Aqueous Extract Of Vernonia Amygdalina Protects Against .
OPENACCESSWorld J Young Researchers 2012;2(5)2(5):ISSN 2249-05662(5):70Ogunlade et al., 2012. Vernonia amygdalina Protects Against Hepatotoxicity in Wistar RatsAqueous Extract of Vernonia amygdalinaProtects Against Alcohol-InducedHepatotoxicity in Wistar RatsOriginalArticleLife ScienceBabatunde OGUNLADE 1, Godson G AKUNNA 1, Oluwaseun O FATOBA 2,Omolara J AYENI 2, Adebiyi A ADEGOKE 2, Sunday A ADELAKUN 2ABSTRACT ons:Adverse health effects have been recognized to be caused by Alcohol has been shown to cause adverse health effectworldwide; and liver is one of the organs mostly affected. The effect ofVernoniaamygdalina against alcohol-inducedhepatic damage in rats was investigated in the present study. Rats were divided into three groups; the alcohol-alonegroup was given 5 ml/kg body weight of 56% ethanol (v/v) daily per oral for 35 days.The alcohol plus V. amygdalinagroup were similarly given alcohol, but had V. amygdalina 300 mg kg-1 b.wt, daily p.o post-treatment for another 35 days.Another group of rats were given distilled water (the vehicle) 300 mg kg-1 b.wt, daily p.o, for 35 days to serve as thecontrol. The gross anatomical parameters of the liver and liver histology were assessed. Liver oxidative stress wasevaluated by liver Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione(GSH) and malondialdehyde (MDA) assays. In addition, the activities of the biomarker enzymes of the liver (alaninetransaminase, aspartate transaminase and alkaline phosphatase were assayed. An assessment of the histological profilesof the liver showed a derangement of the liver cytoarchitecture following alcohol abuse and a marked improvement wasobserved after V. amygdalina administration. Similarly,V. amygdalinaimproved the reduction of antioxidant parameters(SOD, CAT, GPx and GSH) and the increased MDA caused by alcohol ingestion. It was concluded that V. amygdalinamayoffer protection against free radical mediated oxidative stress of rats with alcohol-induced hepatotoxicity.Keywords: Alcohol, liver, oxidative stress, antioxidants, Vernonia amygdalinaRÉSUMÉ [FRANÇAIS/FRENCH]FRANÇAIS/FRENCH]Effets néfastes sur la santé ont été reconnus pour être causés par l'alcool a été montré pour causer des effets néfastessur la santé dans le monde entier et le foie est un des organes les plus touchés. Le ofVernoniaamygdalina effet contre lesdommages hépatiques induite par l'alcool chez le rat a été étudiée dans la présente étude. Les rats ont été divisés entrois groupes: le groupe de l'alcool seul a donné 5 poids corporel ml / kg d'éthanol 56% (v / v) par jour et par voie oralependant 35 days.the alcool plus V. amygdalina groupe ont même donné de l'alcool, mais il a V. amygdalina 300 mg kg-1b.wt, tous les jours po post-traitement pendant encore 35 jours. Un autre groupe de rats ont reçu de l'eau distillée (levéhicule) 300 mg kg-1 b.wt, tous les jours po, pendant 35 jours pour servir de contrôle. Les paramètres anatomiquesbrut de l'histologie hépatique et le foie ont été évalués. Le stress oxydatif hépatique a été évaluée par la superoxydedismutase foie (SOD), la catalase (CAT), glutathion peroxydase (GPx), le glutathion réduit (GSH) et de malondialdéhyde(MDA) des essais. En outre, les activités des enzymes marqueurs biologiques du foie (alanine aminotransférase,aspartate transaminase et de phosphatase alcaline ont été analysés. Une évaluation des profils histologiques du foie amontré un dérangement de la cytoarchitecture du foie suite à l'abus d'alcool et une nette amélioration a été observéeaprès V. amygdalina administration. de même, V. amygdalinaimproved la réduction des paramètres antioxydants (SOD,CAT, GPx et GSH) et l'augmentation de MDA causée par l'ingestion d'alcool. Il a été conclu que la protection contre V.offre amygdalinamay stress oxydatif des radicaux libres induite chez le rat avec de l'alcool hépatotoxicité.Mots-clés: L'alcool, le foie, le stress oxydatif, antioxydants, Vernonia amygdalina1Department ofAnatomy, College ofMedicine, Lagos StateUniversity, IkejaLagos, NIGERIA2Department ofAnatomy, Faculty ofBasic MedicalSciences, College ofMedicine of theUniversity of Lagos,Idi-Araba Lagos,NIGERIA.Email Address forCorrespondence/Adresse de courrielpour ccepté:September, 2012Full Citation:Ogunlade