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Chronic Toxicity Studies of Aqueous Leaf Extract of Voacanga Africana in Wistar Rats * IGHODARO, IGBE

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Chronic Toxicity Studies of Aqueous Leaf Extract of Voacanga Africana in Wistar Rats * IGHODARO, IGBE JASEM ISSN 1119-8362 Full-text Available Online at J. Appl. Sci. Environ. Manage. Dec., 2015 JAllO UrightsRN AreservedL OF APPLIED S www.ajol.infoCIENCE A N D and EN VIRONVol.ME N19T (A4L) 6M39A -N 6A46G EMENT. www.bioline.org.br/ja Chronic Toxicity Studies of Aqueous Leaf Extract of Voacanga africana in Wistar Rats *1IGHODARO, IGBE; SILVANUS, INNIH2; STEPHEN, VINCENT OGEDENGBE1; LUCY, AMAMINA 2 Department of Pharmacology and Toxicology, Faculty of Pharmacy, University of Benin. 2Department of Anatomy, School of Basic Medical Sciences, University of Benin [email protected], [email protected], [email protected], [email protected] *Corresponding author: Ighodaro Igbe Department of Pharmacology & Toxicology, Faculty of Pharmacy, University of Benin, Benin City, Nigeria. PMB1154 [email protected]. +2348166058559 ABSTRACT: Voacanga Africana stapf (Apocynaceae) leaves is being used in herbal medicine to treat leprosy, diarrhoea, generalized oedema and convulsion in children as an infant tonic and the present study was aimed at its toxicological evaluation in wistar rats. The sub-acute toxicity was evaluated after administering daily oral doses of Voacanga Africana stapf (100, 400 and 800 mg/kg) for 28 days after which the effect on anthropometric, haematological and histopathological parameters were assessed. There was a significant reduction (p<0.05) in the pattern of weight gain in the female albino rats and alkaline phosphatase (ALP) but no significant difference in the organ weight index in all selected organs. There were no gross abnormalities or histopathological changes observed among any the groups treated. The results suggest that the aqueous leaf extract of Voacanga africana can be considered relatively safe on chronic administration to rats and may cause reduction in weight gain in female rats probably due to changes in female hormones. © JASEM http://dx.doi.org/10.4314/jasem.v19i4.11 KEYWORDS; Voacanga africana, toxicity, weight gain, haematology INTRODUCTION (Kokiyar.) It is a perennial plant grows to the As the use of medicinal plants increases, height of 6m. The flower is bisexual, the leaves are experimental screening of their toxicity is crucial to broad and oval and up to 30cm long (Iwu, 1993). guarantee the safety of users (Asgarpanah and The milky latex of the plant is applied to wounds in Ramezanloo, 2012). Toxicity is an expression of Nigeria and Senegal. Tea made from the leaves being poisonous, indicating the state of adverse serves as a strengthen portion that relieves fatigue effects caused by the interaction between toxicants and shortness of breath. It is also used to prevent and cells (Syahmi et al, 2010). Natural products are premature childbirth and to treat painful the cornerstone of health care delivery especially in menstruation (Sofowora, 1993). In Senegal, a leaf resource poor settings. Present estimates indicate decoction is drunk as a tonic and against fatigue. In that about eighty percent of the world’s population Cote d’ivoire a decoction of the leaves is applied as relies on traditional medicine for health care a wash against oedema and is used as a friction and delivery (Peter, 1995). So much has been done in is drunk in the treatment of leprosy. The pulp from screening medicinal plants for efficacy based on the leaves or stem bark is applied to soothe traditional claims while less emphasis is placed on convulsions in children and the juice is put in the the issue of safety, as reports of efficacy far nostrils as a tranquillizer (Burkill et al, 1985). The outnumber those of toxicity, probably as a result of dried and powdered roots without the outer bark the greater demands for resources and time such are mixed with porridge and taken against kidney exercise warrant. Pharmacological and toxicological troubles and menstruations problems in women. evaluations of medicinal plants are essential for drug The wood is used to make musical instruments and development (Sofowora, 1993) also used for firewood. Good fiber can be obtained from the bark and is made into rope. (Tona et al, Voacanga africana is a small tropical tree, 1998). In the present study, the subchronic toxicity distributed mainly in West Africa. It is locally of the aqueous extract of the leaves was evaluated known by different names such as; in Igbo to assess its safety, since the findings are important (petepete), in Yoruba (Ako-Dodo) and in Hausa Corresponding author Email: [email protected] Chronic Toxicity Studies of Aqueous Leaf Extract of Voacanga Africana 640 considering the usage of the plants by human stored in 20ml sample bottles (Axiom, Zhanjiang beings. Gong