Antidepressant-Like and Antioxidant Effects of Plinia trunciflora in Mice

Research ArticleAntidepressant-Like and Antioxidant Effects ofPlinia trunciflora in Mice

Cassia Sacchet1 Ricieri Mocelin1 Adrieli Sachett2 Fernanda Bevilaqua2 Rafael Chitolina2

Fernanda Kuhn1 Aline Augusti Boligon3 Margareth Linde Athayde3

Walter Antonio Roman Junior2 Denis Broock Rosemberg14 Jacir Dal Magro1

Greicy Michelle Marafiga Conterato15 and Angelo L Piato16

1Programa de Pos-Graduacao em Ciencias Ambientais Unochapeco Avenida Senador Attılio Fontana 591E89809-000 Chapeco SC Brazil2Nucleo de Fitoterapicos Programa de Pos-Graduacao em Ciencias da Saude Unochapeco Avenida Senador Attılio Fontana 591E89809-000 Chapeco SC Brazil3Laboratorio de Fitoquımica Universidade Federal de Santa Maria Avenida Roraima 1000 97105-900 Santa Maria RS Brazil4Programa de Pos-Graduacao em Bioquımica Toxicologica Universidade Federal de Santa Maria Avenida Roraima 100097105-900 Santa Maria RS Brazil5Laboratorio de Fisiologia da Reproducao Animal Universidade Federal de Santa Catarina Rodovia Ulisses Gaboardi Km 3Campus Curitibanos 89520-000 Curitibanos SC Brazil6Programa de Pos-Graduacao em Farmacologia e Terapeutica Universidade Federal do Rio Grande do SulAvenida Sarmento Leite 500305 90050-170 Porto Alegre RS Brazil

Correspondence should be addressed to Angelo L Piato angelopiatogmailcom

Received 10 March 2015 Revised 2 June 2015 Accepted 11 June 2015

Academic Editor Menaka C Thounaojam

Copyright copy 2015 Cassia Sacchet et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

The jaboticaba tree Plinia trunciflora (O Berg) Kausel is popularly named ldquojabuticabeirardquo in Brazil and is used in folk medicine totreat diabetes and chronic inflammation of the tonsils but studies evaluating the central effects of this species are limitedThis studyevaluated the antidepressant-like and antioxidant effects of P trunciflora (PT) aqueous extract in which five different anthocyaninswere identified PT showed significant ferric-reduction power and DPPH radical scavenging activity in vitro and reduced lipidperoxidation both in vitro and ex vivo At the behavioural level PT (400 and 800mgkg ip) dose-dependently reduced immobilitytime in the tail suspension test in Swiss male mice The identification of bioactive compounds accompanied by the in vitro and exvivo antioxidant activity of PT suggests that these activities might be related to the antidepressant-like activity of P trunciflora

1 Introduction

Depression is a common serious and recurrent chronicaffective disorder characterized by anhedonia headachesleep disturbances changes in sexual desire and a loss ofenergy [1] This disease is among the five most prevalent inthe world and is expected to be the second leading causeof disability in 2020 [2] The monoaminergic hypothesisof depression [3] does not provide a full understanding ofthe progression causes and pharmacotherapy of depressionNew hypotheses have been postulated and oxidative stress

has been suggested to be involved in the pathophysiology ofdepression [4]

Oxidative stress is a condition in which an imbalancebetween the production of free radicals and endogenousantioxidant defenses occurs [5] culminating in decreased cellantioxidant capacityThe superoxide anion (O2

∙minus) and hydro-gen peroxide (H

2

O2

) produced during respiratory chain maygenerate the highly deleterious hydroxyl radical (∙OH) viathe Fenton reaction [6] The overproduction of these speciesis related to protein DNA and lipid oxidation [7] as wellas the inactivation of important antioxidant enzymes such

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2015 Article ID 601503 9 pageshttpdxdoiorg1011552015601503

2 Evidence-Based Complementary and Alternative Medicine

as catalase (CAT) superoxide dismutase (SOD) glutathioneperoxidase (GPx) and thioredoxin reductase (TrxR) [5]The overproduction of ROS and oxidative stress have beenimplicated in the pathophysiological processes related tovarious diseases including Alzheimerrsquos Parkinsonrsquos anx-iety and depression [8ndash10] In this sense plants emergeas potential alternatives for the treatment of oxidativestress-related diseases considering that they are importantsources of carotenoids flavonoids vitamins and polyphe-nols

TheMyrtaceae family consists of 4620 species distributedin 140 genera whose occurrence has been described insubtropical and tropical regions of the world mainly Cen-tral and South America and Australia [11] The jaboticabatree Plinia trunciflora (O Berg) Kausel a synonym ofMyrciaria trunciflora O Berg Eugenia cauliflora O Bergand Myrciaria peruviana (Poir) Mattos is popularly namedldquojabuticabeirardquo in Brazil (source httpwwwtropicosorg)In folk medicine species of Plinia have been used to treatvarious diseases such as diabetes and chronic inflammationof the tonsils [12] However studies that evaluate the effects ofthis species on the central nervous system (CNS) are scarce inthe literature Therefore the aim of this study was to evaluatethe antidepressant-like effect of P trunciflora aqueous extractin the tail suspension test The identification of bioactivecompounds and the in vitro and ex vivo antioxidant effectsof P trunciflora were investigated in order to establish if theantidepressant-like effect of this plant could be related tothese properties

