Indomethacin-induced gastric ulceration in rats: Protective roles of Spondias mombin and Ficus exasperata

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ContentslistsavailableatScienceDirect

Toxicology

Reports

j o u r n al ho me p ag e : w w w . e l s e v i e r . c o m / l o c a t e / t o x r e p

Indomethacin-induced

gastric

ulceration

in

rats:

Protective

roles

of

Spondias

mombin

and

Ficus

exasperata

Saheed

Sabiu

a,∗

_,

_Taofeeq

_Garuba

b

_,

_Taoﬁk

_Sunmonu

c

_,

_Emmanuel

_Ajani

a

_,

Abdulhakeem

Sulyman

a

_,

_Ismaila

_Nurain

a

_,

_Abdulazeez

_Balogun

c

a_{Phytomedicine,}_Food_Factors_and_Toxicology_Research_Laboratory,_Biochemistry_Unit,_Department_of_Biosciences_and_{Biotechnology,}

KwaraStateUniversity,P.M.B.1530,Ilorin,Nigeria

b_Department_of_Plant_Biology,_University_of_Ilorin,_Ilorin,_Nigeria

c_{Phytomedicine}_and_Plant_Biochemistry_Research_Laboratory,_Biochemistry_Unit,_Department_of_Biological_Sciences,

Al-HikmahUniversity,Ilorin,Nigeria

a

r

t

i

c

l

e

i

n

f

o

Articlehistory: Received9October2014 Receivedinrevisedform 30November2014 Accepted1January2015 Availableonline8January2015 Keywords:

Antacid Antioxidative Gastroprotective NSAIDS

Protonpumpinhibitor Vagotomy

a

b

s

t

r

a

c

t

This studyinvestigated the quantitative polyphenolic constituents and

gastroprotec-tive effectsof aqueousleaf extractsof Spondiasmombin and Ficusexasperata against

indomethacin-inducedgastriculcerinrats.Ulcerationwasinducedbyasingleoral

admin-istration ofindomethacin (30mg/kgbody weight).Wistarrats were pretreated with

esomeprazole(referencedrug)atadoseof20mg/kgbodyweight,S.mombinorF. exas-perataat100and200mg/kgbodyweightoncedailyfor21dayspriortoulcerinduction.At theendoftheexperiment,gastricsecretionsandantioxidantparameterswereevaluated. Weobservedthatthesigniﬁcantlyincreased(p<0.05)ulcerindex,gastricvolume, malon-dialdehydelevelandpepsinactivitywereeffectivelyreducedfollowingtreatmentwithS. mombinandF.exasperata.Theextractsalsomarkedlyattenuatedthereducedactivityof superoxidedismutaseaswellaspHandmucincontentintheulceratedrats.These ﬁnd-ingsareindicativeofgastroprotectiveandantioxidativepotentialsoftheextractswhich isalsoevidentinthedegreeof%inhibitionagainstulceration.Theavailabledatainthis studysuggestthattheextractsofS.mombinandF.exasperataprovedtobecapableof ame-lioratingindomethacin-inducedgastriculcerationandtheprobablemechanismsarevia antioxidativeandprotonpumpinhibition.

1. Introduction

Gastric ulceration is a benign lesion onthemucosal epitheliumuponexposureofthestomachtoexcessacid andaggressivepepsinactivity[1].Itisthemostprevalent gastrointestinaldisorder everknown,accounting for an estimated15mortalityoutofevery15,000complications yearlyintheworld[2,3].Inspiteoftherapidlychanging

∗ Correspondingauthor.Tel.:+2348038179476. E-mailaddress:saeed.sabiu@gmail.com(S.Sabiu).

conceptofgastric ulcermanagementfromconventional vagotomy,prostaglandinanalogs,H2receptorantagonists andantacidstoprotonpumpinhibitors,gastrointestinal toxicity remains an impediment totheir application in clinical practice. Speciﬁcally,gastrointestinal toxicity of nonsteroidalanti-inﬂammatorydrugs(NSAIDs)originmay beas highas 4–8% per yearand thecomplications are even higher for those with additional risk factors such as prior history of ulcer disease [4]. Various synthetic antiulcerdrugsarepresentlyavailableandsomeofthese likecimetidine, misoprostol,ranitidine, omeprazoleand esomeprazoleareemployedtomanage and cureNSAID http://dx.doi.org/10.1016/j.toxrep.2015.01.002

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inducedgastriculcer.However,eachofthesedrugsconfers simplertoseveresideeffects,promptingasearchfor non-toxic,easilyaccessibleandaffordableantiulcermedication [5,6].Investigationonthephytotherapyofmedicinalplants thatarehighlyvaluedandwidelyusedinthetraditional systemsofmedicinemightprovideefﬁcientformulation forbettermanagement.SpondiasmombinandFicus exas-peratabelongstothisclassoftherapeuticplants.

