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A HISTOLOGICAL STUDIES OF THE AMELIORATIVE EFFECT OF TURMERIC (CURCUMA LONGA) ON THE LIVER OF METRONIDAZOLE TREATED ALBINO WISTAR RATS

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www.wjpr.net Vol 4, Issue 09, 2015.

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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www.wjpr.net Vol 4, Issue 09, 2015.

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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www.wjpr.net Vol 4, Issue 09, 2015.

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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318

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.

REFEREENCES

1. Reynold A.V., J.M.T. Hamilton-Miller and W. Brumfill A comparison of the invitro

activity of metronidazole, tinidazole and nimorazole against Gram –ve. Anaerobic

bacilli. Journal of clinical pathology., 1975; 28: 775-778.

2. Bergan, T., Antibacterial activity and pharmacokinetics of nitroimidazole. A review.

Scandivananian Journal of infectious Diseases., 1985; 46: 64-71

3. Frey. H.H and W. Loscher Lehrbuchder pharmakologie und Toxicologie fur die

Veterinar Medizin. Enke Stuttgant, 502-503. International Journal of Pharmacology.,

1996; (3): 252-256. Asian Network of Scientific Information.

4. Finch, R.G. and I.S. Snyder, (1986); Antiprotozoan drugs. In: Modern

Pharmacology.Eds.: C.R. Craige and R.E. Stitzel. Little, Brown co. Boston, pp:729-740

5. Rapporteur’s Public Assessment Report For Pediatrics Studies, (2010); Submitted in

accordance with Article45 of Regulation (EC) No 1901/2006

6. Report on carcinogens, Twelft Edition (2011); U.S. Department of health and human

services,Public Health Serrvices, National toxicology program.

7. International agency for research on cancer (IARC) (May 2010); Agents classified by the

IARC Monographs. World Health Organization.

8. Bendesky, A; Menwndez, D; Ostrosky-Wegman, p “Is Metronidazole Carcinogenicc?.

Mutation Research., 2002; 511(2): 133-44.

9. New Zealand Data Sheet, (2013); Metronidazole Description Pharmacology. MedSafe.

Hppt:/www.medsafe.govt.nz/profs/datasheet.

10. Samah S. Oda (2012); Histopathological and Alterations of Metrondazole- Induced

Toxicity in male Rats. Global veterinaria., 2012; 9(3): 303-310. ISSN1992-6172.

11.Planet Curcumin, (2010); Curcumin for linel/ Antimicrobial Activity.

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www.wjpr.net Vol 4, Issue 09, 2015.

320

13. Ishwin Sigh, S. Vetriselvan, J. Shankar, S. Gayathini, G. Shereenjeet, A. Yaashini

(2012); Hepatoprotective activity of aqueous extract of Curcuma longa in ethanol induced

hepatotoxicity in Albino wistar rats. International Journal Of Pharmacology.

14.Somchit M.N., A. Zurani, A. Ahmad Bustaman, M.R Sulaiman and R. Noratulina,

(2005); Protective Activity of Turmeric (Curcuma longa) in Paracetamol-Induced Hepatotoxicity in Rats.

15.Ali Noorafshan, Elham Aliabadi and Hossein Mirkhani, Ameliorative effect of Curcumin

on the Seminiferous Epithelium in Metronidazole - Treated Mice: A Sterological Study.

Toxicol pathology., 2010; 38: 366.

16. Lee WM, (2004); Drug- induced hepatotoxicity. New Engl. J. Med 349: 474-485

Liver-wikipedia. en.m.Liver-wikipedia.org/wiki/liver. Last modified on 18th june 2014.

17. Nadeem M.P.C, Dandiya P.C, K.V Pasha, Pasha M, Imran D, Balani K, Vohora S.B

Hepatoprotective acivity of solanum nigrum fruit. Fitotorpia., 1997; 68: 245-251.

18. Bigoniyan P, Singh C.S, Shukla A, A comprehensive review of different liver toxicants

used in experiment and pharmacology. IJPSDR., 2009; 1(3): 124-135.

19.Bruk Rafael, Ashkenazi Michal, Goldiner Ilana, Shapiro Haim, Aeed Hussein, Genina

Olga, Helpern Zamir, Pines Mark, (2007); Prevention of liver cirrhosis in rats by

curcumin. Liver International: Official Journal of the International Association for the

Figure

Table 4.2: Comparison of mean initial and final body weight and weight change in all
Table 4:3: comparison of mean relative liver weight for group A (control) and

References

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