2 Materials and Methods

21 Plant Material The whole fruits of Plinia trunciflorawere collected in Alpestre (RS Brazil) (27∘101015840568210158401015840S and53∘71015840195510158401015840O) in September and taxonomically identified byMarcos Eduardo Guerra Sobral (botanical) where a voucherhas been deposited in the university herbarium (number3302)

22 Preparation of Aqueous Extract of P trunciflora (PT)The aqueous extracts of whole fruits were prepared basedon the methodology described by Kuskoski et al [13] Thewhole fruit (100 g) of P trunciflora was mixed with 200mLof distilled water and acidified with concentrated HCl untilpH 15 After trituration for 1 min the solution was cooled to4∘C over 12 h to extract the anthocyanins The solution wasthen centrifuged and the supernatant was frozen and furtherlyophilized Prior to the in vitro and ex vivo experiments thelyophilized material was dissolved in ultrapure water (Milli-Q) at the desired concentrations or doses

23 Total Phenolic Compounds (TPC) The total phenoliccompounds (TPC) in the PT were determined according tothe method described by Singleton and Rossi [14] whichis based on the reduction of the phosphowolframate phos-phomolybdate complex by phenolics to a blue product thatis measured at 750 nm The results are expressed as gallicacid equivalents (mg gallic acid equivalents100 g fresh fruit)

and the values are presented as the means of triplicate analy-sis

24 Total Monomeric Anthocyanins (TMA) The total mon-omeric anthocyanin (TMA) content was determined usingthe pH differential method [15] The anthocyanin con-tent was calculated using the molar absorptivity (120576) andmolecular weights (MW) of cyanidin 3-O-glucoside (120576 =26900 Lmolsdotcm MW = 4492 gmol) The results are ex-pressed as mg of cyanidin 3-O-glucoside equivalents100 gfresh fruit

25 Identification and Quantification of Anthocyanins in PT

251 Chemical Apparatus and General Procedures Allchemicals were of analytical grade Acetonitrile and formicacid were purchased from Merck (Darmstadt Germany)Cyanidin chloride malvidin chloride cyanidin 3-O-gluco-side chloride malvidin 3-O-glucoside chloride and delphini-din 3-O-glucoside chloride were acquired from ChromaDexHigh performance liquid chromatography (HPLC-DAD)wasperformed with a Shimadzu Prominence Auto Sampler (SIL-20A) HPLC system (Shimadzu Kyoto Japan) equipped withShimadzu LC-20AT reciprocating pumps connected to aDGU 20A5 degasser with a CBM 20A integrator SPD-M20Adiode array detector and LC solution 122 SP1 software

252 Quantification of Compounds by HPLC-DAD Reversephase chromatographic analyses were carried out undergradient conditions using a C18 column (46mm times 150mm)packed with 5 120583m diameter particles the mobile phase waswater containing 1 formic acid (A) and acetonitrile (B)and the composition gradient was 13 of B until 10minand changed to obtain 20 30 50 60 70 20 and10 B at 20 30 40 50 60 70 and 80min respectively[16] The P trunciflora aqueous extract and mobile phasewere filtered through a 045 120583m membrane filter (Millipore)and then degassed in an ultrasonic bath prior to use theaqueous extractwas analyzed at a concentration of 20mgmLThe flow rate was 05mLmin the injection volume was20120583L and the wavelength was 520 nm Stock solutions ofstandards references were prepared in the HPLC mobilephase at a concentration range of 0030 to 0250mgmLThe chromatography peaks were confirmed by comparingtheir retention times with those of the reference standardsand DAD spectra (300 to 700 nm) All chromatographyoperations were carried out at ambient temperature and intriplicate [17]

26 Antioxidant Activity In Vitro

261 Determination of Ferric Reducing Antioxidant Power(FRAP) The FRAP assay was based on Benzie and Strain[18] by measuring the absorbance of the complex formedbetween Fe2+ and Ferric-246-tripyridyl-s-triazine (TPTZ)at 593 nm after incubation (37∘C15min) with PT (10ndash160 120583gmL) The increase in the absorbance was comparedto that induced by ascorbic acid (standard) and the results

Evidence-Based Complementary and Alternative Medicine 3

are expressed as the means of the absorbance of triplicateexperiments (119899 = 3)

262 11-Diphenyl-2-2-picrylhydrazyl Radical ScavengingAssay The antiradical powers of the different concentrationsof PT (10ndash160 120583gmL) and standard were determined bymeasuring the decrease in the DPPH absorbance after 24 hin the dark compared to a blank [19] The same procedurewas followed for the ascorbic acid standardThis analysis wascarried out in triplicate (119899 = 3) and the results are expressedas the means of inhibition of the DPPH radical whichwas calculated as follows inhibition = [(Abs control ndashAbs sample)Abs control] times 100 The concentration of PTthat could scavenge 50 of the DPPH radical (IC

50

) wascalculated via a nonlinear regression analysis using theGraphPad Prism Program version 60

263 Protection against Lipid Peroxidation A low-speedsupernatant (20min at 2000timesg) of brain homogenates(50mMTris-HCl pH 75 1 9 wv) was preincubated at 37∘Cfor 1 h in the presence or absence of 50120583MFeCl

2

1 mMH2

O2

PT (10ndash160 120583gmL) and Tris-HCl 50mM Subsequently theamount of thiobarbituric acid-reactive substances (TBARS)was determined [20]The inhibitory concentration 50 (IC