S. mombin(SM) commonly knownas “Iyeye” in the South-WesternpartofNigeriaisafructiferoustreeinthe FamilyAnacardiaceae.Theplantgrowsinrainforestsand coastalareas,attainingaheightof15–22m[7].Itis com-monlyusedinfolkmedicinetocuremanydiseasesdueto itspotentbioactiveprinciplesincludingtannins,saponins, flavonoids, phenolics and anthraquinone glycosides [8]. Antioxidantvitamins;alpha-tocopherolandascorbicacid have been detectedin itsleaves extracts [9]. Tea from itsflowersandleavesistakenasananalgesicand anti-inflammatorycureagainststomachacheanddiscomfort [10].Ayokaetal.[7]havealsoreporteddecoctionfromits leavestobetherapeuticagainsturethritis,cystitisaswell aseyeand throatinflammations.ThegumfromSMhas alsobeenexploitedasanexpectorantandvermifuge.The leafextractoftheplanthasbeenoutstandinglyadvocated foruseinspeedywoundhealingprocesses,hemorrhoids andinflamedmucousmembraneduetoitstannincontent [11].Itspharmacologicalpotenciessuchasantioxidative, antimicrobial,antimalarialandantibacterialhavealsobeen documented[8,10,12,13].

F. exasperata Valh (FE), called “Epin”, “Anwerenwa” and“Kawusa”respectivelyamongtheYorubas,Igbosand Hausas in Nigeria, is commonly known as ‘sand paper tree’belongingtoMoraceaeFamily.Phytochemical anal-ysisoftheleafextractofFEhasrevealedthepresenceof ﬂavonoids,tannins,saponnins,alkaloids andcyanogenic glycosides [14]. Its medicinalefﬁcacy in treating many diseases has been researched. For instance, the South-WesternpeopleofNigeriausesthedecoctionandinfusion ofFEleavesforthemanagement,controlandtreatmentof hypertension,diabetesmellitusandcertain cardiovascu-lardysfunction[15].LeavesofFEcookedwithbananasare eatenforthetreatmentofgonorrhea[16].Itsleafextractis alsotakentosuppressstomachache,treatpepticulcerand asantidotetopoison[5].

WiththeremarkableattributesofSM andFE partic-ularlyin alleviatingstomachache relateddisorders and woundhealingenhancement,thepresentstudycompared theirtherapeuticefﬁcacytoareferencedrug (esomepra-zole)onindomethacin-inducedgastriculcerationinrats. 2. Materials

2.1. Chemicalsanddrugs

Indomethacinandesomeprazolewererespectively pro-cured from Kapit Pharmaceutical Limited, Nigeria and Ranbaxy Laboratories, India. Trichloroacetic acid (TCA), dimethylaminobenzaldehyde,epinephrine,acetylacetone, bovineserumalbumin(BSA),gallicacid,aluminum chlo-ride,quercetinandthiobarbituricacid(TBA)wereproducts ofSigmaChemicalCo.(St.Louis,MO,USA).Distilledwater

wasobtainedfromBiochemistryLaboratory,KwaraState University,Malete,Nigeria.Assaykitsusedwerefrom Ran-doxLaboratorieslimited,UnitedKingdom.Otherchemicals usedwereofanalyticalgradefromreputablecompaniesin theworld.

2.2. Plantcollectionandauthentication

FreshleavesofSMandFEwerecollectedinApril2014 followingidentiﬁcationofthetwoplantsatthebotanical gardenof Universityof Ilorin,Ilorin,Nigeria.Theleaves wereauthenticatedattheUniversity’sHerbarium,where voucher specimens (nos.14/20567and 14/20568) were preparedanddeposited.