B, AkunnaGG, Fatoba OO,Ayeni OJ, AdegokeAA, Adelakun SA.Aqueous Extract ofVernonia amygdalinaProtects AgainstAlcohol-InducedHepatotoxicity inWistar Rats. WorldJournal of YoungResearchers2012;2(5):70-7.INTRODUCTIONdisease process which typically progresses through theAlcohol liver disease (ALD) remains a major treat,stages of alcoholic steatosis, alcoholic hepatitis andcomplication and causes of increase in mortality andalcoholic cirrhosis to end-stage liver disease [2-3].morbidity rate (reaching about 6.1%) in heavy alcoholChronic alcohol consumption has been reported todrinkers [1]. ALD poses a major health and economicinduce oxidative stress via cytochrome P450 2Eiconcern and its treatment forms has remained a great[4].Numerous studies to elucidate the pathogenesis ofchallenge for researchers. ALD is a complex chronicALD have been done [5-9] and this is majorly becauseOPENACCESS Research Reviews Publications, 2011http://www.rrpjournals.com/
ISSN 2249-0566World J Young Researchers 2012;2(5)2(5):2(5):71Ogunlade et al., 2012. Vernonia amygdalina Protects Against Hepatotoxicity in Wistar RatsOPENACCESSapproximately 80% of ingested alcohol is metabolized inthe Lagos state university college of Medicine, Ikeja. Thethe liver by a process that produces numerous ROS (S).animals were housed in well ventilated wire woodenIn 2004, Lieber [10] reported that alcohol metabolismcages in the animal facility of the department ofcould also lead to production of aldehydes which containAnatomy, Lagos state University College of Medicine,a potent pro-inflammatory and profibrotic properties.Ikeja. The rats were maintained under standard naturalDamage tocell membranes and organelles has beenphotoperiodic condition of twelve hours of lightlinked to lipid peroxidation through oxidative stressalternating with twelve hours of darkness(i.e.L:D;12h:12hleading to the release of reactive aldehydes [11].photoperiod) at room temperature (25-26oC) and relativeSince indigenous plants have been the traditional sourcehumidity of 65 5%. They were allowed unrestrictedof raw materials for the manufacture of medicines,accessalternative practitioners thus routinely recommendtoacclimatizedover a period of 20 days before thenatural antioxidant supplement for ALD.commencement of the experiment. The weights of theConsiderable interest have developed in plants likeanimalsVernonia amygdalina, Del (Compositae), which is a well-acclimatization, at commencement of the experimentsknown herbal medicine widely used as food with largeand whenthe experiment was completed using anvariety of beneficial effects. V. amygdalina is a well-electronic analytical and precision balance (BA210S,recognised African medicinal plant whose tree is about 1-d 0.0001g) (Satorius GA, Goettingen, Germany).3 m in height [12-14]. It is a medium sized shrub whoseExperimental procedures involving the animals and theirgreen leaf is about 6mm diameter and commonly calledcare were conducted in conformity with international“bitter leaf” and “ewuro” among Yoruba people innational and institutional guidelines for the care ofsouthwest Nigeria. It has been reported to containlaboratory animals in biomedical research and use ofantioxidants (luteolin, luteolin 7-O- -glucuronoside andlaboratory animals in bio-medical research promulgatedluteolin 7-O- -glucoside flavonoid) isolated from theby the Canadian council of animal care curement,duringleaves [15].Several reports on the antitumor [16-22],antihelmintics [23-25], antidiabetic [26], hypolipidemicAcute Oral Toxicity Study of Vernonia amygdalina[27-29], antimalarial [30-32], antiviral [33-34] and laxatingExtract[35] activities of V. amygdalina has been documented.The acute oral toxicity study for V. amygdalina extract washThe present investigation was therefore designed toconductedexplore the hepatoprotective potential of aqueous extractCooperation and Development (OECD) [37] Guidanceof V. amygdalina leaves on alcohol induced liver damageDocument on Humane End points that should reduce thein rats.overall suffering of animals used in this type of toxicityusingtheOrganizationforEconomictest. The test used was the limit dose test of the up andMATERIALS AND METHODSdown procedure.Vernonia amygdalina and the Aqueous ExtractionBriefly, 5 animals were weighed and