You are on page 1of 5

Continental J.

Pharmaceutical Sciences 4: 35 - 39, 2010 © Wilolud Journals, 2010

ISSN: 2141 - 4149 http://www.wiloludjournal.com

TOXICITY AND HAEMATOLOGY STUDIES OF THE ROOT BARK OF HYMENOCARDIA ACIDA, TUL (EUPHHORBIACEAE) P.N. Olotu1, H. Ibrahim2, N Iliyas2 and J.S Gushit3 Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, University of Jos, Jos, Nigeria. 2 Department of Pharmacognosy and Drug Development, Faculty of Pharmaceutical Sciences, Ahmadu Bello University Zaria, Nigeria. 3 Department of Science Laboratory Technology, Faculty of Natural Sciences University of Jos, Nigeria.
1

ABSTRACT The research work covers the toxicities and haematology studies of the root bark of Hymenocardia acida which is claimed by the Hausa in the Northern Nigeria to be used traditionally for the treatment of headache, chest-pain, rheumatic pain, toothache, ear pain, migraine and sickle cell crisis. The plant was safe orally on acute toxicity investigation on mice. There was no observable LD50. The mice survived even the doses greater than 5000mg/kg and up to 11000mg/kg. The histopathological studies on chronic administration of the root bark in mice fed with 25% w/w of amended diet showed no observable organ damage but the mice fed with 50%w/w amended diet showed a focal area of hepatic necrosis and the globule cells of the intestine were covered with progressive mucin and lymphocyte proliferations were observed with the spleen. This means that the long term use of the plant in traditional practice should be cautioned. Furthermore, the body weight, feed and water-intake increase with the mice fed with the lower concentration of the amended diet (25% w/w) but decreased with the higher concentration (50% w/w) at P<0.05. There was also a prominent change with the post treated mice. This result may suggest the use of the plant for nutritional purposes at lower concentration. Analysis of the blood parameters showed very significant increase with the RBC. This justifies the claim of the plant in the treatment of anaemia. Analysis of the plant on the remaining blood parameters was not significant. KEYWORDS: Acute Euphorbiaceae. toxicity, Chronic toxicity, Haematology, Hymenocardia acida and

INTRODUCTION The plant Hymenocardia acida is a tree of about 6 m high, gnarled and twisted with characteristics rough rustyred bark, of the wooded savanna throughout the region from Senegal to west Cameroons, and widespread in tropical Africa. The wood is light brown or pink, darkening to orange, close- grained, with conspicuous annual rings, and hard. It has been said to be brittle and good only for firewood. This may be reflected in certain Ivorian names meaning ‘The tree which kills the wife’, i.e. when an unfaithful wife goes to collect firewood, the tree shatters at her tough and a branch pieces her abdomen. (Bouquet and Debray, 1974). The Gbaya of the central African Republic recognize the tree as producing good firewood; indeed classified it as ‘woman’s firewood’ being good for the hearth and cooking place, long- lasting while the house wife is about other chores, yet reviving quickly from sleeping embers, with a hot flame and little smoke (Bouquet, 1969). The tree is used for house- posts in southern Nigeria, and in Gabon where the wood is made into charcoal for blacksmith’s work. In Kenya and in Uganda, the wood is known for its hardness, denseness, durability and good resistance to termite- attack. It is used to make pestles and back- cloth mallets. Charcoal made from the branches is powdered and rubbed on the head for headache in the soudanian region (Burkill, 1985). The foliage is browsed a little by cattle in Senegal as a supplementary food and in Nigeria. Leaves and leafy shoots have a considerable medicinal use. When chewed, they have an acid taste. Leaves are prepared into infusion for use in Senegal for chest complaints and small- pox and with the roots, for deficiency diseases. A macerate is given for gripe, and leaf- decoction used as an eye wash (Daziel, 1937). A decoction with honey is taken in Guinea for biliousness. In Ivory coast- upper Volta, a leaf- decoction is used in baths and draughts as a febrifuge and leaf- powder is taken as snuff for headache or applied topically for rheumatic pains and toothache, or for the same purposes, leaves may be pulped with an organic acidic substances such as citron juice or sap of Piliostigma reticulatum (leguminosae: caesalpinioideae), (Kerharo, 1974).