2.3. Experimentalanimals

Albino ratsoftheWistar strainat amean weightof 180.00±1.85gwereusedforthestudy.Theanimalswere obtained and reared as described by Sabiu et al. [17], following approval from the Independent Ethical Com-mitteeontheUseandCareofLaboratoryAnimalsofthe KwaraStateUniversity,Malete,Nigeria.Acertiﬁed num-berKSU/IECCULA/001/05/014wasassignedandissuedfor theresearch.

3. Methods

3.1. Preparationofextracts

LeavesofSMandFEwereair-driedatroom tempera-turefor10daystoconstantweight.Thedriedsampleswere thenpulverizedwithanelectricblender(modelMS-223; Blender/MillerIII,Taiwan,China),weighedandkept air-tightpriortoextraction.Powderedsamples(500geach) of both plants were separately extracted in 5l of dis-tilledwater for48hwithcontinuousshaking byorbital shaker maintained at 300rpm. The solutions obtained werethenfiltered(withWhatmanNo.1filterpaper)and theresultingfiltrateslyophilizedtogive15.5g(SM)and 12.4g(FE)residues,correspondingtoyieldsof3.1%and 2.48% respectively. The lyophilizedsamples were sepa-ratelyreconstitutedindistilledwatertogivedosesof100 and 200mg/kg body weightofeach extract usedinthe study.

3.2. Determinationoftotalphenolics

FollowingthereportedmethodofWolfeetal.[18],the total phenolcontents in the plant extractswere deter-mined.Brieﬂy,analiquotofeachextract(1ml)wasmixed with5mlFolin-Ciocalteureagent(previouslydilutedwith water1:10v/v)and4ml(75g/l)ofsodiumcarbonate.The tubes were vortexed for 15s and allowed to stand for 30minat 40◦C for color development.Absorbancewas readat765nmusingaspectrophotometer(Beckman,DU 7400,USA).Extractswereevaluatedataﬁnalconcentration of1mg/ml.Totalphenoliccontentwasexpressedasmg/g gallicacidequivalentusingtheequationobtainedfroma calibrationcurveofgallicacid.

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3.3. Determinationoftotalﬂavonoids

Totalflavonoidswereestimatedusingthemethodof Ordon-ezetal.[19].Inbrief,halfamlof2%AlCl3ethanolic solutionwasaddedto0.5mloftheextracts.After1hat roomtemperature,theabsorbancewasreadat420nm.The developmentofyellowcolorwastakingasindicationof thepresenceofflavonoids.Extractsampleswereevaluated atfinalconcentrationof1mg/ml.Totalflavonoidcontent wascalculatedasquercetinequivalent(mg/g)usingthe equationobtainedfromthecalibrationcurve.

3.4. Ulcerinduction

Gastriculcerationwasinducedintheanimalsaccording totheproceduredescribedbySayantietal.[20].Brieﬂy,rats wereadministeredwithasingleoraldoseofindomethacin (30mg/kgbodyweight).Theyweredeprivedoffoodbut had free access to water 24hprior to ulcer induction. Variousdegrees of ulceration have manifested 4hafter indomethacinadministration.

3.5. Animalgroupingandtreatments

Sixty three albino rats were randomized into nine groups of seven rats each. Group 1 (normal control) animals received only distilled water. Rats in group 2 (ulceratedcontrol)weregivenonlyindomethacin.Groups 3 and 4 animals were administered respectively with 200mg/kgb.w. of FE and SM only (to monitor likely negative/toxicological effect of the extracts). Animals in group 5 were given indomethacin after pretreat-ment with esomeprazole (20mg/kgb.w.). Groups 6, 7, 8 and 9 comprised ulcerated rats pretreated with FE (100mg/kgb.w.),FE(200mg/kgb.w.),SM(100mg/kgb.w.) andSM(200mg/kgb.w.)respectively.Treatmentswiththe reference drug and extractslasted for 21 days prior to indomethacinadministration. Thesewere orally admin-istered once daily using oral intubator with adlibitum provisionoffoodandwaterthroughouttheexperimental period.

3.6. Isolationofstomachandcollectionofgastricjuice Onthetwenty-thirdday(4hpostulcerinduction),the animalswerehumanelysacriﬁcedbycervicaldislocation. Theabdomenwasopenedandthestomachexcised.The stomachwasthereafter openedalong greatercurvature andgastriccontentwasdrainedintoacentrifugetube.Five mlofdistilledwaterwasaddedandtheresultantsolution wascentrifugedat3000rpmfor10min.Thesupernatant obtained was thereafter used for biochemical analyses. Thecleanedstomachswerepreservedin0.1Mphosphate salinebuffer(1:4(w/v),pH7.4)priortomacroscopic exam-inationandhomogenization.