individuallyProcedureidentified. The first animal was given the test dose – V.The leaves of Vernoniaamygdalinaused in this researchamygdalina extract 2000 mg per kg body weight. Thewere collected from a grown plants located at a villagesecond and third animals were concurrently dosed andaround Oke-Ogun in Oyo state, Nigeria on February 1,the fourth and fifth animals sequentially dosed.2012. They were transported to the laboratory ofThe results were evaluated as follows (S Survival, X Anatomy department of Lagos StateUniversity College ofdeath). The animals were observed individually at leastMedicine,wereonce during the first 30 minutes after dosing, periodicallyauthenticated by a staff in the herbarium of theduring the first 24 hours (with special attention givenDepartment of Botany, Lagos State University, Lagosduring the first 4 hours), and daily thereafter for a totalstate, Nigeria. The leaves were thoroughly washed inperiod of 14 days. All observations were systematicallysterile water and the water was then drained fromtherecorded with individual records maintained for eachleaves.animal.AnimalsAnimal Grouping and Experimental DesignTwenty four (24) adult male wistar rats were obtainedTwenty four adult male wistar rats weighing 190-230gfrom a breeding stock maintained in the animal house ofwere used for this research work. The rats wereIkeja,Lagosstate,Nigeriaand Research Reviews Publications, 2011http://www.rrpjournals.com/OPENACCESS
World J Young Researchers 2012;2(5)2(5):ISSN 2249-05662(5):72Ogunlade et al., 2012. Vernonia amygdalina Protects Against Hepatotoxicity in Wistar RatsOPENACCESSrandomly divided into three groups (A-C) of eight ratsDetermination of Serum ALT, AST and ALP Assayeach such that the weight difference between and withinThe degree of liver damage was evaluated by ALT, ASTgroups did not exceed 20 of average weight of theand ALP in serum using a commercially available kit.sample population.Detailed procedures for the above measurements wereRats in group A which served as control were givenperformed10ml/kg/day of distilled water for 35 sRats in group B (ethanol group) were given ethanoldiluted with normal saline (56%; v/v) administered orallyDetermination of Oxidative Stress parametersthrough an orogastric cannula into the stomach via theSOD activity in liver was determined according to theesophagus at a dose of 5ml/kg body weight, three times amethod described by Marklund and Marklund xyz [39]day, for 35 consecutive days.and GSH-Px activity was determined by GSH-Px assayThe appropriate quantity of alcohol was given. Thekit. Detailed procedures for the above measurementsadministrations were done once daily.were performed according to the kits’ protocol. CAT wasRats in group C (ethanol V. amygdalina group) rats wereassayed by the method described by Ferro and Chagasfed V. amygdalina (300 mg/kg body weight/day) in[40]. The non-enzymicGSH was analyzed by the methoddrinking water administered for 35 consecutive days,of Moron, Dipierre, and Mannervik.and ethanol was given simultaneously as described forthe ethanol group.Determination of liver MDA contentsLipid peroxidation was evaluated on the base of MDAAnimal Sacrifice and Sample Extractionlevel and MDA in liver was determined using theTwelve hours after the administration of the last ethanolmethod described by Jain et al., [41].dose, the rats were at the time of sacrifice first weighedand then cervical dislocation was carried out. BloodStatistical Analysissamples were collected and centrifuged at 1,500 g/min atAll data were expressed as mean S.D. Differences4 C for 10 min to obtain serum. The abdominal cavity ofbetween groups were analyzed using one-way analysiseach rat was opened up through a midline abdominalof variance (ANOVA). A value of p 0.05 was consideredincision to expose the liver. The liver was excised andto be statistically significant.weighed; the liver was weighed with an electronicanalytical and precision balance. The liver of each animalRESULTSwasAcute oral Toxicity nation. (BA 210S, d 0.0001- Sartoriusen GA,There were no deaths of rats dosed 3000 mg/kg bodyGoettingen, Germany).weight of the plants extract both within the short andlong outcome of the limit dose test of Up and DownHistological Procedures and Analysismethod (Table 1). The LD50 was calculated to be