50

)which represents the concentration of PT that inhibits 50 oflipid peroxidation was determined via a nonlinear regressionanalysis using the GraphPad Prism Program version 60

27 In Vivo Studies

271 Animals 80 two-month-old male Swiss mice (30ndash40 g)were obtained from the Bioterism Center of UnochapecoSeven mice were housed per cage (30 times 19 times 13 cm) andmaintained in our own animal facility under controlledenvironmental conditions (22 plusmn 1∘C 12 hr lightdark cyclefree access to food (Nuvilab CR1) and water) All procedureswere carried out in accordance with institutional policies onthe handling of experimental animals (approved by the ethicscommittee process 0012012)

272 Drugs Fluoxetine was used as commercial Daforin(Laboratorio EMS SP Brazil) All drugs were dissolved insaline (NaCl 09) The drugs and saline were administeredintraperitoneally (ip) or orally (po) at a constant volume of01mL10 g body weight

273 Tail Suspension Test (TST) The TST was used asdescribed by Steru et al [21] Mice (119899 = 7ndash10) were orallytreated with vehicle (09 saline wv) or PT (200 400 or800mgkg) An additional group was treated with fluoxetine(32mgkg ip) None of the selected doses modified locomo-tion in the open field test (data not shown) The mice weresubmitted to the TST for 30 or 60min (for ip and po treatedgroups resp) after treatments After the TST the animalswere euthanized and their brains were removed immediatelyin order to assess the oxidative stress parameter ex vivo Theex vivo analyses were performed in the PT 800mgkg group

because the behavioral effects of this dose were comparableto those of fluoxetine

28 Antioxidant Activity Ex Vivo

281 Antioxidant Enzymes The antioxidant activity wasassessed ex vivo using homogenized mouse brains in 7volumes of 50mM Tris buffer (pH 74) The homogenate wascentrifuged at 3000timesg and 4∘C for 10min to yield a low-speedsupernatant for which all parameters were evaluated TheSOD CAT and GPx activity were determined according toMisra and Fridovich [22] Aebi [23] and Paglia and Valentine[24] respectively The TrxR activity was determined using551015840-dithiobis (2-nitrobenzoic acid) (DTNB) and NADPH[25] Gold (III) chloride trihydrate (500 nM) was used toinhibit the thioredoxin reductase activity [26] and determinethe nonthioredoxin reductase DTNB reduction which wassubtracted from the total DTNB reduction in order to obtainthe thioredoxin reductase activity The amount of reducedDTNBwas calculated using an absorption coefficient of 136times103molcm

282 Nonprotein Thiol Groups The low-speed supernatantfraction was mixed with 10 trichloroacetic acid (1 1 vv)followed by the centrifugation and neutralization of thesupernatant (to pH 75) with 1M Tris The nonprotein thiolgroups were immediately determined using a standard curveof cysteine [27]

283 Protein Quantification The protein content was mea-sured using bovine serum albumin as a standard [28]

284 Lipid Peroxidation After the addition of 72mMbutylated hydroxytoluene to prevent further oxidation thesupernatant was used to determine the amount of reactivethiobarbituric acid [20] The samples were extracted with n-butanol and the reaction product was determined at 535 nmusing a standard curve of 1133-tetraethoxypropane

29 Statistical Analysis The results (cumulative counts forspontaneous locomotion time in seconds for immobility andantioxidant activity in vitro and ex vivo) are expressed as themean plusmn SEM Comparisons between the groups were madeby one-way ANOVA followed by Tukeyrsquos post hoc test Thedifferences between the data were analyzed using Studentrsquos119905-test to assess the antioxidant activity in vitro (differentconcentrations of PT times different concentrations of ascorbicacid) Results with 119901 lt 005 were considered significant Theregression analyses were made using Statistica 70 softwaresystem (Statsoft Inc 2001)

3 Results

The content of antioxidant compounds such as total phe-nolics and anthocyanins was quantified The total phenoliccompounds in the aqueous extract of P trunciflora were120167 plusmn 3329mg GAE100 g while the anthocyanins


00 10

1

20

(min)

300

200

100

0

(mAU

)520nm 4nm (100)

(a)

00 10 20

2

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(b)

00 10

3

20

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(c)

00 10

4

20

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(d)

00 10 20

5

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(e)

00 10 20

(min)

750

500

250

0

(mAU

)

520nm 4nm (100)

1

2

3

4

5

(f)

Figure 1 Representative high performance liquid chromatography profiles of the standards cyanidin chloride (a) malvidin chloride (b)delphinidin 3-O-glucoside chloride (c) cyaniding 3-O-glucoside chloride (d) malvidin 3-O-glucoside chloride (e) and PT (f)

content was 17533 plusmn 1180mg cyanidin 3-O-glucoside equiv-alents100 g

The HPLC fingerprinting of the P trunciflora aqueousextract (Figure 1) revealed the presence of anthocyaninsWe identified cyanidin (Rt = 641min peak 1) malvidin(Rt = 973min peak 2) delphinidin 3-O-glucoside (Rt =1194min peak 3) cyanidin 3-O-glucoside (Rt = 1508 peak4) and malvidin 3-O-glucoside (Rt = 2057min peak 5)