3.7. Determinationofgastricsecretionparameters

Gastric acidoutput(volume) was determinedin the supernatant(2ml)bytitrationwith0.0025NNaOHusing Toepfer’s reagent as indicator. The pH of gastric juice

Table1

Ulcerscoresanddescriptiveremark.

Score Remark

0 Almostnormalmucosa

1 Vascularcongestions

2 Oneortwolesions

3 Severelesions

4 Veryseverelesions

5 Mucosafulloflesions

wasdeterminedusingapHmeter,whiletheprocedures

ofSanyal et al. [21]and Corneet al.[22] wereused to

determinespeciﬁcpepsinactivityandmucinconcentration respectively.

3.8. Quantiﬁcationofulceration

Degreesofulcerationintheindomethacin-treated ani-mals were quantiﬁed using the procedure outlined by SzaboandHollander[23].Brieﬂy,cleanedstomachswere pinnedonacorkboardandulcerswerescoredusing dis-sectingmicroscope withsquare-grid eyepiece based on gradingona0–5scale(depictingseverityofvascular con-gestionsandlesions/hemorrhagicerosions)aspresentedin Table1.Areasofmucosaldamagewereexpressedasa per-centageofthetotalsurfaceareaoftheglandularstomach estimatedinsquaremillimeters.Meanulcerscoreforeach animalwasexpressedasulcerindex(U.I)andthe percent-ageofinhibitionagainstulcerationwasdeterminedusing theexpressions:

U.I.=[Ulceratedarea/totalstomacharea]×100. %Ulcerinhibition=[U.I.incontrol−U.I.intest]

×100/U.I.incontrol.

3.9. Preparationofstomachhomogenateandassayof antioxidantindices

Immediatelyafterulcerscoring,wholestomachtissues weregroundwithliquidnitrogeninamortar.Theground tissues (0.5geach) were then homogenizedin ice cold 0.1Mphosphatesalinebuffer(1:4(w/v),pH7.4)andthe homogenatescentrifugedat2500rpmfor10minat4◦C. Theresultingsupernatantswerefrozenat₋₂₀◦Ctoensure maximum release of theenzymes located in thetissue beforebeingusedfortheenzymeassay.

Activity of superoxide dismutase (SOD) and level of lipid peroxidation measured in terms of malondialde-hyde (MDA) were respectively assayed in the stomach homogenatebythemethodsofMarklundandMarklund [24]andDevasagayamandTarachand[25].

3.10. Statisticalanalysis

Inhibition against ulceration was expressed in per-centage.Otherresultswereexpressedasmeanofseven determinations±standarderrorofmean.One-way anal-ysisofvariance (ANOVA) complementedwithStudent’s

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Table2

TotalphenolicandﬂavonoidcontentsofaqueousleafextractsofS.mombin andF.exasperata.Valuesexpressedpergofplantextract.

Aqueous extract Totalphenol(mg gallicacidg−1₎ Totalﬂavonoids (mgquerceting−1₎ S.mombin 85.50±0.20 60.53±0.10 F.exasperata 68.40±0.15 42.63±0.20

Valueswereexpressedpergofplantextractandaremeansoftriplicate

determination±standarddeviation.

t-testusingSPSSsoftwarepackageforwindows(Version

16) for differences between means was used to detect

anysigniﬁcantdifference(p<0.05)betweenthetreatment

groupsinthisstudy.

4. Results

Quantitative phytochemical analysis of aqueous leaf

extractsofSMandFErevealedthepresenceoftotalphenols

andﬂavonoids(Table2).

TheeffectsofaqueousleafextractsofSMandFEonthe ulcerindexand%inhibitionagainstulcerinthe experimen-talanimalsareshowninFigs.1and2respectively.Oral administrationof 30mg/kgb.w.of indomethacincaused a signiﬁcant(p<0.05) increase in thedegree of ulcera-tion(ulcerindex)intherats.Asigniﬁcantimprovement inthelevelofinhibitionagainstulcerationwashowever observedintheextracts-treatedanimals.Theextractsat 200mg/kgb.w.offeredbetterprotectionagainstulceration

