DDVP(O,O-dimethyl-O-2,2-dichlorovinylphosphate)isanorgano-phosphateinsecticideusedincropandfoodstorageareas,greenhouses,barnsandinworkplaces.ThepresentstudyexaminedhematologyofAlstoniabooneistembarkextractinDDVP-inducedexperimentalrats.Twentymalerats’weightranges100-190gdividedintofourgroupsoffiveratseach.GroupAwerecontrolrats,groupBreceivedDDVPonly,groupCreceivedDDVPand200mg/kgdosageofextractandgroupDreceivedDDVPand400mg/kgdosageofextract.Increasingdoses(200and400mg/kgbodyweight)ofAlstoniabooneiethanolstembarkextractwereadministeredbyoralgavagetotheothertwotreatmentgroupCandDfor21days.Theanimalsweresacrificedusingdiethylether,andtheirbloodsamplecollectedintoEDTAbottles,forassessinghemoglobin(Hb),redbloodcell(RBC),packedcellvolume(PCV),whitebloodcell(WBC),lymphocyte(L),monocytes(M),eosinophils(E),neutrophils(N),Meancorpuscularvolume(MCV),meancorpuscularhemoglobin(MCH)andmeancorpuscularhemoglobinconcentration(MCHC).StatisticalanalysesoftheresultsshowssignificantdecreaseinthehemoglobinandreverseallabnormalitiesinthehematologicalparameterswhencomparedDDVPcontrolwiththeextractstreatedgroups.Inconclusion,AlstoniabooneiethanolicstembarkhasaprotectiveeffectonbloodprofileagainsttheDDVPtoxicity

