@@@@@_____Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity☆ _...
-
Upload
aurelioarae -
Category
Documents
-
view
214 -
download
0
Transcript of @@@@@_____Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity☆ _...
-
8/9/2019 @@@@@_____Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity _ Julio Tobn - Aca
1/9
17/4/2014 Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity| Julio Tobn - Academia.edu
http://www.academia.edu/4712295/Curcumin_restores_Nrf2_levels_and_prevents_quinol inic_acid- induced_neurotoxici ty_ 1/9
Home
Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicityby Julio Tobn
more
CurcuminrestoresNrf2levelsandpreventsquinolinicacid-inducedneurotoxicity
IvnCarmona-Ramreza,AbelSantamarab,JulioC.Tobn-Velascob, c,MarisolOrozco-Ibarra d,IrmaG.Gonzlez-Herrerab,JosPedraza-Chaverr c,PerlaD.Maldonadoa,
aPatologaVascularCerebral,InstitutoNacionaldeNeurologayNeurociruga ManuelVelascoSurez,MxicoD.F.,14269,MxicobAminocidosExcitadores,InstitutoNacionald eNeurologay Neurociruga ManuelVelascoSurez,MxicoD.F.,14269,Mxico
cDepartamentodeBiologa,FacultaddeQumica,UniversidadNacionalAutnomadeMxico,MxicoD.F.,04510,MxicodNeurobiologaMolecularyCelular,INNN-UNAM,InstitutoNacionaldeNeurologayNeurociruga ManuelVelascoSurez,MxicoD.F.,14269,Mxico
Received5July2011;receivedinrevisedform13December2011;accepted22December2011
Abstract
Neurologicaldiseasescompriseagroupofheterogeneousdisorderscharacterizedbyprogressivebraindysfunctionandcelldeath.Inthenextyears,these
diseasesareexpectedtoconstituteaworld-widehealthproblem.Becauseexcitotoxicityandoxidativestressareinvolvedinneurodegenerativediseases,it
becomesrelevantto describepharmacological therapies designed to activate endogenouscytoprotective systems. Activationof transcription factor Nrf2
stimulatescytoprotectivevitagenesinvolvedinantioxidantdefense.Inthiswork,weinvestigatedtheabilityoftheantioxidantcurcumintoinducetranscription
factorNrf2inaneurodegenerativemodelinducedbyquinolinicacidinrats.Animalswereadministeredwithcurcumin(400mg/kg, p.o.)for10days,andthen
intrastriatallyinfusedwithquinolinicacid(240nmol)onday10oftreatment.Curcuminpreventedrotationbehavior(6dayspost-lesion),striatalmorphological
alterations(7 dayspost-lesion)and neurodegeneration(1 and3 dayspost-lesion)i nducedbyquinolinicacid.Curcuminalso reducedquinolinicacid-inducedoxidativestress(measuredasproteincarbonylcontent)at6h post-lesion.Theprotectiveeffectsofcurcuminwereassociatedtoitsabilitytopreventthe
quinolinicacid-induceddecreaseofstriatalintra-nuclearNrf2levels(30and120minpost-lesion),andtotalsuperoxidedismutaseandglutathioneperoxidase
activities(1daypost-lesion).Therefore,resultsofthisstudysupporttheconceptthatneuroprotectioninducedbycurcuminisassociatedwithitsabilityto
activatetheNrf2 cytoprotectivepathwayandto increasethetotal superoxidedismutaseandgluta thioneperoxidaseactivities.
2013ElsevierInc.Allrightsreserved.
