
(Mean ± SD given for each measurement)
GRP A GRP B GRP C GRP D t - MEAN PROB OF SIG
Initiai Bwt 164.00±13.41 180.00±15.81 180.00±15.81 176.00±8.94 30.56 >0.05
Final Bwt 190.00±14.14 210.00±15.81 208.00±8.36 212.00±8.36 43.00 <0.05
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[image:5.595.143.453.283.506.2]315 MORPHOMETRIC ANALYSIS OF LIVER WEIGHTS
Table 4:3: comparison of mean relative liver weight for group A (control) and Experimental groups (B, C and D)
(Mean ± SD given for each measurement)
The bar chart representation of the relative liver weight of the various groups. The group D
(400mg/kg metronidazole+400mg/kg turmeric administered) were significantly higher
(p>0.05) than the control group (A) and groups B and C as shown in 4.3.1
MICROGRAPH 1 (Group A; control): The micrograph shows normal hepatic architecture.
GRP A GRP B GRP C GRP D t - MEAN PROB OF SIG
Weight of
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316 MICROGRAPH 2 (Treated with 200mg metronidazole; Group B): The micrograph shows mild infiltration of the lymphoplasmaticic cells intralobularly stained by H&E technique ×100.
MICROGRAPH 3 (Treated with 400mg Metronizdazole; Group C): The micrograph shows disruption of liver cells. It shows prominent necrosis and there is evidence of cellular infiltration into stroma.
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317 DISCUSSION
Metronidazole has been listed by United States national toxicology program (NTP) as
reasonably anticipated to be a human carcinogen according to WHO International Agency for
Research on Cancer (IARC).[6,7]
Animal studies on oral metronidazole has shown increased incidence of tumor in the lungs,
liver, testes reticulum, mammary gland and pituitary gland in certain rodent species.[8,9] Other
studies also shows that high dose of metronidazole cause hepatotoxicity in rats.[10] The results
of this work agrees with previous researches that metronidazole has toxicological effect on
the liver of wistar rats. In the studies by sama 2012, it was revealed that the liver showed
minute foci of hepatic cell necrosis and associated mononuclear cell infiltrate were evident
almost adjacent to the portal areas.
Turmeric (Curcuma longa) is one of the most useful herbal medicinal plants. Extensive
researches have proven that most of the turmeric hepatoprotective activities are due to
curcumin. Histopathological studies has also shown that administration of aqueous extract of
turmeric offered significant protection from the damaging actions of ethanol induced
hepatotoxicity in albino rats.[13] Animal studies has demonstrated it hepatoprotective effect
from a variety of hepatotoxic insults like paracetamol, metronidazole etc.[14,15] The result of
this present study has shown that turmeric was able to ameliorate the toxic effect of
metronidazole on the liver of the group treated with turmeric when compared with those
treated with high dose of metronidazole and this correlates with the report by Ishwin sigh et al,(2012). And also shows similarity with the report by Bruk et al (2007).[19] who prostulated that curcumin was able to inhibit the development of Thioacetamide (TAA) induced liver
cirrhosis.
It was observed that the histology of the liver of control group showed normal architecture.
The group treated with of MTZ 200mg/kg/day (low dose) showed evidence of mild
infiltration of lymphoplasmaticic cells, and this corresponds with the report by Sama, (2012).
Those treated with MTZ 400mg/kg/day (high dose) showed prominent necrosis and evidence
of cellular infiltration. And this agrees with the report by Sama, (2012).The last group treated
with with MTZ 400kg/mg/day + Turmeric 400mg/kg/day. Showed mild necrosis and mild
vacoulation and evidence of slight haemorrhage and this result is similar to the prostulated by
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Observation of the body weight difference in groups reveals gradual increase in weight of
animals for the control group A. This could have been physiological as the only substance
they were exposed to was water and feed. Comparing the results of weight difference reveals
significant decrease in weight (p<0.05) by the group treated with high dose MTZ when
compared with the other treatment groups. This is probably as a result of loss of appetite by
the animals in the group. The group that was treated with metronidazole + turmeric showed
significant increase in weight when compared with the control group and other treatment
groups. Turmeric in this context functioned primarily as a dietary supplement enhancing
growth.
The relative organ weights also showed significant differences in groups. There was relative
increase in liver weight for turmeric + metronidazole treated animals when compared with
the control and metronidazole alone treated animals (low and high dosage). This organ
weight increase was due to the nutritional and protective value added by turmeric powder.
There was also relative increase in the liver weight of animals treated with high dose of
metronidazole when compared with the control group and low dose metronidazole treated
group. This organ weight increase was irrespective of the fact that there was a relative body
weight loss in this group. This could have been pathological and one may deduce that the
increase in liver weight was not growth but inflammation. Antioxidant properties of Turmeric
could have been responsible for the control or prevention of inflammation in the group
treated with it.
Administration of low dose of metronidazole alone did not cause weight loss to the animals
compared with the animals in control group and those treated with high dose of
metronidazole. By these observation one may deduce that administration of Turmeric may
boost the tolerance capacity for metronidazole induced toxicity.
Thus, the protective and ameliorative effect of Turmeric against metronidazole dosage liver
damage recorded in the present study is attributed to their antioxidant properties.
CONCLUSION
Turmeric was able to ameliorate the change in the histoarchitecture of liver tissues of the rats.
This study has demonstrated the potential ability of turmeric to protect against the harmful
effect of metronidazole in the liver of rats. Rats tissues are very similar in many aspects to
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319
humans for a long period of time, can cause toxicity on the liver; and if turmeric is
administered to individuals exposed to high dosage metronidazole toxicity, it could provide
some protection against metronidazole toxicity and perhaps ameliorate the effects of
metronidazole toxicity on the liver. This study has opened a way for possible means of
preventing damage to man’s tissues by metronidazole. However, further work is needed
particularly on the effect of this drug on other organs like kidney, spleen, lungs etc.
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