The composition of anthocyanins (mgg) in the P truncifloraaqueous extract was cyanidin (162 plusmn 001) malvidin (35 plusmn002) delphinidin 3-O-glycoside (241 plusmn 003) cyanidin 3-O-glycoside (276 plusmn 003) and malvidin 3-O-glycoside (171 plusmn002)

A one-way ANOVA revealed that PT showed signifi-cant reducing power beyond a concentration of 10120583gmL(Figure 2(a)) However the ferric reducing power of ascorbic


0

1

2

3

4

Ascorbic acidPT

Abso

rban

ce

0 10 20 40 80 160

Concentration (120583gmL)

lowast120576

lowastlowast

lowastsect

lowast120595

(a)

0

20

40

60

80

100

Ascorbic acid

PT

DPP

H in

hibi

tion

()

10 20 40 80 160


lowast120574

lowastlowast

lowastlowast

(b)

Figure 2 Ferric reducing antioxidant power (FRAP (a)) andDPPHradical scavenger activity (b) of PTThe results are expressed as the mean plusmnSEM 119899 = 3 (a) lowastDifferent from ascorbic acid solution at the same concentration lowast119901 lt 005 Studentrsquos 119905-test Different symbols representdifferent results within the PT group (119901 lt 001 ANOVATukey) (b) lowastDifferent from DPPH radical scavenger activity of ascorbic acidsolution at the same concentration lowast119901 lt 0001 Studentrsquos 119905-test Different symbols represent different results within the PT group (119901 lt 005ANOVATukey)

acid was higher than that shown by PT in all concentrationsevaluated as evident from Studentrsquos 119905 test Figure 2(b) showsthe DPPH radical scavenging antioxidant activity Althoughthe DPPH radical scavenging ability of PT was lower thanthat of the ascorbic acid solution it was remarkable at allevaluated concentrations The calculated IC

50

value for PTwas 422 120583gmL compared to a value of 004120583gmL forascorbic acid At 10 and 20 120583gmL PT inhibited between11 and 14 of DPPH at 40 120583gmL the inhibition increasedto 50 and exceeded 80 at concentrations of 80 and160 120583gmL

The extract inhibited the lipid peroxidation in ahomogenate of mouse brain at all concentrations (Figure 3)At 10 120583gmL PT inhibited approximately 20 of lipidperoxidation and the inhibition increased to 40 at 20 and40 120583gmL and reached 60 at concentrations of 80 and160 120583gmLThe calculated IC

50

value for PT was 704120583gmLThe results in Figure 4 show the effects of PT (200 400

and 800mgkg po) and fluoxetine (32mgkg ip) duringthe tail suspension test in mice PT significantly reducedthe immobility time in the TST (400 and 800mgkg po119865440

= 48 119901 lt 00001) Fluoxetine significantly reduced theimmobility time in the TST The PT (800mgkg po) wascompared with fluoxetine The spontaneous locomotion ofgroups treated with PT did not differ from the controls (datanot shown)

Figure 5 presents the effect of PT (800mgkg po) andfluoxetine (32mgkg ip) administration on the antioxidantenzyme activities in the homogenate of mouse brains PTand fluoxetine did not result in significant changes in theSOD (Figure 5(a)) GPx (Figure 5(b)) and TrxR (Figure 5(c))

0

20

40

60

80

Inhi

bitio

n of

lipi

d pe

roxi

datio

n (

)

0 10 20 40 80 160


lowast

amp amp

Figure 3 Inhibition of Fenton reaction-induced lipid peroxidationof PT The results are expressed as the means plusmn SEM 119899 =5 lowastDifferent symbols represent different results within the PT group(119901 lt 005 ANOVATukey)

activities compared to the controls fluoxetine significantlyincreased the CAT (Figure 5(d)) activity compared to thecontrols

Figure 6 shows the effect of PT (800mgkg po) and flu-oxetine (32mgkg ip) on the lipid peroxidation and level ofnonprotein thiol groups (NPSH) in the homogenate ofmousebrains Both the PT extract and the fluoxetine attenuated lipidperoxidation (Figure 6(a)) The levels of nonprotein thiol inthe PT extract and fluoxetine (Figure 6(b)) groups did notdiffer from that of the control


0

50

100

150

200

250

Imm

obili

ty ti

me (

s)

lowast

lowastlowast

FLU 32 PT 200 PT 400 PT 800C

Figure 4 Effects of PT (200 400 and 800mgkg po) and fluoxe-tine (32mgkg ip) in the TST Each column represents the mean plusmnSEM 119899 = 7ndash10 lowast119901 lt 00001 times saline ANOVA Tukey

4 Discussion

Depression has been associated with lowered concentra-tions of several endogenous antioxidant compounds such ascoenzyme Q10 vitamins C and E or antioxidant enzymessuch as GPx [29] In addition ROS and RNS have beenshown to modulate neurotransmitter systems involved in theneurobiology of depression [30] In this context this studyintended to evaluate the antidepressant-like effect of a Ptrunciflora (PT) aqueous extract using the TST Moreoverconsidering that jaboticaba species are rich in flavonoids andrelated polyphenols [31] the antioxidant effects of PT wereevaluated by in vitro and ex vivo assays

Our results showed for the first time that oral PT (400and 800mgkg) had antidepressant-like activity in the TSTThis effect was dose related (119903 = minus084 119901 lt 0001Pearson correlation analysis) Furthermore the effect of PT(800mgkg) was comparable to that of the antidepressant flu-oxetine (32mgkg) a selective serotonin reuptake inhibitorTo avoid false positives in the TST our results showed thatPT treatment did not alter locomotor activity in the open fieldtest (Figure 1S in Supplementary Material available online athttpdxdoiorg1011552015601503)