Keywords: Nrf2;Curcumin;Quinolinicacid;Oxidativestress
1.Introduction
Neurodegenerationistheresultofpathologicalprocessesproduc-
ingsevereandspecificpatternsofbraincelldamageinaconcertedmanner [1,2].Neurodegenerativeeventsconstituteamajorcausefor
development of neurological disorders. Humandiseases coursingwithneurodegenerationinvolveexcitotoxicityasatriggeringevent
fortheinitiationofdeadlycascades [3-5].Excitotoxicityiscurrentlydefinedasatoxicprocesscharacterizedbyasustainedstimulationof
excitatoryaminoacidreceptors [6-8],mainlyinvolvingN-methyl-D-aspartatereceptors(NMDAr).Differenttoxiceventsderivedfrom
excitotoxicityhavebeencharacterizedinexperimentalmodels,and
they include upregulation of detrimental signalingpathways,dis-
ruptedCa2+ homeostasis,andrecruitmentofreactiveoxygenand
nitrogenspecies(ROS/RNS),withfurtheroxidative/nitrosativestress
[2,8-11],ultimatelyleadingtocelldeath.Quinolinic acid (QUIN) is a glutamatergic agonist acting on
NMDAr, preferentially in discrete populations of these receptorscontaining the NR2A and NR2B subunits. This metabolite is
synthesized in the kynurenine pathway, and is normallypresentat nanomolar concentrations in human and rat brains [12].
However, under pathological conditions, kynurenine pathway isstimulatedto increasethe levels of QUIN, thereforeaugmenting
therisk forexcitotoxic events.Then, thistoxicmetabolite exertsexcessiveexcitationofNMDArandrecruitsenhancedcytoplasmic
Ca2+ concentrations, mitochondrial dysfunction, decreased ATP
levels,cytochromecreleaseandoxidativestress,furtherleadingto
selectiveloss of GABAergic and cholinergic neurons [13]. Indeed,wheninjectedintothebrain,QUINreproducesneurodegenerative
events in rodents, resembling those observed in the brains of
patients with Huntington's disease [14]. In addition, the intras-
triatalinfusionofQUINtorodentsstimulateslipidperoxidationin
thisregionat shorttimes [15],andthese findingsarerelatedto
increased extracellular levels of hydroxyl radical (OH) in the
samebrainregion [16].Furthermore,QUINisknowntogeneratea
dysregulation in the oxidant/antioxidant ratio by affecting the
Available online at www.sciencedirect.com
JournalofNutritionalBiochemistry24(2013)14 24
ThisworkwassupportedbyCONACYT(GrantNo.103527toPDMand
129838toJPCH)andbyPAPITIN121910. Correspondingauthor. PatologaVascular Cerebral,InstitutoNacional
deNeurologayNeurocirugaManuelVelascoSurez,InsurgentesSur3877,
MxicoD.F.,14269,Mxico.Tel.:+525556063822x2009;fax:+52555424
0808.
E-mailaddress: [email protected](P.D.Maldonado).
0955-2863/$-seefrontmatter2013ElsevierInc.Allrightsreserved.
http://dx.doi.org/10.1016/j.jnutbio.2011.12.010
reducedglutathione:oxidizedglutathione(GSH:GSSG)ratio,aswell
a s depleting the activity of Cu,Zn-SOD at different post-lesion
times [17], alsorecruitingthe early andtime-dependent formation
of peroxynitrite (ONOO-) as a key RNS contr ibuting to this
game,CA,USA).Allotherreagentswerepurchasedfromotherknowncommercial
sources.DeionizedwaterfromaMilli-QSystemofMillipore(Bedford,MA,USA)was
usedforthepreparationofallsolutions.
