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Attenuation of chloroquine-induced hepatoxicity and renal damage by Gnetum bucholzianum leaf extract.

Gnetum bucholzianum belongs to the gnetaceae family usually with climbing jointed stems (1). It is a small tree with tiered branches and divaricate branchlets having broad glossy dark green leaves. It is found in Amucha Njaba LGA, Imo State Nigeria as well as other parts of the South Eastern Nigeria. In Igbo land it is called "Ukazi" or "Ukasi" while Efik call it afang. Nutritionally G. bucholzianum is very rich in proteins and minerals. The leaves contain high nutritional values as it contains eight amino acids in significant quantities (2). Dishes based on G. bucholzianum leaves are prominent on the menu list in some restaurants and hotels in Owerri, Nigeria.

Chloroquine is a member of an important series of chemically related anti-malarial agents, the quinolone derivatives. It is a synthetic drug used in the treatment of malaria. Being a 4-aminoquinoline, it is a rapidly acting blood schizontocide with some gametocytocidal activity (2). In this study, the effect of G. bucholzianum was evaluated to provide information on its attenuation effect on chloroquine-induced hepatoxicity and renal damage in Wistar rats.

Chloroquine (Emzor) was purchased from a standard pharmacy shop in Owerri, Imo State, Nigeria. The tablets were dissolved in distilled water according to the required concentrations required for administration to Wistar rats on the basis of their body weight.

G. bucholzianum was obtained from the Ekeonunwa market in Owerri Nigeria. The botanical identification and authentication was confirmed by Dr. C. Okere (Head of Department of Plant Science and Biotechnology, Imo State University ,Owerri). The plant material was sun dried for seven days. The dried leaves of the G. bucholzianum were milled to achieve a coarse powder used for extraction. The powder was macerated in a 400g percolator with 200ml of distilled water. The mixture was allowed to stand for 48 hours after which it was filtered. The filtrate was then placed in an oven to evaporate and the solid residue referred to as extract. The appropriate concentrations of the extract were made in distilled water for the experiments. Hence, the following concentrations: 200mg and 400mg were prepared.

Wistar albino rats, weighing between 160 and 240g and aged 8-12 weeks, were used in the study. These animals were obtained from the Animal House of College of Medicine and Health Sciences, Imo State University, Owerri Nigeria. They were kept under standard laboratory conditions, fed with commercial growers mash (Tops Feeds Ltd, Sapele, Nigeria). Water and feed were provided ad libitum. The animals were left for two weeks to acclimatize. The experimental protocol was approved by the local ethical committee for animal experimentation. The animals were handled in accordance with institutional guidelines for the care and use of animals for experimental purposes.

The animals were randomly assigned to four experimental groups (n = 6 in each group). The first group of animals, which served as the control group, was given distilled water. Groups II, III and 1V were given chloroquine (970mg/kg body weight) and G. bucholzianum extract (200mg/body weight); chloroquine and G. bucholzianum extract (400mg/body weight) respectively for 14 days. In all groups the drug was administered through the oral route using a feeding tube attached to a 5ml syringe. All animals were allowed free access to food and water throughout the experiment.

Twenty four hours after the last doses were administered, the animals were sacrificed and blood collected for biochemical analysis.

In this study, chloroquine administration in a dose of 970 mg body weight of Wistar rats elevated serum hepatic and renal parameters. The damage to the liver and kidney resulted in significantly increased bilirubin, AST, ALT, ALP, urea and creatinine levels.

The liver and renal damage may be due to generation of free radicals by chloroquine overdose which is also partly responsible for its anti-malaria effects (4). This harmful effect could be caused by free radicals produced during peroxide formation. The level of hydroxylchoroquine treatment may be responsible for the hepatic and renal impairment (5). However, the simultaneous administration of Gnetum bucholzianum significantly reduced the effect of chloroquine by attenuating hepatic and renal parameters. This could be of importance in Nigeria if Gnetum bucholzianum is consumed while on chloroquine medication for malaria.