Although the mechanisms of the antidepressant-likeactivity of PT remain unclear the bioactive compoundscurrently identified as well as their antioxidant propertiesmay be involved in this effect Flavonoids such as antho-cyanins stand out among the major classes of phenoliccompounds of plants [32]The cyanidin-3-O-glycoside (peak4) was the dominant anthocyanin present in our extractOther anthocyanins such as delphinidin 3-O-glucoside andcyanidin- 3-O-glucoside were also detected Importantly theantioxidant effects of these compounds have been describedin the literature [33] Data of linear regression revealed asubstantial contribution of TPC and TMA for the reducingantioxidant power of PT assessed by FRAP method (119877 =096 and 119877 = 097 119901 = 000001 resp see Figures 2S-3Ssupplementary data) TPC and TMA also contributed to inhibition of DPPH and inhibition of lipid peroxidation(119877 = 083 and 119877 = 093 resp) as demonstrated

by nonlinear regression (see Figures 4Sndash7S supplementarydata)

The FRAP assay measures the ability of an antioxidantsubstance to donate one electron [18] Because the antioxi-dant activity of a substance correlates with its reducing prop-erties the reduction of the 246-tripyridyl-s-triazine-Fe(III)complex due to PT indicates the presence of compounds thatcan donate electrons such as phenolic compounds Accord-ingly the antioxidant properties of the Syzygium cumini fruitskin may in part be due to the antioxidant vitamins tanninsphenolics and anthocyanin compounds present in the fruit[34]

The reducing power of PT was corroborated by theDPPH radical scavenging assay which also evaluates theability of antioxidants to transfer a single electron Thisaffirmation is based on the fact that both the reducing andthe scavenging DPPH abilities of the extract were observedin the entire evaluated concentration range (10ndash160120583gmL)Therefore these results strongly suggest that the DPPHradical scavenging capacity of PT is related to its reducingproperties as evidenced in the FRAP assay Conversely PTcould not remove H

2

O2

or O2∙minus nor avoid the H

2

O2

-inducedoxidation of GSH (data not shown)

Lipid peroxidation is an index of oxidative stress andmay result in damage to components of the cell membranewhich may lead to calcium influx and cell death Lipidperoxidation is associated with several diseases includingneurodegenerative disorders [35] and antioxidants may pro-tect against lipid peroxidation by scavenging the free radicals[36] The in vitro results of the current study showed thatPT inhibited the lipid peroxidation at all concentrationsThis protective effect suggests that other possiblemechanismsof action of the antioxidant activity are associated withthe ability of this extract to scavenge the hydroxyl (∙OH)radical Interestingly PT (800mgkg) also attenuated lipidperoxidation when administered to mice This protectionwas similar to that observed for fluoxetine (32mgkg) andoccurred at a dose that showed an antidepressant-like effectFluoxetine decreased lipid peroxidation probably due to theincreased CAT activity which removes H

2

O2

to reduce itsavailability for the formation of the ∙OH radical Similarlyfluoxetine exerted a restorative action on the oxidativeeffects in the peripheral defense cells of animals submittedto the restraint stress model which was also associatedwith enhanced endogenous antioxidant defenses (CAT andSOD) and the restoration of GSH levels [37] Oxidativedamage to lipids and decreased antioxidant enzyme activ-ity have been reported in patients with major depressivedisorder [38] and preclinical studies have suggested thatantioxidants may have antidepressant properties [39] Takinginto account these findings the inhibition of lipid perox-idation by PT as well as the ability of PT to scavengefree radicals strongly suggests a link between the antiox-idant activity and the antidepressant-like effects observedhere

The present study showed the in vitro and ex vivo antiox-idant and antidepressant-like effects of PT in mice Theseantioxidant propertiesmight be related to the antidepressant-like activity of Plinia trunciflora


0

10

20

30

40SO

D (U

mg

prot

ein)

FLU PT 800C

(a)

000

002

004

006

008

010

GPx

(nm

ol N

AD

PHm

inm

g pr

otei

n)

FLU PT 800C

(b)

0

10

20

30

40

TrxR

(nm

ol D

TNB

min

mg

prot

ein)

FLU PT 800C

(c)

0

2

4

6

CAT

(kg

pro

tein

)lowast

FLU PT 800C

(d)

Figure 5 Effects of PT (800mgkg po) and fluoxetine (32mgkg ip) treatment on antioxidant enzyme activities SOD (a) GPx (b) TrxR(c) and CAT (d) in homogenate of brain mice The results are expressed as the means plusmn SEM 119899 = 6 lowast119901 lt 005 times saline ANOVATukey

lowast

lowast

00

01

02

03

TBA

RS (n

mol

MD

Am

g pr

otei

n)

FLU PT 800C

(a)

0

5

10

15

20

NPS

H (n

mol

NPS

Hm

g pr

otei

n)

FLU PT 800C

(b)

Figure 6 Effects of PT (800mgkg po) and fluoxetine (32mgkg ip) on lipid peroxidation (a) and nonprotein thiol groups (NPSH) level(b) The results are expressed as the means plusmn SEM 119899 = 6 lowast119901 lt 005 times saline ANOVATukey