2.2.Animals
15I.Carmona-Ramrezetal./JournalofNutritionalBiochemistry24(2013)14 24
Search People, Research Interests and Universities
D ow nlo a
Curcumin_restore
1.74 MB
http://www.academia.edu/attachments/32037871/download_file?st=MTM5NzczOTk2OCwxNzcuNDMuNjUuMjQy&ct=MTM5NzczOTk3MQ%3D%3Dhttp://www.academia.edu/attachments/32037871/download_file?st=MTM5NzczOTk2OCwxNzcuNDMuNjUuMjQy&ct=MTM5NzczOTk3MQ%3D%3Dhttp://www.sciencedirect.com/science/journal/09552863http://dx.doi.org/10.1016/j.jnutbio.2011.12.010http://dx.doi.org/10.1016/j.jnutbio.2011.12.010http://dx.doi.org/10.1016/j.jnutbio.2011.12.010mailto:[email protected]://dx.doi.org/10.1016/j.jnutbio.2011.12.010http://dx.doi.org/10.1016/j.jnutbio.2011.12.010mailto:[email protected]://dx.doi.org/10.1016/j.jnutbio.2011.12.010http://dx.doi.org/10.1016/j.jnutbio.2011.12.010http://dx.doi.org/10.1016/j.jnutbio.2011.12.010http://www.sciencedirect.com/science/journal/09552863http://unam.academia.edu/JulioTob%C3%B3nhttp://www.academia.edu/4712295/Curcumin_restores_Nrf2_levels_and_prevents_quinolinic_acid-induced_neurotoxicity_http://www.academia.edu/http://www.academia.edu/ -
8/9/2019 @@@@@_____Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity _ Julio Tobn - Aca
2/9
17/4/2014 Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity| Julio Tobn - Academia.edu
http://www.academia.edu/4712295/Curcumin_restores_Nrf2_levels_and_prevents_quinol inic_acid- induced_neurotoxici ty_ 2/9
para gm . otewort y, t ese an o t er a t erat ons n uce
by QUIN can be prevented by different antioxidants such as
melatonin, sodium selenite, L-carnitine, epigallocatechin gallate,
etc. [15,19-21].Thisevidencehighlightsasubstantialcontributionof oxidative stress to the toxic pattern elici ted by QUIN.
Consequently,itcanbeassumedthatagentsexhibitingantioxidantproperties may have potential therapeutic valueat experimental
levelinthisandothermodels.Curcumin(CUR),thepolyphenolicnon-flavonoidyellowbioac-
tivecomponent of turmeric thepoweredrhizome of Curcuma
longa Linn ,hasbeenshowntoproduceawiderangeofpositive
biological effects through its antioxidant and anti-inflammatory
properties [22,23].CURisnon-toxicandhasbeenshowntobeapotent free radicalscavenger [24,25]. Inaddition,CUR, like other
polyphenols commonly found in plants, fruits and vegetables,
competes in efficacywith vitamins C and E [26], although recentfindingshaveattributedpartofitsanti-in flammatoryefficacytoits
morestablemetabolitetetrahydrocurcumin [27].Nevertheless,CUR
itself has been shown to cross the blood-brain barrier to exert
neuroprotective effects,suchas those reportedagainst homocyste-
ine-inducedcognitiveimpairment andoxidative stressin rats [25]. CUR
alsoinhibits amyloid toxicity in vivo [28] and improves memory
functionsinastreptozotocin-induceddementiamodelinratsthroughinsulin receptors [29]. One of the most relevant findingson CUR
researchis thedescriptionofits capacitytoupregulatethemaster
antioxidantcoordinator,transcriptionfactornuclear factorerythroid2-
related factor2 (Nrf2), whichin turn is responsible forphase 2
antioxidant and detoxification genes expression, such as heme
oxygenase-1,andthiseffectofCURisabletoprotecttheratbrain
fromfocalischemia [23].
DespiteafewgroupshavealreadydescribedprotectiveeffectsofCURon QUIN-inducedtoxicity includingantiperoxidativeactionsin rat brain homogenates [30], as well as antiexcitotoxic and
antioxidant effectsin primaryculturesof humanneurons [21] ,
to our knowledge there are no reports on the effects of thispolyphenoliccompoundinthetoxicmodelinducedbyQUINunder
invivoconditions,noronapossibleinvolvementofNrf2inCUR-induced neuroprotection. Therefore, theaim of thiswork wasto
evaluateifCURcanattenuateorpreventtheQUIN-inducedinvivotoxicity in rats, and whether this effect is dependenton Nrf2
upregulation.OurresultssupportaroleoftheNrf2pathwayintheneuroprotectiveactionsofCURinthisparadigm.