Jong medical technology Co. Ltd, China) containing 10% formyl saline. Thereafter, they were MATERIALS AND METHODS processed for histopathological studies. Plant Materials and Extraction: The leaves of Voacanga africana were collected from Okhoro Effect of Voacanga africana on body weight.: In the village in Benin City, Nigeria in the month of course of the 28-day oral treatment, body weights of March, 2015. The plant was identified and rats were regularly taken at a 7day interval. Weight authenticated by Prof. Macdonald Idu of the changes were calculated in respect of the initial Department of Plant Biology and Biotechnology, body weights on day zero (0). University of Benin, Benin City were a voucher specimen was deposited. The leaves were sun dried Effect of Voacanga africana on organ weight index: to a constant weight over a 14 day period. The dried The organs (heart, lungs, liver, spleen and kidney) leaves were then powdered using a mechanical were carefully dissected out and freed from grinder. The powdered material (500g) of Voacanga adjoining supporting connective tissues. The organs africana was boiled in 2.5L of distilled water for 30 were gently rinsed in normal saline, blotted with minutes, and allowed to cool and then filtered. The filter paper and weighed. The organ index was then filtrate was then concentrated in a rotar vapor to calculated using the formula: give a total yield of 13.3% (w/w). The dried extract Organ weight index= was preserved in an air tight clean glass container and kept in a refrigerator maintained at -4°C until Haematological assay: The blood samples collected use. into the lithium heparinized samples bottles were for determination of the red cell count (RBC), packed Experimental Animals: Experiments were cell volume (PCV), haemoglobin concentration performed using albino wistar rats of either sex (Hb), and platelet count (PLT), total white blood (180-220g). The animals were obtained from cell counts and its differentials using Automated Animal House, School of Basic medical Science, Haematology System. (Diatron Abacus junior Department of Anatomy, University of Benin, Benin hematology analyzer). City, Nigeria. The animals were acclimatized for two weeks and fed with standard feed (from Ewu Biochemical assay: Biochemical analysis was feed and flours mills Ltd, Ewu, Edo state, Nigeria) performed on serum obtained after centrifugation of and tap water ad libitum. Animals were exposed to white blood (without Anticoagulant) at 2500rpm for natural lighting conditions and were handled in 5 minutes. Standardized diagnostic kits ( Randox by accordance with international principles guiding the Randox laboratories LTD., United Kingdom). use and handling of experimental animals (National Determination of the following biochemical Institutes for Health, USA, 2002) approved by the parameters: alanine aminotransferase (ALT), Faculty of Pharmacy Ethics Committee on Animals. aspartate aminotransferase (AST), creatinine, alkaline phosphatase, urea, bicarbonate, total proteins, albumin, total and conjugated bilirubin. Toxicity Study: Animals were randomly allotted to 4 groups of 10 rats per group containing five males, Serum ALT, AST and ALP were measured using five females. Animals received distilled water in the enzyme kinetic method of Reitan and Franklin group I while groups II to IV received 100mg/kg, (1957). Urea was measured using urease-Berthelot 400mg/kg and 800mg/kg of the extract respectively, method (Fawcett and Scott,1960). Estimation of The extract was orally administered daily at single creatinine was done by the Jaffe’s reaction method doses for 28 days and the rats were closely observed (Biod and Sirota, 1948). The total protein was for the general and behavioral signs of toxicity, estimated by Biuret method (Tietz, 1970) while that body, weight changes and mortality during the of albumin was determined by Bromocresol green entire period of the experiment. At the end of the (Lowry et al, 1957). The total bilirubin and the 28-day treatment period, animals were sacrificed conjugated bilirubin were determined by Jendrassik- under chloroform anaesthesia. Blood samples were Grof method (Spencer and Price, 1977). withdrawn by cardiac puncture into plain or lithium heparinized sample bottles. Histopathology: Spleen, liver, lungs, heart and Selected internal organs such as liver, heart, spleen, kidney were fixed immediately in 10% formalsaline kidney and lungs were collected, weighed and for routine histopathological examination. The *1IGHODARO, IGBE; SILVANUS, INNIH2; STEPHEN, VINCENT OGEDENGBE1; LUCY, AMAMINA 2 Chronic Toxicity Studies of Aqueous Leaf Extract of Voacanga Africana 641 tissues were embedded in molten paraffin wax and Statistical analysis Data were expressed as then sectioned, stained with haematoxylin and eosin mean±SEM. The data were analyzed using one and were examined under
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