Conflict of Interests

The authors have declared that no competing interests exist

References

[1] A J Ferrari F J Charlson R E Norman et al ldquoBurden ofdepressive disorders by country sex age and year findingsfrom the global burden of disease study 2010rdquo PLoS Medicinevol 10 no 11 Article ID e1001547 2013

[2] A J Smith I Sketris C Cooke D Gardner S Kisely and SE Tett ldquoA comparison of antidepressant use in Nova ScotiaCanada andAustraliardquo Pharmacoepidemiology andDrug Safetyvol 17 no 7 pp 697ndash706 2008

[3] J J Schildkraut E K Gordon and J Durell ldquoCatecholaminemetabolism in affective disorders I Normetanephrine andVMA excretion in depressed patients treated with imipraminerdquoJournal of Psychiatric Research vol 3 no 4 pp 213ndash228 1965

[4] T M Michel S Frangou D Thiemeyer et al ldquoEvidence foroxidative stress in the frontal cortex in patients with recurrentdepressive disordermdasha postmortem studyrdquo Psychiatry Researchvol 151 no 1-2 pp 145ndash150 2007

[5] J Nordberg and E S J Arner ldquoReactive oxygen speciesantioxidants and the mammalian thioredoxin systemrdquo FreeRadical Biology andMedicine vol 31 no 11 pp 1287ndash1312 2001

[6] B Halliwell ldquoRole of free radicals in the neurodegenerativediseases therapeutic implications for antioxidant treatmentrdquoDrugs and Aging vol 18 no 9 pp 685ndash716 2001

[7] S V Avery ldquoMolecular targets of oxidative stressrdquo BiochemicalJournal vol 434 no 2 pp 201ndash210 2011

[8] P Mecocci and M C Polidori ldquoAntioxidant clinical trials inmild cognitive impairment and Alzheimerrsquos diseaserdquo Biochim-ica et Biophysica Acta vol 1822 no 5 pp 631ndash638 2012

[9] M Valko D Leibfritz J Moncol M T D Cronin M Mazurand J Telser ldquoFree radicals and antioxidants in normal physi-ological functions and human diseaserdquo International Journal ofBiochemistry and Cell Biology vol 39 no 1 pp 44ndash84 2007

[10] C Vollert M Zagaar I Hovatta et al ldquoExercise prevents sleepdeprivation-associated anxiety-like behavior in rats poten-tial role of oxidative stress mechanismsrdquo Behavioural BrainResearch vol 224 no 2 pp 233ndash240 2011

[11] D J Mabberley The Plant Book Cambridge University PressCambridge UK 2nd edition 1997

[12] L C Stasi and C A Hiruma-Lima ldquoMyrtales medicinaisrdquo inPlantas Medicinais na Amazonia e naMata Atlantica L C Stasiand C A Hiruma-Lima Eds pp 321ndash330 Editora UNESP SaoPaulo Brazil 2nd edition 2002

[13] EM Kuskoski A G Asuero AM Troncoso J Mancini-Filhoand R Fett ldquoAplicacion de diversos metodos quımicos paradeterminar actividad antioxidante en pulpa de frutosrdquo Cienciae Tecnologia de Alimentos vol 25 no 4 pp 726ndash732 2005

[14] V L Singleton and J A J Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungstic acid reagentsrdquoAmeri-can Journal of Enology and Viticulture vol 16 no 3 pp 144ndash1581965

[15] J Lee R W Durst and R E Wrolstad ldquoDetermination oftotal monomeric anthocyanin pigment content of fruit juicesbeverages natural colorants and wines by the pH differentialmethod collaborative studyrdquo Journal of AOAC Internationalvol 88 no 5 pp 1269ndash1278 2005

[16] J P Kamdem E O OlalekanW Hassan et al ldquoTrichilia catigua(Catuaba) bark extract exerts neuroprotection against oxidativestress induced by different neurotoxic agents in rat hippocampalslicesrdquo Industrial Crops and Products vol 50 pp 625ndash632 2013

[17] A A Boligon T F Kubica D N Mario et al ldquoAntimicrobialand antiviral activity-guided fractionation from Scutia buxifoliaReissek extractsrdquoActa Physiologiae Plantarum vol 35 no 7 pp2229ndash2239 2013

[18] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996

[19] W Brand-Williams M E Cuvelier and C Berset ldquoUse of a freeradical method to evaluate antioxidant activityrdquo LWTmdashFoodScience and Technology vol 28 no 1 pp 25ndash30 1995

[20] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[21] L Steru R Chermat B Thierry and P Simon ldquoThe tailsuspension test a new method for screening antidepressants inmicerdquo Psychopharmacology vol 85 no 3 pp 367ndash370 1985

[22] H PMisra and I Fridovich ldquoThe role of superoxide anion in theautoxidation of epinephrine and a simple assay for superoxidedismutaserdquo Journal of Biological Chemistry vol 247 no 10 pp3170ndash3175 1972

[23] H Aebi ldquoCatalase in vitrordquoMethods in Enzymology vol 105 pp121ndash126 1984

[24] D E Paglia and W N Valentine ldquoStudies on the quantitativeand qualitative characterization of erythrocyte glutathione per-oxidaserdquo The Journal of Laboratory and Clinical Medicine vol70 no 1 pp 158ndash169 1967

[25] A Holmgren andM Bjornstedt ldquoThioredoxin and thioredoxinreductaserdquoMethods in Enzymology vol 252 pp 199ndash208 1995