Fig.1.Effect ofaqueous leafextracts ofS. mombin andF. exasper-ataonulcerindexofindomethacinulceratedrats(n=7,X±SEM).Bars withdifferentsuperscriptsfortheparameteraresigniﬁcantly differ-ent(p<0.05).IND:indomethacin(30mg/kgb.w.),ESP: esomeprazole (20mg/kgb.w.),FE1:Ficusexasperata(100mg/kgb.w.),FE2:Ficus exas-perata(200mg/kgb.w.),SM1:Spondiasmombin(100mg/kgb.w.),SM2: Spondiasmombin(200mg/kgb.w.). 0 10 20 30 40 50 60 70 80 90 100 IND (Ulcerated control)

IND + ESP IND + FE1 IND + FE2 IND + SM1 IND + SM2

% U lce r inh ibit ion

Fig.2. EffectofaqueousleafextractsofS.mombin andF.exasperata ondegreeofprotectionagainstulcerationinindomethacinulcerated rats (n=7, X±SEM). Bars represent % degree of protection offered againstulceration.IND:indomethacin(30mg/kgb.w.),ESP:esomeprazole (20mg/kgb.w.),FE1:Ficusexasperata(100mg/kgb.w.),FE2:Ficus exas-perata(200mg/kgb.w.),SM1:Spondiasmombin(100mg/kgb.w.),SM2: Spondiasmombin(200mg/kgb.w.).

Fig.3. Effectsofaqueous leafextractsofS. mombinandF. exasper-ata ongastricvolume andpHofindomethacinulceratedrats (n=7, X±SEM).Barswithdifferentsuperscriptsforeachparameterare sig-niﬁcantlydifferent(p<0.05).IND:indomethacin(30mg/kgb.w.),ESP: esomeprazole(20mg/kgb.w.),FE1:F.exasperata(100mg/kgb.w.),FE2: F.exasperata(200mg/kgb.w.),SM1:S.mombin(100mg/kgb.w.),SM2:S. mombin(200mg/kgb.w.).

thanthe100mg/kgb.w.regimensandcomparedwellwith thestandarddrug(Esomeprazole)used.

Fig.3showstheeffectofaqueousleafextractsofSM andFEongastricsecretionsofindomethacinulceratedrats. Indomethacinadministrationcausedsignificant(p<0.05) decrease in pH value with a corresponding significant (p<0.05)increaseingastricvolumeofgastriccontent. Pre-treatmentwiththeextractsproducedsignificantincrease in pHvaluecoupledwithsignificantdecreasein gastric volumewhencomparedwithulceratedcontrolrats.

Indomethacinadministrationbroughtabouta signifi-cant(p<0.05)increaseinspecificpepsinactivityaswell assignificantreduction(p<0.05)inmucincontentof gas-tricjuiceofulceratedratswhencomparedwiththenormal control(Fig.4).Theobservedchangesintheseparameters weresignificantlyattenuated(p<0.05)intheSMandFE extracts-treated rats.TreatmentwithSM revealedmore potentefficacyinthemodulationofbothpepsinandmucin contentsofgastricjuiceofulceratedrats.

Fig.4. EffectsofaqueousleafextractsofS.mombinandF.exasperata onpepsinactivityandmucincontentofindomethacinulceratedrats (n=7,X±SEM).Barswithdifferentsuperscriptsforeachparameterare signiﬁcantlydifferent(p<0.05).IND:indomethacin(30mg/kgb.w.),ESP: esomeprazole(20mg/kgb.w.),FE1:F.exasperata(100mg/kgb.w.),FE2: F.exasperata(200mg/kgb.w.),SM1:S.mombin(100mg/kgb.w.),SM2:S. mombin(200mg/kgb.w.).

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Fig. 5.Effect of aqueous leaf extracts of S. mombin and F. exas-perata on gastric. Malondialdehyde (MDA) level of indomethacin ulcerated rats (n=7, X±SEM). Bars with different superscripts for theparameteraresigniﬁcantlydifferent(p<0.05).IND:indomethacin (30mg/kgb.w.),ESP:esomeprazole(20mg/kgb.w.),FE1:Ficusexasperata (100mg/kgb.w.),FE2:Ficusexasperata(200mg/kgb.w.),SM1:Spondias mombin(100mg/kgb.w.),SM2:Spondiasmombin(200mg/kgb.w.).