(1.) Nkwatoh AF, Labode P, Iyassa SM, Nkwatoh FW, Ndumbe NL, Ewane ME. Harvesting and marketing of Gnetum species (Engl) in Cameroon and Nigeria. J Ecol Nat Environ 2010; 2: 187-193.

(2.) Lowe J. Gnetum in west Africa. Nigeria Field 1984; 49 (1-4): 99-104.

(3.) Sowunmi A, Fehintola FA, Adedeji AA, Falade AG, Falade CO, Akinyinka OO, Oduola AMJ. Comparative efficacy of chloroquine plus chlorpheniramine alone and in a sequential combination with sulfadoxine-pyrimethamine for the treatment of acute, uncomplicated, falciparum malaria in children. Ann Trop Med Parasitol 2000; 94: 209-217.

(4.) Ansari NM, Houlihan L, Hussain B, Pieroni A. Antioxidant activity of five vegetables traditionally consumed by south-Asian migrants in Bradford, Yorkshire, UK. Phytother Res 2005; 19: 907-911.

(5.) Nnodim JK, Adamma E, Austin A, Chukwunyere NNE.Alterations in biochemical parameters of wistar rats administrated with sulfudoxine and pyrimethamine (FansidarR). Al Almeen J Med Sci 2010; 3: 317-321.

Author information

Johnkennedy Nnodim *, BMLS MSc AMLSCN, Lecturer [1] Augustine Ihim, BMLS MSc AMLSCN, Medical Laboratory Scientist [2] Hellen Ifeoma Udujih, BMLS MSc AMLSCN, Lecturer [1]

[1] Department of Medical Laboratory Science, Faculty of Health Science, Imo State University, Imo State, Nigeria [2] Department of Medical Laboratory Science, Nnamdi Azikiwe University, Nnewi Campus, Anambra State, Nigeria * To whom correspondence should be addressed. Email:
Table 1. Hepatic and renal function parameters in rats given
chloroquine with 200mg/kg and 400mg/kg body weight of extract
of G. bucholzianum.

Group Treatment AST IU/L ALT IU/L

1 Control 14.3 [+ or -] 2.3 12.1 [+ or -] 2.8
2 Chloroquine 29.5 [+ or -] 3.6 * 23.3 [+ or -] 2.7 *
 Chloroquine 20.0 [+ or -] 1.8 * 17.5 [+ or -] 2.9 *
3 +200 Gb exract
4 Chloroquine 17.3 [+ or -] 3.1 * 15.9 [+ or -] 3.5 *
 +400 Gb extract

Group Treatment ALP IU/L Bilirubin mg/dl

1 Control 62.7 [+ or -] 8.4 0.5 [+ or -] 0.2
2 Chloroquine 89.4 [+ or -] 9.1 * 1.6 [+ or -] 0.99 *
 Chloroquine 69.6 [+ or -] 9.4 * 1.0 [+ or -] 0.08 *
3 +200 Gb exract
4 Chloroquine 66.3 [+ or -] 7.8 * 0.8 [+ or -] 0.19 *
 +400 Gb extract

Group Treatment Urea Creatinine
 mg/dl mg/dl
1 Control 24.7 [+ or -] 3.5 0.7 [+ or -] 0.11
2 Chloroquine 57.3 [+ or -] 5.9 * 2.2 [+ or -] 0.09 *
 Chloroquine 36.4 [+ or -] 4.7 * 1.7 [+ or -] 0.13 *
3 +200 Gb exract
4 Chloroquine 31.7 [+ or -] 5.2 * 1.3 [+ or -] 0.15 *
 +400 Gb extract

Gb: G. bucholzianum
* Significantly different from control (P<0.05)
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Author:Nnodim, Johnkennedy; Ihim, Augustine; Udujih, Hellen Ifeoma
Publication:New Zealand Journal of Medical Laboratory Science
Date:Aug 1, 2012
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