[26] Y OmataM FolanM Shaw et al ldquoSublethal concentrations ofdiverse gold compounds inhibit mammalian cytosolic thiore-doxin reductase (TrxR1)rdquo Toxicology In Vitro vol 20 no 6 pp882ndash890 2006

[27] G L Ellman ldquoTissue sulfhydryl groupsrdquo Archives of Biochem-istry and Biophysics vol 82 no 1 pp 70ndash77 1959

[28] O H Lowry N J Rosebrough A L Farr and R J RandallldquoProtein measurement with the Folin phenol reagentrdquo TheJournal of Biological Chemistry vol 193 no 1 pp 265ndash275 1951

[29] M Maes P Galecki Y S Chang and M Berk ldquoA reviewon the oxidative and nitrosative stress (OampNS) pathwaysin major depression and their possible contribution to the(neuro)degenerative processes in that illnessrdquo Progress inNeuro-Psychopharmacology and Biological Psychiatry vol 35no 3 pp 676ndash692 2011

[30] V O Kotan E Sarandol E Kirhan G Ozkaya and S KirlildquoEffects of long-term antidepressant treatment on oxidativestatus in major depressive disorder a 24-week follow-up studyrdquoProgress in Neuro-Psychopharmacology and Biological Psychia-try vol 35 no 5 pp 1284ndash1290 2011

[31] K A Reynertson A M Wallace S Adachi et al ldquoBioac-tive depsides and anthocyanins from jaboticaba (Myrciariacauliflora)rdquo Journal of Natural Products vol 69 no 8 pp 1228ndash1230 2006

[32] S Aparecida de Assis J C R Vellosa I L Brunetti et alldquoAntioxidant activity ascorbic acid and total phenol of exoticfruits occurring in Brazilrdquo International Journal of Food Sciencesand Nutrition vol 60 no 5 pp 439ndash448 2009


[33] A Castaneda-Ovando M D L Pacheco-Hernandez M EPaez-Hernandez J A Rodrıguez and C A Galan-VidalldquoChemical studies of anthocyanins a reviewrdquo Food Chemistryvol 113 no 4 pp 859ndash871 2009

[34] A Banerjee N Dasgupta and B De ldquoIn vitro study ofantioxidant activity of Syzygium cumini fruitrdquo Food Chemistryvol 90 no 4 pp 727ndash733 2005

[35] B Uttara A V Singh P Zamboni and R T MahajanldquoOxidative stress and neurodegenerative diseases a review ofupstream and downstream antioxidant therapeutic optionsrdquoCurrent Neuropharmacology vol 7 no 1 pp 65ndash74 2009

[36] M Z Gul L M Bhakshu F Ahmad A K Kondapi IA Qureshi and I A Ghazi ldquoEvaluation of Abelmoschusmoschatus extracts for antioxidant free radical scavengingantimicrobial and antiproliferative activities using in vitroassaysrdquo BMC Complementary and Alternative Medicine vol 11article 64 2011

[37] S Novıo M J Nunez G Amigo and M Freire-GaraballdquoEffects of fluoxetine on the oxidative status of peripheralblood leucocytes of restraint-stressed micerdquo Basic and ClinicalPharmacology and Toxicology vol 109 no 5 pp 365ndash371 2011

[38] A Sarandol E Sarandol S S Eker S Erdinc E Vatansever andS Kirli ldquoMajor depressive disorder is accompanied with oxida-tive stress short-term antidepressant treatment does not alteroxidativendashantioxidative systemsrdquo Human Psychopharmacologyvol 22 no 2 pp 67ndash73 2007

[39] A Zafir A Ara and N Banu ldquoIn vivo antioxidant status aputative target of antidepressant actionrdquo Progress in Neuro-Psychopharmacology and Biological Psychiatry vol 33 no 2 pp220ndash228 2009

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


as catalase (CAT) superoxide dismutase (SOD) glutathioneperoxidase (GPx) and thioredoxin reductase (TrxR) [5]The overproduction of ROS and oxidative stress have beenimplicated in the pathophysiological processes related tovarious diseases including Alzheimerrsquos Parkinsonrsquos anx-iety and depression [8ndash10] In this sense plants emergeas potential alternatives for the treatment of oxidativestress-related diseases considering that they are importantsources of carotenoids flavonoids vitamins and polyphe-nols

TheMyrtaceae family consists of 4620 species distributedin 140 genera whose occurrence has been described insubtropical and tropical regions of the world mainly Cen-tral and South America and Australia [11] The jaboticabatree Plinia trunciflora (O Berg) Kausel a synonym ofMyrciaria trunciflora O Berg Eugenia cauliflora O Bergand Myrciaria peruviana (Poir) Mattos is popularly namedldquojabuticabeirardquo in Brazil (source httpwwwtropicosorg)In folk medicine species of Plinia have been used to treatvarious diseases such as diabetes and chronic inflammationof the tonsils [12] However studies that evaluate the effects ofthis species on the central nervous system (CNS) are scarce inthe literature Therefore the aim of this study was to evaluatethe antidepressant-like effect of P trunciflora aqueous extractin the tail suspension test The identification of bioactivecompounds and the in vitro and ex vivo antioxidant effectsof P trunciflora were investigated in order to establish if theantidepressant-like effect of this plant could be related tothese properties