Figs.5and6revealedtheeffectsofaqueousleafextracts ofSMandFEonthelipidperoxidationandSODactivity of gastricmucosalofindomethacinulceratedrats.MDA levelwassignificantlyincreased(p<0.05)intheulcerated animals(Fig.1).Asignificantreduction(p<0.05)wasalso observedintheactivityofSOD(Fig.2)inthe indomethacin-inducedanimals.Commendably,bothextractsparticularly at200mg/kgb.w.regimenresultedinsignificant improve-ment (p<0.05)in these parameters and the observable effectscomparedfavorablywellwithbothnormalcontrol andstandarddrug(esomeprazole)employedinthestudy. 5. Discussion

Inhibitoryactionofindomethacinonprostaglandin syn-thesiscoupledwithfreeradicalsformationhasbeenopined ascriticalbiochemicaleventsinthepathogenesisof gas-triculceration[26–28].Anunderstandingoftheseevents mightbeofutmostrelevanceindesigningnewantiulcer drugs. With the inherent adverse side effects and con-siderablyhighcostofsyntheticdrugs,exploitingnatural productsofplantsourcewhicharebelievedtobenon-toxic, efﬁcaciousandaffordablewillbemostappropriateinthe treatmentofgastriculcer.Phytotherapyisrapidlygaining groundsinsustaininghumanhealthandintheprevention

Fig. 6.Effect ofaqueous leafextractsof S.mombin and F. exasper-ata ongastricsuperoxidedismutase (SOD)activity ofindomethacin ulcerated rats (n=7, X±SEM). Bars with different superscripts for theparameteraresigniﬁcantlydifferent(p<0.05).IND:indomethacin (30mg/kgb.w.),ESP:esomeprazole(20mg/kgb.w.),FE1:Ficusexasperata (100mg/kgb.w.),FE2:Ficusexasperata(200mg/kgb.w.),SM1:Spondias mombin(100mg/kgb.w.),SM2:Spondiasmombin(200mg/kgb.w.).

ofcertaindiseaseslikegastriculcerresultingfromdrug toxicity[28,29].Thishasbeenascribedtopossessionof phytonutrientswithexcellentantioxidantpropertiesthat playsignificantrolesin managingtoxicityrelated disor-ders.Interestingly,studieshaverevealedthepresenceof someof thesebioactiveprinciplesinSMand FEaswell asreportedthemtopromotegoodhealth[5,8,14,7].Inthis study,wehavealsospecificallyquantifiedthepolyphenolic constituentsofaqueousleafextractsofSMandFEaswellas comparedtheirgastroprotectiveeffectson indomethacin-inducedulcerationinrats.

Biochemicalanalysisofgastricsecretions(forpH, gas-tric volume, bicarbonate,pepsin) and mucosalintegrity for stomach is usually employed to ascertainits status following exposuretopharmacological agents [30]. The pHgives an idea of thelevel of acidityand volume of gastric secretions. Low pH value is a manifestation of decreasedhydrogenionconcentrationingastricjuice.This hasbeenlinkedtopathogenesisofulcerandgastric dam-ageinexperimentalanimals[31].Inasetal.[27]havealso attributedgastrointestinalinjurytoerodedmucincontent. This erosion is facilitated by onslaughts of both inter-nal(pepsinandoxidantsproducedinthegastriclumen) andexternal(drugsandchemicals)aggressiveagentson mucosalepithelia.

Inthepresentstudy,thesigniﬁcantincrease inulcer index and gastric volume following oral administra-tion of indomethacin in the ulcerated rats may be attributed to either free radicals formation or inhibi-tionofprostaglandinsynthesis. Decreasedprostaglandin level has been attributed to impaired gastroprotection andincreasedgastricacidsecretionwhichareimportant eventsintheetiologyofmucosalulceration.Thisagrees withthereportsofBechetal.[32],Biplabetal.[30]and Muhammedetal.[33]whereindomethacinwasreported tohavecausedalterationsingastricsecretionsofrats. Con-versely,pretreatmentswiththetwoextractssigniﬁcantly reducedtheseparameters.Infacttheeffects noticedfor pHcomparedfavorablywellwithbothnormalcontroland standarddrugusedinthisstudyandindeedsuggestiveof theirpossiblegastroprotectiveattributes.