35

P.N. Olotu et al: Continental J. Pharmaceutical Sciences 4: 35 - 39, 2010

Justification The Hausa tribe in the Northern Nigeria has over the years used the decoction of leaves and the stem bark or root bark of Hymenocardia acida in the treatment of pain of various categories such as migraine, sickle cell crisis and menstrual pain (Agishi, 2004). So far, no official work has been done on the toxicity and haematological studies of the root-bark extracts to establish its safety scientifically. This study aims at establishing the safety of the crude drug with the view of its future development. MATERIALS AND METHODS Plant Collection and Identification: The root bark of Hymenocardia acida was collected from Kudingi village, Zaria, Nigeria. The plant was identified in the field using keys and description given in the official books (Woody Plant of West Tropical Africa). The collection (voucher specimen) was confirmed and authenticated at the Herbarium, Biological Sciences, Ahmadu Bello University (A.B.U.) Zaria (1010). Acute Toxicity Studies The Lorke method (1983) was used for this study. Doses of 500, 1000, 2000, 3000, 4000 and 5000 Mg/Kg body weight (bw) and 8 mice were used. Doses of 10,000 and 11,000 Mg/Kg bw and 3 mice were used orally. Chronic Toxicity Studies The use of Hymenocardia acida in traditional medicine is normally a long term administration and this is why the aim of this study is to monitor the long term effect of this drug on the animal tissue and organs. Male Swiss albino mice aged 4-6 weeks were used and these were grouped into 3 of 6 mice each. The control group (1) was given un-amended diet and water. Whereas the treated (2 & 3) groups were given amended diets of 25%w/w and 50%w/w of the plant root bark respectively for 3 months. The pre and post-treatment water-intake and body weights of each mouse were daily recorded. At the end of the 3 months, the mice were sacrificed and their various organ weights recorded. The organs checked for histopathology include the liver, the kidney, the heart, the lung and the skin. The organs were fixed in 10% buffered neutral formalin for 72h before processing. This was an attempt to maintain the tissues as it was in the ante- mortem before the post- mortem. The different tissues were then labelled and allowed to dehydrate in graded series of alcohol in the ascending order of 70%, 80%, 95% and 100% alcohol after which the tissues were cleared with xylene and impregnated with paraffin wax, separately embedded for sectioning with rotatory microtome and microtome knife. The tissues were then sectioned at 6- micro thick and were mounted on a clean and grease free slip and then dried in an oven. The stained slides were examined with the compound microscope at X40 objectives and the results were recorded. This was repeated with the other 6 mice fed with 50% w/w amended diet and the control group (guided by the Department of Pharmacology, Ahmadu Bello University, Zaria). This experiment was repeated for female albino mice. Haematology The experiment was aimed at evaluating the effects of the plant on blood parameters with the view of investigating the traditional claims of the plant by the Hausas in the treatment of sickle cell anaemia. The parameters observed were full blood count or differential count for complete blood count (CBC), packed cell volume (PCV), mean haemoglobin concentration (MHC), total red blood cell count (RBC count), mean corpuscular volume (MCV), total white blood cell count (WBC count), differential Leucocyte count (LC), and platelet count. Swiss albino mice were fed with amended diet containing 25%w/w and 50% w/w of the root bark. After 65 days, the mice were euthanized and the blood parameters were evaluated using standard methods described by (Akinloye and Olorede, 2004). The results were compared with animals fed on unamended diet. RESULTS AND DISCUSSION Acute toxicity studies The water extract of Hymenocardia acida when administered orally was found to be safe. No death was recorded nor was any sign of toxicity observed within 24h of administration even at a higher dose of 11,000mg/kg although LD50 of values greater than 5,000mg/kg have no therapeutic value (Lorke, 1983).

36

P.N. Olotu et al: Continental J. Pharmaceutical Sciences 4: 35 - 39, 2010

Chronic Toxicity Studies Over 90 days of the experimental studies, the mice fed with 25% w/w amended diet did not show any observable tissue damage in the lungs, hearts, kidneys, brain intestines and spleens. The control rats also showed no visible liver damage but the rats fed with 50% w/w amended diet reacted differently. They showed a focal area of hepatic necrosis and the globule cells of the intestine were covered with progressive mucin and lymphocyte proliferations were observed with the spleen. But obviously there was no tissue damage of the pancreas, hearts, lungs, kidneys and brain. This is to say that prolonged (chronic) treatment with this drug in traditional medicine should be cautioned. Furthermore, the body weight, feed and water-intake increase with the mice fed with the lower concentration of the amended diet (25% w/w) but decreased with the higher concentration (50% w/w). There was also a prominent change with the post treated mice. This result may suggest the use of the plant for nutritional purposes at lower concentration. The different plates (1, 2, 3 and 4) are shown below. Haematology studies Analysis of the blood parameters showed very significant increase with the RBC (Table 1) this justifies the claim of the plant in the treatment of anaemia. Analysis of the plant on the remaining blood parameters was not significant (Table 2, 3, 4, 5, 6 & 7).