Ojo, A. Oluwafem

Ojo, B. Adebola

Afe Babalola University Repository

Archives • 2014 • vol.3 •136-140EFFECTS OF ALSTONIA BOONEI EXSTRACT ON HEMATOLOGICAL INDICES OF MALE WISTER RATOjo, A. Oluwafemi1* ; Ojo, B. Adebola21Department of Chemical Sciences, Biochemistry unit, Afe Babalola University, Ado Ekiti, Ekiti State, Nigeria.2Department of Biochemistry, Ekiti State University, Ado-Ekiti, Ekiti State, Nigeria.* oluwafemiadeleke08@gmail.com.AbstractDDVP (O, O-dimethyl-O- 2, 2-dichlorovinyl phosphate) is an organo-phosphate insecticide used in crop andfood storage areas, greenhouses, barns and in workplaces. The present study examined hematology ofAlstonia boonei stem bark extract in DDVP-induced experimental rats. Twenty male rats’ weight ranges 100-190 g divided into four groups of five rats each. Group A were control rats, group B received DDVP only,group C received DDVP and 200 mg/kg dosage of extract and group D received DDVP and 400 mg/kg dosageof extract. Increasing doses (200 and 400 mg/kg body weight) of Alstonia boonei ethanol stem bark extractwere administered by oral gavage to the other two treatment group C and D for 21 days. The animals weresacrificed using diethyl ether, and their blood sample collected into EDTA bottles, for assessing hemoglobin(Hb), red blood cell (RBC), packed cell volume (PCV), white blood cell (WBC), lymphocyte (L), monocytes (M),eosinophils (E), neutrophils (N), Mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) andmean corpuscular hemoglobin concentration (MCHC). Statistical analyses of the results shows significantdecrease in the hemoglobin and reverse all abnormalities in the hematological parameters when comparedDDVP control with the extracts treated groups. In conclusion, Alstonia boonei ethanolic stem bark has aprotective effect on blood profile against the DDVP toxicity..Keywords: Hematology, Alstonia boonei, DDVPDecember 30, 2014_______________________________________http://pharmacologyonline.silae.itISSN: 1827-8620IntroductionAlstonia boonei De Wild is a largedeciduous evergreen tree, of about 45 mhigh and 1.2 m in diameter, belonging tothe family Apocynaceae which consists ofabout 40-60 species. It is a native of tropicaland subtropical Africa, Southeast Asia, CentralAmerica and Australia. ‘Alstonia’ is namedafter Dr C. Alston (1685-1760). It’s called‘devil tree’ in tropical and subtropical Africa;Central America and Australia. It is reportedlyused as treatment of malaria, intestinalhelminthes, rheumatism, muscular pain,insomnia, and hypertension. It is known tocontain phytochemicals such as saponins,alkaloids, tannins and steroids [1, 2]. In folkmedical practice, an infusion of the stembark extract serves as antisnake venom andas antidote to some arrows poisons.Antimalarial potentials of various fractions ofthe stem bark extract of Alstonia boonei wasreported by [3, 4] while [5, 6, 7] confirmedindigenous medicinal usefulness of Alstoniaboonei for malaria therapeutic use inSouthwestern part of Nigeria. Theerythrocytes (red blood cell) are a nucleatepacked with the oxygen carrying protein-hemoglobin; the cell survive in the circulationfor about 120 days, worn out erythrocytesremoved from circulation by the macrophagesof the spleen and bone marrow. The signal forremoval is by complex oligosaccharidesattached to integral membrane protein of theplasmalemma [8]. Dichlorvos or 2,2-dichlorovinyl dimethyl phosphate (TradeNames: DDVP, Vapona, etc.) Is a volatileorganophosphate, used as an organo-phosphorus insecticide to control householdpests, in public health, and protecting storedproduct from insects.]. Dichlorvos has high toextreme acute toxicity from oral or dermalexposure, and extreme acute toxicity frominhalation. Taking in large doses may causenausea and vomiting, restlessness, sweating,and muscle tremors. Large doses may causecoma, inability to breathe, and death [9].Because of the widespread use of Alstoniaboonei, it is necessary to study its effect onblood, the [8]. To further carry out scientificscrutiny on this plant, this study examined thehematological responses of the stem barkextract of A. boonei in rats since blood areassociated with the development of several diseases.Materials and MethodsPlant material Fresh stem bark of Alstonia boonei was collected onthe 16th March, 2014 from a local garden at Ekiti State,Nigeria. The plant was identified and authenticated atthe Department of Plant Science, Ekiti StateUniversity, Ado-Ekiti, Nigeria.Preparation of Alstonia boonei stems bark extractFresh stem bark of A. boonei was cut into small piecesand air-dried at room temperature. Five hundredgram (500 g) of the air-dried stem bark of the plantwas milled into fine powder using commercialblender. The powdered stem bark was extracted withone liter (1L) of 70% ethanol for 12hours bymaceration method. The ethanolic extract wasconcentrated to dry under reduced pressure at50±1oC in a rotator evaporator. The resulting crudedark-brown powdered residual extract gave apercentage yield of 5.01%.AnimalsTwenty (20) inbred albino male rats (Rattusnorvegicus) weighing between 100-190 g was used inthe study. The animals were obtained from theanimal house of the Department of ChemicalSciences, Afe Babalola University, Ado-Ekiti, Nigeria.The