A combination of events including release of pre-formedmucus,woundretractionandre-epithelializationis involvedinulcer-healingprocessaftertoxicologicalinjury [23,34].Besidesprovidingsigniﬁcantbufferingcapacityfor theneutralizationofluminalacid,themucusalsooffers protectionagainstbothendogenousaggressorsand exoge-nous gastrotoxic agents such as indomethacin, thereby enhancingtherateoflocal healingprocess [35].In this study,theincreasedpepsinactivitycoupledwithdecrease in mucin secretion in the indomethacin-ulcerated rats indicatedalteredhydrophobicityandreducedprotective ability of the mucosal membrane against hemorrhagic erosion, thus, resulting in tissue damage. This implied decreasedabilityofthegastricmucosatowithstandthe offensiveonslaughtofindomethacin.Besidesantioxidant actionthatprotectsthemucuslayerandarrestsulcer pro-gression,drugsthatincreasethesynthesisandsecretion ofgastric mucuswould accelerate gastriculcerhealing. Pretreatmentwiththeextractshowever,facilitatedulcer healingprocess,whichisassociatedwithdecreasedpepsin

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activityand elevatedmucinlevelinthegastricmucosa. This in turn has encouraged speedy wound healing of theulceratedareasofthemucosalepitheliaandshielded thegastrointestinalmembrane,thus abrogatingthe cat-astrophicinﬂuenceofindomethacinintheulceratedrats [34].Thisisindicativeofenhancedmucussecretory poten-tialoftheextractsandsuggestiveoftheirsigniﬁcantrolein ulcerhealingprocess.Healingofmucosaepitheliacellswas prominentlydisplayedbytheextractsat200mg/kgb.w. dose,depictinga betterulcerhealingcapacity and com-paredfavorablywellwiththereferencedrugused.

Cells or tissues are in a stable state if the rates of freeradicalformationandscavengingcapacityare essen-tiallyconstantandinequilibrium.However,animbalance betweenthem resultsin oxidative stress which further deregulatescellularfunctionsleadingtodifferent patho-logicalconditions[17].Inthepresentstudy,theincreased concentrationofMDAaswellasreducedactivityofSODin thestomachofindomethacin-ulceratedratsisa manifes-tationoffacilitatedlipidperoxidationandoverproduction offree radicals resultingin mucosaldamage. Free radi-calsthwartantioxidantenzymesactivitiesandinitiatelipid peroxidationwhichisanimportanteventinthetoxicity mechanismofindomethacin[36].Indomethacinhas previ-ouslybeenreportedtodecreaseantioxidantenzymes(SOD, CATandGST)activityinratstomachtherebyinducing gas-triculceration[37].Thisisassociatedwithoverpowering ofthecellularantioxidantdefensesystemsbyfreeradicals ravaginginﬂuence thatsubsequentlyresultsin stomach oxidativeinjury.However,thesigniﬁcantlyreduced con-centrationsofMDAcoupledwithmarkedincreaseinthe activityofSODinratspretreatedwithaqueousleafextracts ofbothplantsisanobviousindicationofantiperoxidative potentialandthusantioxidativepotential.

Generally,theprotectionofferedbytheaqueousleaf extractsofSMandFEagainstindomethacin-induced gas-triculcerationmaybelinkedtotheirbeneﬁcialmedicinal attributes occasioned by phytometabolite constituents. Theseincludeabilitytoscavengefreeradicalsand regu-latemucosalmembranepermeabilitytherebycountering theeffectofindomethacinongastricacidsecretion.This isinagreementwiththesubmissions ofInasetal.[27], Muhammed et al. [33] and Gege-Adebayo et al. [38], wheregastroprotectivepotentialsofplantextractsagainst indomethacin-ulceratedrats wereassociated with their polyphenoliccompoundsandothervariousbioactive prin-ciples.Sinceesomeprazoleisaprotonpumpinhibitor,then theeffectproducedbythetwoextractsmighthaveperhaps mimicitsmechanismofactionbymodulatingcellsinthe mucosalliningofthestomachagainstexcessacidsecretion [39,40].

6. Conclusion

Overall, the attenuation of gastric affronts of indomethacinbyadministrationofaqueousleafextracts ofSMand FEat 200mg/kgb.w.regimenis indicativeof theirexcellentgastroprotectiveandantioxidative poten-tialsinrats.Effortsareongoingtoinvestigatetheexact antiulcerogenic principle(s) in these extracts and also

harnesstheirpossiblesynergisticefﬁcacyagainstgastric ulcer.

Conﬂictofinterest

The authors declared that there are no conﬂicts of interest.

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