2.Methodsandmaterials
2.1.Reagentsandchemicals
CUR,QUIN,apomorphine,2,4-dinytrophenilhydrazine(DNPH),guanidinehydro-
chloride,4-(2-hydroxyethyl)-1-piperazineethanesulfonicacid (HEPES),sodium fluo-
ride(NaF), phenylmethylsulfonylfluoride,sodium pyrophosphate,sodium vanadate
(Na3VO4), ethylenediaminetetraacetic acid(EDTA), ethylene glycoltetraacetic acid
(EGTA),Nonidet P-40,glycerol, sodiumdodecylsulfate (SDS),leupeptin, glutathione
(GSH),glutathionereductase,NADPH,xanthine,xanthineoxidase,nitrobluetetrazo-
lium(NBT),andbovineserumalbumin(BSA),wereobtainedfromSigma-Aldrich(St.
Louis,MO,USA).Fluoro-jadeB(F-JB)wasobtainedfromMillipore(Bedford,MA,USA).
Anti-Nrf2antibodyusedforimmunofluorescence(C-20,sc-722)andWesternblot(sc-13032),anti-Lamin B1 antibody (sc-20682), and 4,6-diamidino-2-phenylindole
dihydrochloride (DAPI, sc-3598)were obtained from Santa CruzBiotechnology
(SantaCruz,CA,USA).Anti- -tubulinantibody(ab-11307)wasfromAbcam(San
Francisco,CA,USA). AlexaFluor488 goatanti-rabbitIgG secondaryantibodywas
obtained from Invitrogen (Carlsbad, CA,USA). Goat-antirabbit IgG horseradish
peroxidase-conjugate(62-6120)and goat-antimouseIgG horseradishperoxidase-
conjugate(sc-2005)secondaryantibodiesusedforWesternblotwerefromZymed
(San Francisco, CA, USA) and Santa Cruz Biotechnology(SantaCruz,CA, USA),
respectively.Vectashield mountingmedium wasobtained fromVector Labs(Burlin-
MaleWistarratsinitiallyweighing250 300g,wereusedthroughoutthestudy.
Forallexperimentalpurposes,animalswerehoused fivepercageinacrylicboxesand
providedwithstandardcommercialratchow(LabRodentDiet5001;PMIFeedsInc.,
Richmond,IN,USA)andwateradlibitum.Thehousingroomwasmaintainedunder
constantconditionsofhumidity(50%10%),temperature(25C3C)andlighting
(12 h dark-lightcycles). All procedures with animals were carried outstrictly
accordingto theNationalInstitutes ofHealth Guidelinesfor thecareand useof
laboratoryanimals andthelocalguidelinesontheethicaluseofanimalsfromthe
Health Ministry of Mexico. During the experiments, all efforts were made to
minimizeanimal suffering.