greaterThis was done as described by Ogunlade et al [38]. Briefly,than 3000 mg/kg body weight /orally.the organs were cut on slabs about 0.5cm thick and fixedin 10% formol saline for a day after which they wereMean Body Weights (g), Liver Weights (g) and Livertransferred to 70% alcohol for dehydration. The tissuesVolumes (ml), in Control and Experimental Ratswere passed through 90% alcohol and chloroform forAs showed in table 2, ethanol induced rats lost weight asdifferent durations before they were transferred into twocompared to the control group while ethanol V.changes of molten paraffin wax for 20min each in anamygdalina rats reversed the lost weight to nearly that ofoven at 570 C. Serial sections of 5µm thick were obtainedthe control. Mean liver weight in the ethanol groupfrom a solid block of tissue and were stained withdecreased by 17% compared to that of control grouphaematoxylin and eosin stains, after which they werewhile ethanol V. amygdalina group decreased by 8%passed through a mixture of equal concentration ofrevealing the restoring potential of V. amygdalina extractxylene and alcohol. Following clearance in xylene, theto the weight lost due to ethanol administration. Thetissues were oven- dried. Photomicrographs were takenrelative liver weight (﴾liver weight/body weight﴿x100)with a JVC colour video digital camera (JVC, China)was 3% in the ethanol group compared to the controlmounted on an Olympus light microscope (Olympus UKand ethanol V. amygdalina that were 4%.Ltd, Essex,UK) to demonstrate the hepatocyte.OPENACCESS Research Reviews Publications, 2011http://www.rrpjournals.com/
ISSN 2249-0566World J Young Researchers 2012;2(5)2(5):2(5):73Ogunlade et al., 2011. Vernonia amygdalina Protects Against Hepatotoxicity in Wistar RatsOPENACCESSTable 1: This table shows results of acute toxicity test for V. amygdalinaextract (up and down procedure) in ratsTest serialAnimalDose of V. amygdalinaShort termLong SSSSSSSSS Survival; REP Right ear pierced; LEP Left ear pierced; TC Tail cut; RLT Right leg tagged; LLT Left Leg tagged, I Intact ratTable 2: This table showsmean body weight (g), liver weight (g) in control and experimental rats.ParameterGroup AGroup BGroupCInitial Body weight(g)Final body weight(g)Body weight diff.(g)Liver weight (g)Liver wt./body wt. ratio215.5 4.0235.5 4.220.09.0 0.20.04216.5 2.7205.3 2.511.2*7.5 0.2*0.03215.0 3.5213.5 3.31.5**8.3 0.2*0.04Values are expressed as mean SD for n 8; *p 0.05, **p 0.001 significantly dissimilar from controlcompared to the control group, but the serum AST andAST and ALT levelsAs showed in table 3, the serum levels of AST and ALTALT activities in the ethanol V. amygdalina group werewere significantly increased in the ethanol groupsignificantly lower than in the ethanol group.Table 3: This table shows AST and ALT levels in control and experiment ratsGROUPSAST (U/l)A (CONTROL)B (ETHANOL)C(V.amygdalina ETHANOL)25.0 2.2102.2 1.2**35.5 1.6*ALT(U/l)23.5 1.0120.6 2.1**40.2 3.0*Values are expressed as mean SD for n 8; *p 0.05, **p 0.001 significantly dissimilar from controlAntioxidant Levels (CAT, SOD, GSH, GPx ) and MDAthe ethanol group. The anti-oxidant levels (CAT, SOD,LevelsGSH and GPx ) decreased significantly in ethanol groupAs showed in table 3, the MDA levels in the ethanol(**P 0.001) compared to the control group but the CAT,group increased compared with the control group butSOD, GSH and GPx levels in ethanol V. amygdalinadecrease in ethanol V. amygdalina group compared togroup decreases (*P 0.05) compared to the control group.Table 4: This table shows effects of V. amygdalina on antioxidant levels and lipid peroxidation levels in rat liverMDACATSODGSHGPx(nmol/mg protein)Group AGroup BGroup C6.9 0.69.3 1.1**7.2 0.7*(U/mg protein) (U/mg protein) (nmol/mg protein) (nmol/mg protein)17.5 3.610.2 3.0**15.2 3.3*11.5 4.05.2 2.5**11.2 3.9*7.2 0.75.0 0.6**6.7 0.5*4.0 8.01.2 4.8**3.4 6.8*Values are expressed as mean SD for n 8; *p 0.05, **p 0.001 significantly dissimilar from control Research Reviews Publications, 2011http://www.rrpjournals.com/OPENACCESS