animals divided into cages and allowed toacclimatize for 7 days in a well-ventilated room ata room temperature of 25.0±2.0°C under naturallighting condition. The animals were allowed freeaccess to standard rat chow (Topfeeds Ltd., Ado-Ekiti,Nigeria) and distilled water ad libitum. All animals inthis study follow the international, national andinstitutional guidelines for Care and Use of LaboratoryAnimals as published by the [10].Induction of DDVPDDVP [O,O-dimethyl O-(2,2-dichlorovinylphosphate)induced in groups B and C. Briefly, DDVP dissolved indistilled water and after that managed by intravenousinjection (through tail vein) at a dose of 50 mg/kgbody weight.Experimental DesignTwenty rats with fifteen DDVP induced rats and fournormal rats, divided into four groups with five ratseach._______________________________________http://pharmacologyonline.silae.itISSN: 1827-8620PhOL Ojo & Ojo 137 (136-140) Group A: Control ratsGroup B: DDVP ratsGroup C: DDVP rats 200 mg/kg body weight ofAlstonia boonei stem bark extractGroup D: DDVP rats 400 mg/kg body weight ofAlstonia boonei stem bark extract according tomethod of [11].Blood sample collectionWhole blood was collected by cardiac puncturefrom each experimental rat into a tube containingEthylenediaminetetraacid (EDTA).Hematological parameters determinationThe parameters such as packed cell volume (PCV),hemoglobin (Hb), white blood cell (WBC), red bloodcell (RBC), neutrophil, lymphocytes, eosinophils,monocytes, mean corpuscular hemoglobin (MCH),mean corpuscular hemoglobin concentration(MCHC) and mean corpuscular volume (MCV)analyzed using an automated analyzer (Sysmex K-2IN, Japan)Statistical analysisThe data were analyzed statically using students’ T-test and one-way ANOVA. Values of p < 0.05considered significant.ResultTable 1 shows significant decrease in thehaemoglobin (Hb), packed cell volume (PCV), meancorpuscular haemoglobin concentration (MCHC),mean corpuscular haemoglobin (MCH), meancorpuscular volume (MCV), red blood cell (RBC) inDDVP-induced untreated group when compared tocontrol, 200 mg/kg and 400 mg/kg treated groups.group fed with normal diet as depicted in figure 2.DiscussionThe present decade witnessed a great andintense resurgence in the interest and use ofmedicinal plant and medicinal plant products,especially in Africa and North America. The usefuleffects of these plant materials ascribed to thecombinations of secondary metabolites present inthe plant [12]. Botanical medicine has beenrecognized has one of the oldest practicedprofessions by humanity [13]. Apart from the lack ofinformation about the adverse or toxicity effect ofthis plant; its widespread use in folk medicinal ortraditional practice provides lack of information.underlying the biochemical mechanism responsiblefor some of the observable and reported propertiesof this plant. The result of the present studysuggests that A. boonei stem bark extract caused asignificant decrease in hemoglobin of the rats.However, hematological parameters provideinformation on the status of bone marrow andhemolysis [14]. Also, it’s revealed by the presentstudy that hematological parameters in DDVP control(untreated group) showed abnormalities. This mightresults in glycosylated hemoglobin with decrease inred blood cell (RBC). This suggests an imbalancebetween its synthesis and destruction and packedcell volume (PCV) normally affected by DDVP-induced toxicity, a sign of anemia [15, 16]. LoweredRBC count, decreased MCH and MCV are otherhematological changes found in the group whichDDVP administrated [15, 17]. Anemia was in theform of microcytic and hypochromic. This might bebecause of effects of dichlorvos in cell metabolism,changes of the enzyme, with reactions in whichcalcium is their secondary mediator. In addition,decrease in the MCV, MCHC and MCH relates al redblood cells while decrease in WBC and its indices(lymphocytes, neutrophil, monocytes) DDVP control(untreated group) might suggests decrease inimmune in fighting foreign substances. Theperformance of the ethanolic extract (especially at400 mg/kg) in reversing this irregularities in thehematological parameters may be due ascribes tothe presence of iron in the plant extract as reportedby [18], an essential part of many enzymes in cellsand parts of heme group in hemoglobin. Most iron inthe body stored within the red blood cells where ironis critical for hemoglobin synthesis [14]. Thepresence of other antioxidant vitamins (vitamin B,and C) and mineral (Zn, Ca, Fe, K, P, Cu etc), totalflavonoids and total phenol [18] might be responsiblein improving the immune being weak because of thegeneration of reactive oxygen species because ofDDVP-induction and shows the ethanolic extract maynot have negative effect on the bone marrow andhemoglobin metabolism.ConclusionIn conclusion, Alstonia boonei ethanolic stem bark has a protective effect on blood profile against the DDVP toxicity.Conflict of InterestThe authors declare no conflict of interest. Noexternal funding._______________________________________http://pharmacologyonline.silae.itISSN: 1827-8620PhOL Ojo & Ojo 138 (136-140) References1. Taiwo OB, Kroes BH, Beukelman CJ, Horsten S, HorstenS, Horsten S, Makinde, JM, Labadie RP. Activity of thestem bark extract of Alstonia boonei de Wild(Apocynaceae) on human complement and Apolymorphonuclear leukocytes. Ind. J. Pharmacol. 