2.3.Experimentaldesign
All experiments were performedduringthe morning (starting every dayat
8:00AM).Theanimalswererandomlydividedintofourgroups,asmentionedbelow:
1)controlgroup(CT),treatedwithvehicleplussaline;2)CURgroup,treatedwithCUR
plussaline;3)QUINgroup,treatedwithvehicleplusQUIN;and4)CUR+QUINgroup,
treatedwithCURplusQUIN.AnimalsfromCURandCUR+QUINgroupsreceivedCUR(100,200or400mg/kg/day,p.o.)for10consecutivedays.CTandQUINgroupsreceived
vehiclefor10days.CURwasdissolvedincarboxymethylcellulose5%(vehicle).Onday
10,animalswereintrastriatallyinfusedwith1 lofQUINorsaline.QUIN-treatedrats
receivedasingleinfusionofQUIN(240nmol/L)intherightstriatum,accordingtothe
followingstereotaxiccoordinates:+0.5mmanteriortobregma,-2.6lateraltobregma,
and+4.5mmventraltothedura [31]. TheeffectsofincreaseddosesofCUR(100,200
and400mg/kg)oncirclingbehavior(6dayspost-lesion)andhistologicalalterations
(7days post-lesion)inducedby QUINwere firstevaluatedto select an optimum
protective doseof CUR.The mostprominent protective effect wasobservedwith
400mg/kg,sothisdosewasusedfromthispointonforfurtherassays.Neurodegenera-
tion(1and3dayspost-lesion),proteincarbonyllevels(6hpost-lesion),Nrf2activation
(30and120minpost-lesion),andactivitiesoftotalsuperoxidedismutase(SOD)and
glutathioneperoxidase(GPx)(1daypost-lesion)weredetermined.
2.4.Circling behavior
Motoralterations,assessedasrotationbehavior,wereevaluatedinanimalsfrom
allexperimentalgroups,accordingtopreviousreports [20,32].SixdaysafterQUIN
infusion,animalswereadministeredwithapomorphine(1mg/kg, s.c.)andseparatedintoindividualacrylicboxcages.Fiveminuteslater,thenumberofipsilateralrotations
tothelesionedsidewasrecordedfor1h.Eachrotationwasdefinedasacompleteturn
(360).Dataareexpressedasthetotalnumberofipsilateralturnsin1h.
2.5.Histologicalexamination
Braintissueswerecollectedandhistologicallyprocessedaccordingtoprevious
descriptions [20,32].Sevendaysafterintrastriatallylesioned,animalsfromallgroups
wereanesthetized i.p.with0.5mLofsodiumpentobarbitalandperfusedtranscardially
with 0.9%salinesolution containing heparin(200/1 v/v), followedby 4%p-
formaldehydeat4C.Brainswereremoved,post-fixedin4%p-formaldehydefor7
days andembedded in paraffin. Fixedtissues were serially sectionedin an 820
HistoSTAT microtome(AmericanInstrument Exchange, Inc.,Haverhill,MA, USA).
Striatalsections(5 m-thick)wereobtainedevery100 m,coveringatotaldistanceof
300 m(100 manteriorand100 mposteriortotheneedletract).Allsectionswere
stainedwith hematoxylin-eosin(H&E)to visualizecell bodies,usingan imageanalyzer
IM100(LeicaCambridge,UK).
2.6.Quantitativeassessmentof lesions
Cellcountingin histologically preparedsectionswas performedas previously
described [32].Thegeneralcriteriatoscoredamagedcellsincludedpyknoticnuclei,
cytoplasmic vacuolation, neuronal atrophy, interstitial edema and necrosis. The
numberof damagedand preservedneuronswas obtainedfrom 5randomly selected
fieldsinthreesectionslidesperbrain.Dataareexpressedaspercentageofdamaged
neuronsper field.
2.7.Detectionof neurodegenerationusingF-JBstaini ng
Braintissuewascollectedandprocessedaccordingtopreviousreports [33,34].
Oneand three days after intrastriatallylesioned,animals from all groups were
anesthetized i.p. with0.5 mLof sodium pentobarbital andtranscardiallyperfused
with 0.9%saline solution containingheparin (200/1v/v), followed by 4% p-
formaldehyde at4C. Brains wereremovedand post-fixedin 2%p-formaldehyde
and30%sucrosefor7daysat4C.Tissueswerethensubmergedinoptimalcutting
temperaturecompoundandfrozenoverliquidnitrogen,followedbycryosectionat
-25Cina cryostatmicromHM520(ThermoScientific,Pittsburg,PA,USA).Sections
(22 m-thick) were mounted onto glass slides coated with 0.5% gelatin and
immersedin 1%NaOHand80% ethanol for5 min, followedfor2 minin70%
ethanol.Slidesweretransferredto0.06%KMnO4 for10minonashakingtableto
assess consistent backgroundsuppression betweensections,and thenrinsed in
distilledwaterfor2min.Sectionswerestainedina0.0004%F-JBplus0.0002%DAPI
solutionfor20min.Slideswererinsedthricewithdistilledwaterfor1mineachstep.