ISSN 2249-0566World J Young Researchers 2012;2(5)2(5):2(5):74Ogunlade et al., 2011. Vernonia amygdalina Protects Against Hepatotoxicity in Wistar RatsFigure 1:This figure shows photomicrograph of group eaminotransferase (ALT) compared to the control group(Control) rat liver stained with H&E X icantly (P 0.001) these enzyme levels. The reversalof elevated serum intracellular enzyme levels by V.amygdalina extract after ethanol administration may beattributed to the stabilizing ability of the cell membranepreventing enzymes leakages. In addition, it wasreported that the reversal of increased levels oftransaminases to nearly normal predicts the restorationof hepatocytes and regeneration of hepatic lipidperoxidation levels can be used to predict the severity ofFigure 2:ethanol induced liver damage. Antioxidants enzymesThis figure shows photomicrograph of groupsuch as SOD, CAT, GSH and GPx dependently act in theB (Alcohol treated Groups) rat liver stained with H&E Xmetabolic pathways that involve free radicals. Therefore,400SOD, CAT, GSH and GPx levels decrease in liver suggestthe toxic effects of ethanol on liver functions but theadministration of V. amygdalina can counter the efficacyof ethanol on liver cells thereby blocking the decreaseantioxidants levels. Since it was proved that thesignificance of GSH in the detoxification of chemicallyreactive metabolite in drug induced toxicity afterdecrease in GSH [43-45] then we can deduce thatincreased oxidation and decrease synthesis of GSHcauses decrease in GSH levels. Therefore increase inantioxidant enzyme activities levels (SOD, CAT, GSHand GPx) after V. amygdalina extracts administrationmight contribute to the ameliorating effects of oxidativestress.This figure shows photomicrograph of GroupMDA is a known biomarker of lipid peroxidation andC (V. amygdalinavs Alcohol group) rat liver stained withoxidative stress, the increase in MDA level signifies theH&E X 400toxicFigure 3:effectsofethanolonliver[45]butthecounteractions of V. amygdalina in reducing MDA levelsuggest the potential attributes of V. amygdalina in therestoration of damaged liver tissues after ethanoladministration. Therefore, the antioxidant potential of V.amygdalina improves the liver functions by promotingantioxidant enzyme activities, thus can be recommendedas a therapeutic agent for heavy alcohol drinkers inalcohol related liver damage.Furthermore, his
Les rats ont été divisés en trois groupes: le groupe de l'alcool seul a donné 5 poids corporel ml / kg d'éthanol 56% (v / v) par jour et par voie orale pendant 35 days.the alcool plus V. amygdalina groupe ont même donné de l'alcool, mais il a V. amygdalina 300 mg kg-1 b.wt, tous les jours po post-traitement pendant encore 35 jours.
pengaruh pemberian ekstrak etanol daun afrika (Vernonia amygdalina) terhadap kadar kolesterol total tikus putih galur wistar jantan yang diberikan pakan tinggi kolesterol serta mencari dosis optimum dari ekstrak etanol daun afrika (Vernonia amygdalina) dalam menurunkan kolesterol total pada tikus putih galur wistar jantan . 1.2 Rumusan Masalah
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The study was conducted to determine the phytochemical analysis, in vitro evaluation of antioxidant and antimicrobial activity of methanolic leaf extract of Vernonia amygdalina (bitter leaf) against Staphylococcus aureus and Pseudomonas aeruginosa. The qualitative phytochemicals in the extract of V.A include tannin, alkaloids, glycosides, anthraquinone, flavonoids, phlobatanin, saponin etc .
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6 C 2017 06204317 (e) Aqueous sodium hydroxide, aqueous potassium iodide and aqueous acidified potassium manganate(VII) are added to aqueous solutions of iron(II) sulfate and iron(III) sulfate. Iron(II) ions, Fe2 , are reducing agents in aqueous solution. Iron(III) 3 ions, Fe , are oxidising agents in aqueous solution.Complete the table.
6 C 2017 06204317 (e) Aqueous sodium hydroxide, aqueous potassium iodide and aqueous acidified potassium manganate(VII) are added to aqueous solutions of iron(II) sulfate and iron(III) sulfate. Iron(II) ions, Fe2 , are reducing agents in aqueous solution. Iron(III) 3 ions, Fe , are oxidising agents in aqueous solution.Complete the table.
Dec 16, 2020 · A Novel and Simplest Green Synthesis Method of Reduced Graphene Oxide Using Methanol Extracted Vernonia Amygdalina: . lytic graphite a decade ago by Manchester university . Figure3whichwasattributedton-π transition.en- .
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