1998,30: 169-174.2. Osadebe PO. Anti-inflammatory properties of the rootbark of Alstonia boonei. Nig. J. Nat. Prdt. Med. 2002, 6:39-41.3. Bello IS, Oduola T, Adeosun OG, Omisore NOA, RaheemGO, Ademosun AA. Evaluation of antimalarial activity ofvarious fractions of Morinda lucida and Alstonia booneistem bark. Global J. Pharmacol. 2009, 3(3): 163-165.4. Iyiola OA, Tijani AY, Lateef KM. Antimalarial activity ofethanolic stem bark extract of Alstonia boonei in mice.Asian J. Biol. Sci. 2011, 4: 235-243.5. Odugbemi TO, Akinsulire, OR. Medicinal plants useful formalaria therapy in Okeigbo, Ondo State, Southwest,Nigeria. Afr. J. Trad. CAM. 2007, 4(2): 191-198.6. Idowu OA, Soniran OT, Ajana O, Aworinde DO.Ethnobotanical survey of antimalarial plants used in OgunState, Southwest, Nigeria. Afr. J. Phar. Pharmacol. 2010, 4:055-060.7. Gbadamosi IT, Moody JO, Lawal AM. Phytochemicalscreening and proximate analysis of eightethnobotanicals used as antimalaria remedies in Ibadan.Nig. J. Appl. Biosci. 2011, 44: 2967-2971.8. Janqueira LC, Carneiro J. Basic Histology text and atlas,10th edition. Mc Grawhill, New York, pp 223-237. 2005.9. Unni LK, Hannant ME, Becker RE. High-performance liquidchromatographic method using ultraviolet detection formeasuring metrifonate and dichlorvos levels in humanplasma. J chromatogr. 1992, 573, 99-10310. Canadian Council on Animal Care. The Care and Use ofFarm Animals in Research, Teaching and Testing CCAC.,Ottawa, ON. 2009;12-15.11. Ojo OA, Ajiboye BO, Oyinloye BE, Ojo AB. Prophylacticeffects of Ethanolic extract of Irvingia gabonensis stem barkagainst Cadmium-induced Toxicity in albino rats. Advancesin Pharmaceutics, 2014; Article ID 894610, 8 pages, HindiwiPublishing Co-operation.12. Briskin DP. Medicinal Plants and Phytomedicine. LinkingPlant Biochemistry and Physiology to Human Health.Plant Physiology, 2000, 124: 507-514.13. Oduola T, Adeniyi FAA, Ogunyemi EO, Bello IS, Idowu TO,Subair HG. Toxicity studies on an unripe Carica papayaaqueous extract: Biochemical and haematological effects inwister albino rats. 2007.14. Iniaghe OM, Egharevba O, Oyewo EB. Effects of aqueousleaf extract of Acalpha wilkesiana on hematologicalparameters in male wistar albino rats. British journal ofpharmaceutical research. 2013, 3(3): 456-471.15. Falke HE, Zwennis WCM. Toxicity of lead acetate to femalerabbits after chronic subcutaneous administration:Biochemical and clinical effects. Arch. Toxicol.64: 1990, 522-529.16. Aladodo RA, Muhammad NO, Balogun EA. Effects ofaqueous root extract of Jatropha curcas on hyperglyceamicand haematological indices in alloxan-induced diabetic rats.Fountain Journal of natural and applied sciences, 2013,2(1). 52-58.17. Yagminas AP, Franklin CA, Villeneuve DC, Gilman AP, LittlePB, Valli VE. Subchronic oral toxicity of tri-ethyl lead in themale weanling rat. Clinical, biochemical, hematological, andhistopathological effects. Fundam.Appl. Toxicol. 1990, 15:580-596.18. Akinmoladun AC, Ibukun EO, Afor E, Akinrinola BL, OnibonTR, Akinboboye AO, Obuotor EO, Farombi EO. Chemicalconstituents and antioxidant activity of Alstonia boonei.Afr. J. Biotechnol. 2007, 6(10): 1197-1201._______________________________________http://pharmacologyonline.silae.itISSN: 1827-8620PhOL Ojo & Ojo 139 (136-140) _______________________________________http://pharmacologyonline.silae.itISSN: 1827-8620PhOL Ojo & Ojo 140 (136-140) Groups Hb (g/dl) PCV (%) MCHC (X102g/l) MCH (pg) MCV (fl) RBC (X1012/L)Control 15.6 ± 0.68a 37.8 ± 1.07 ab 3.12 ± 0.05a 29.60 ± 0.68a 78.60 ± 2.16a 5.34 ± 0.24bDDVP untreated 11.1 ± 0.05b30.23 ± 0.40c 1.21 ± 0.02b 20.14 ± 0.23b 48.45 ± 0.02b 2.50 ± 0.02c200mg/kg treated 14.8 ± 0.58a 39.8 ± 0.58b 3.18 ± 0.03a 28.60 ± 0.51a 74.80 ± 2.18a 4.74 ± 0.12a400mg/kg treated 15.00 ± 0.71a 35.00 ± 1.00a 3.12 ± 0.03a 29.20 ± 0.73a 78.20 ± 1.56a 4.26 ± 0.05aTable 1: Effects of A. boonei stem extract on hematological parameters in Dichlorvos induced Wister rats.g/dl = gram per deciliter, MCH = mean corpuscular hemoglobin, pg = Picogram, MCV = mean corpuscular volume, fm=femtoliter. Values are expressed as mean of five replicates ± SEM. Values with different superscripts (abc) along therow are significantly different (p<0.05)GroupsWBC(X103mm2)Neu (%) Lym (%) Eos (%) Mon (%)Control 4.16 ± 0.12b 49.40 ± 0.75ab 41.80 ± 0.80b 2.80 ± 0.37a 6.00 ± 0.63aDDVP untreated 1.2 ± 0.01c 31.02 ± 0.03c 32.10 ± 0.01c 0.2 ± 0.31b 3.2 ± 0.31b200mg/kg treated 4.30 ± 0.25b 47.80 ± 0.20a 42.60 ± 1.12b 2.2 ± 0.20a 7.4 ± 1.12a400mg/kg treated 2.96 ± 0.12a 50.40 ± 0.51b 39.00 ± 0.45a 3.4 ± 0.51a 7.2 ± 0.66aTable 2: Effects of A. boonei stem extract on White blood cell indices in Dichlorvos induced Wister rats.WBC = White blood cell, Neu = Neutrophils, Lym = Lymphocytes, Eos = Eosinophils, Mon =Monocytes. Values are expressed as mean of five replicates ± SEM. Values with differentsuperscripts (abc) along the row are significantly different (p<0.05)

EFFECTS OF ALSTONIA BOONEIEXSTRACT ON HEMATOLOGICAL INDICES OF MALE WISTER RAT

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EFFECTS OF ALSTONIA BOONEIEXSTRACT ON HEMATOLOGICAL INDICES OF MALE WISTER RAT

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