Excesswaterwasremoved,placingthesectionsonaslidewarmerat50Cuntilfully
dry(510min),andthenclearedbyfurtherimmersioninxyleneforatleast1min
beforecoverslippingwithanon-aqueous,nonfluorescentplasticmountingmedia.
ImmunofluorescencewasvisualizedwithanimageanalyzerIM100(LeicaCambridge,
UK)usingagreen filterforF-JBandbluefi lterforDAPI.
2.8.Protein carbonylcontent
Asan indexofproteinoxidation,proteincarbonylcontentinthestriataltissue(6h
post-lesion)wasdeterminedaspreviouslydescribed [32].Assessmentofcarbonyl
formationwasdoneonthebasisofformationofproteinhydrazonebyreactionwith
DNPH.Striatalhomogenateswereincubatedwith10%streptomycinsulfatetoremove
nucleicacidsovernightandcentrifugedat21,000gat4Cfor20min.Further,striatal
homogenates were treated with 10mM DNPH (in2.5M HCl) for1 h atroom
temperature,and10%trichloraceticacidwasaddedandcentrifugedat2,500gat4Cfor10min.Pelletwaswashedthreetimeswithethanol/ethylacetate(1:1),dissolved
with 6 M guanidine hydrochloride (inphosphatebuffer 20 mM,pH 7.4), andcentrifugedat5,000gat4Cfor3mintoremoveinsolublematerial.Absorbancewas
measuredat370nm.ProteincarbonylcontentisexpressedasnmolDNPH/mgprotein,
usingthe molarabsorptioncoefficient ofDNPH(22,000 M-1 cm-1).Totalprotein
concentration wasobtainedbyreadingopticaldensityat 280nm inblanktubes
preparedin parallel(treatedonlywith2.5M HCl),usinga standardcurveofBSA(0.25
2mg/ml)preparedin6Mguanidinehydrochloride.
2.9.Detectiono Nr2 b immuno luorescence
KCl,1mMphenylmethylsulfonyl fluoride,20mMNaF,1mMsodiumpyrophosphate,
1mMNa3VO4,1%NonidetP-40,and1 g/mLleupeptin).Homogenateswerethen
centrifuged at 500g for5 min, thesupernatants were recovered as cytosolic
fractions,and thenuclear pellets were washedin cold bufferB (10mM HEPES
pH=8.0,0.1mMEDTA,1mMphenylmethylsulfonyl fluoride,20mMNaF,1mM
sodiumpyrophosphate, 1 mM Na3VO4, 25% glycerol,0.1 M NaCl, and 1 g/mL
leupeptin).Afterasecondsteptofcentrifugationat500gfor5min,thenucleiwere
resuspendedin2%SDShypertoniccoldbufferC(10mMHEPESpH=8.0,0.1mM
EDTA,1mMphenylmethylsulfonyl fluoride,20mMNaF,1mMsodiumpyrophos-
phate, 1 mMNa3VO4, 25% glycerol, 0.4 M NaCl, and 1 g/mL leupeptin),and
sonicated.Cytosolicandnuclearfractionswereresolved inSDS-PAGE(10%), using60 g
(cytoplasmicfraction) or 80 g (nuclear fraction) protein/lane,transferred into
Immobilon-P membranes (PVDF, Millipore), and immunoblotted with primary
antibodies:anti-Nrf2,anti--tubulinandanti-LaminB1(1:1,000).Peroxidase-conju-
gatedsecondaryantibodies(1:10,000)wereusedtodetecttheproteinsofinterestby
enhancedchemiluminescence,accordingtoapreviousreport [35]. Thecorresponding
bandswerequantifiedbydensitometry.