Plate 1: Fatty degeneration of the Hepatic cells

Plate 2: Necrosis of the Hepatic cells

Plate 3: Progressive mucin seen on

Plate 3: Progressive mucin seen on the globule cells of the Intestine
37

Plate 4: The section of the spleen showing lymphocyte proliferation

P.N. Olotu et al: Continental J. Pharmaceutical Sciences 4: 35 - 39, 2010 Table 1: Red blood cells (RBC) Root bark ANOVA Source of Variation SS DF Between Groups 3.754167 2 Within Groups 5.883333 13 Total 9.6375 15

MS 1.877083 0.452564

F 4.147663

P-value 0.040437

F crit 3.805567

Table 2: Packed cell volume (PCV) Root bark ANOVA Source of Variation SS DF MS Between Groups 1.914167 2 0.9570833 Within Groups 7.443333 13 0.5725641 Total 9.3575 15 Table 3: Hb (Root bark) ANOVA Source of Variation SS Between Groups 331772.4 Within Groups 2669449 Total 3001222 Table 4: TP (Root bark) ANOVA Source of Variation SS Between Groups 1.914167 Within Groups 7.443333 Total 9.3575

F 1.671574

P-value 0.225917

F crit 3.805567

DF 2 13 15

MS 165886.2 205342.3

F 0.807852

P-value 0.466986

F crit 3.805567

DF 2 13 15

MS 0.957083 0.572564

F 1.671574

P-value 0.225917

F crit 3.805567

Table 5: White blood cells (WBC) Root bark ANOVA Source of Variation SS DF MS Between Groups 18.00417 2 9.002083 Within Groups 140.8133 13 10.83179 Total 158.8175 15 Table 6: Neutrophils (Neu) Root bark ANOVA Source of Variation SS DF Between Groups 21.02083 2 Within Groups 155.4167 13 Total 176.4375 15 Table 7: Lymphocytes (Lymp) Root bark ANOVA Source of Variation SS DF Between Groups 30.52083 2 Within Groups 172.4167 13 Total 202.9375 15

F 0.83108

P-value 0.457452

F crit 3.805567

MS 10.51042 11.95513

F 0.879155

P-value 0.438423

F crit 3.805567

MS 15.26042 13.26282

F 1.150616

P-value 0.346665

F crit 3.805567

CONCLUSION The toxicity and haematology studies of the root bark of Hymenocardia acida have been carried out successfully. The results obtained could serve as the tool for establishing the safety of the use of this plant in traditional medicine and in anaemia.

38

P.N. Olotu et al: Continental J. Pharmaceutical Sciences 4: 35 - 39, 2010

REFERENCES Agishi, E. C., (2004). Etulo, Idoma, Igede, Tiv and Hausa names of Plants. AGITAB Publishers Ltd. Makurdi, Nigeria. pp.188. Akinloye, J., & Olorede, M. (2004). Toxicity study of the Leaves of Cochlospermum planchonii. Vol. 13. Pp 23. Bouquet, A., & Debray, M. (1974): Medicinal plants of the Ivory Coast, Trav. Doc. Orst 32: (Serv. Cent. Document Orstambondy 93140 France) pp.1 Bouquet, A. (1969). Medicinal plants of the Ivory Coast, Trav. Doc. Orst pp 116.
nd

Burkill, H. M., (1985). The Useful Plants of West Tropical Africa, 2 Edition, Vol.1. Royal Botanic Gardens, Kew, 960p. Daziel, J. M., (1937). Useful plants of Tropical West Africa. Crown Agent for Overseas Government and Administration, London. pp 52-53. Kerharo, J. (1974). Historic and Ethnopharmacognosic review on the belief and Traditional practices in the treatment of sleeping sickness in West Africa, Bull Soc. Med. Afri. Noire Lang. FR. 19: (Fac. Med. & Pharm Dakar Senegal) pp. 400. Lorke, D.A. (1983). A New Approach to Practical Acute Toxicity Testing. Arch. Toxicology 54:275-287. Received for Publication: 07/09/2010 Accepted for Publication: 19/10/2010 Corresponding Author: J.S Gushit Department of Science Laboratory Technology, Faculty of Natural Sciences University of Jos, Nigeria. Email: johngushit@yahoo.com

39