2.11.SOD activity
TotalSODactivity(MnSODandCuZnSOD)inhomogenateswasdeterminedusing
xanthine-xanthineoxidasesystemtoreduceNBT.Mixturereactioncontainedinafinalconcentration:0.122mMEDTA,30.6 MNBT,0.122mMxanthine,0.006%BSA,and
49mMsodiumcarbonate.Fivehundred lofhomogenates(1:20)wereaddedto2.45 mlof themixture describedabove, then 50 l xanthine oxidase, in a final
concentrationof2.8U/l,wereaddedandincubatedinawaterbathat27Cfor30min.
Thereactionwasstoppedwith1mlof0.8mMcupricchlorideandtheopticaldensity
wasreadat560nm.OnehundredpercentofNBTreductionwasobtainedinatubein
whichthesamplewasreplacedby distilledwater.T heamountof proteinthatinhibited
NBTreductionto50%ofmaximumwasdefinedasoneunitofSODactivity.Results
wereexpressedasU/mgprotein.
16 I.Carmona-Ramrezetal./JournalofNutritionalBiochemistry24(2013)14 24
-
8/9/2019 @@@@@_____Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity _ Julio Tobn - Aca
3/9
-
8/9/2019 @@@@@_____Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity _ Julio Tobn - Aca
4/9
17/4/2014 Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity| Julio Tobn - Academia.edu
http://www.academia.edu/4712295/Curcumin_restores_Nrf2_levels_and_prevents_quinol inic_acid- induced_neurotoxici ty_ 4/9
-
8/9/2019 @@@@@_____Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity _ Julio Tobn - Aca
5/9
17/4/2014 Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity| Julio Tobn - Academia.edu
http://www.academia.edu/4712295/Curcumin_restores_Nrf2_levels_and_prevents_quinol inic_acid- induced_neurotoxici ty_ 5/9
-
8/9/2019 @@@@@_____Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity _ Julio Tobn - Aca
6/9
17/4/2014 Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity| Julio Tobn - Academia.edu
http://www.academia.edu/4712295/Curcumin_restores_Nrf2_levels_and_prevents_quinol inic_acid- induced_neurotoxici ty_ 6/9
-
8/9/2019 @@@@@_____Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity _ Julio Tobn - Aca
7/9
17/4/2014 Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity| Julio Tobn - Academia.edu
http://www.academia.edu/4712295/Curcumin_restores_Nrf2_levels_and_prevents_quinol inic_acid- induced_neurotoxici ty_ 7/9
-
8/9/2019 @@@@@_____Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity _ Julio Tobn - Aca
8/9
17/4/2014 Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity| Julio Tobn - Academia.edu
http://www.academia.edu/4712295/Curcumin_restores_Nrf2_levels_and_prevents_quinol inic_acid- induced_neurotoxici ty_ 8/9
-
8/9/2019 @@@@@_____Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity _ Julio Tobn - Aca
9/9
17/4/2014 Curcumin restores Nrf2 levels and prevents quinolinic acid-induced neurotoxicity| Julio Tobn - Academia.edu
http://www academiaedu/4712295/Curcumin restores Nrf2 levels and prevents quinol inic acid induced neurotoxici ty 9/9
Job Board About Mission Press Blog Stories We're hiring engineers! FAQ Terms Privacy Copyright Send us Feedback
Academia 2014
http://www.academia.edu/feedbackhttp://www.academia.edu/copyrighthttp://www.academia.edu/privacyhttp://www.academia.edu/termshttp://www.academia.edu/FAQhttp://www.academia.edu/hiringhttp://www.academia.edu/storieshttp://blog.academia.edu/http://www.academia.edu/presshttp://www.academia.edu/hiring/missionhttp://www.academia.edu/abouthttp://www.academia.edu/Jobs