2 Materials and Methods

21 Plant Material The whole fruits of Plinia trunciflorawere collected in Alpestre (RS Brazil) (27∘101015840568210158401015840S and53∘71015840195510158401015840O) in September and taxonomically identified byMarcos Eduardo Guerra Sobral (botanical) where a voucherhas been deposited in the university herbarium (number3302)

22 Preparation of Aqueous Extract of P trunciflora (PT)The aqueous extracts of whole fruits were prepared basedon the methodology described by Kuskoski et al [13] Thewhole fruit (100 g) of P trunciflora was mixed with 200mLof distilled water and acidified with concentrated HCl untilpH 15 After trituration for 1 min the solution was cooled to4∘C over 12 h to extract the anthocyanins The solution wasthen centrifuged and the supernatant was frozen and furtherlyophilized Prior to the in vitro and ex vivo experiments thelyophilized material was dissolved in ultrapure water (Milli-Q) at the desired concentrations or doses

23 Total Phenolic Compounds (TPC) The total phenoliccompounds (TPC) in the PT were determined according tothe method described by Singleton and Rossi [14] whichis based on the reduction of the phosphowolframate phos-phomolybdate complex by phenolics to a blue product thatis measured at 750 nm The results are expressed as gallicacid equivalents (mg gallic acid equivalents100 g fresh fruit)

and the values are presented as the means of triplicate analy-sis

24 Total Monomeric Anthocyanins (TMA) The total mon-omeric anthocyanin (TMA) content was determined usingthe pH differential method [15] The anthocyanin con-tent was calculated using the molar absorptivity (120576) andmolecular weights (MW) of cyanidin 3-O-glucoside (120576 =26900 Lmolsdotcm MW = 4492 gmol) The results are ex-pressed as mg of cyanidin 3-O-glucoside equivalents100 gfresh fruit

25 Identification and Quantification of Anthocyanins in PT

251 Chemical Apparatus and General Procedures Allchemicals were of analytical grade Acetonitrile and formicacid were purchased from Merck (Darmstadt Germany)Cyanidin chloride malvidin chloride cyanidin 3-O-gluco-side chloride malvidin 3-O-glucoside chloride and delphini-din 3-O-glucoside chloride were acquired from ChromaDexHigh performance liquid chromatography (HPLC-DAD)wasperformed with a Shimadzu Prominence Auto Sampler (SIL-20A) HPLC system (Shimadzu Kyoto Japan) equipped withShimadzu LC-20AT reciprocating pumps connected to aDGU 20A5 degasser with a CBM 20A integrator SPD-M20Adiode array detector and LC solution 122 SP1 software

252 Quantification of Compounds by HPLC-DAD Reversephase chromatographic analyses were carried out undergradient conditions using a C18 column (46mm times 150mm)packed with 5 120583m diameter particles the mobile phase waswater containing 1 formic acid (A) and acetonitrile (B)and the composition gradient was 13 of B until 10minand changed to obtain 20 30 50 60 70 20 and10 B at 20 30 40 50 60 70 and 80min respectively[16] The P trunciflora aqueous extract and mobile phasewere filtered through a 045 120583m membrane filter (Millipore)and then degassed in an ultrasonic bath prior to use theaqueous extractwas analyzed at a concentration of 20mgmLThe flow rate was 05mLmin the injection volume was20120583L and the wavelength was 520 nm Stock solutions ofstandards references were prepared in the HPLC mobilephase at a concentration range of 0030 to 0250mgmLThe chromatography peaks were confirmed by comparingtheir retention times with those of the reference standardsand DAD spectra (300 to 700 nm) All chromatographyoperations were carried out at ambient temperature and intriplicate [17]

26 Antioxidant Activity In Vitro

261 Determination of Ferric Reducing Antioxidant Power(FRAP) The FRAP assay was based on Benzie and Strain[18] by measuring the absorbance of the complex formedbetween Fe2+ and Ferric-246-tripyridyl-s-triazine (TPTZ)at 593 nm after incubation (37∘C15min) with PT (10ndash160 120583gmL) The increase in the absorbance was comparedto that induced by ascorbic acid (standard) and the results




50



2

1 mMH2

O2


50


27 In Vivo Studies











3 Results



00 10

1

20

(min)

300

200

100

0

(mAU

)520nm 4nm (100)

(a)

00 10 20

2

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(b)

00 10

3

20

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(c)

00 10

4

20

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(d)

00 10 20

5

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(e)

00 10 20

(min)

750

500

250

0

(mAU

)

520nm 4nm (100)

1

2

3

4

5

(f)







0

1

2

3

4

Ascorbic acidPT

Abso

rban

ce

0 10 20 40 80 160


lowast120576

lowastlowast

lowastsect

lowast120595

(a)

0

20

40

60

80

100

Ascorbic acid

PT

DPP

H in

hibi

tion

()

10 20 40 80 160


lowast120574

lowastlowast

lowastlowast

(b)



50



50





0

20

40

60

80

Inhi

bitio

n of

lipi

d pe

roxi

datio

n (

)

0 10 20 40 80 160


lowast

amp amp





0

50

100

150

200

250

Imm

obili

ty ti

me (

s)

lowast

lowastlowast

FLU 32 PT 200 PT 400 PT 800C


4 Discussion







2

O2


2

O2



2

O2




0

10

20

30

40SO

D (U

mg

prot

ein)

FLU PT 800C

(a)

000

002

004

006

008

010

GPx

(nm

ol N

AD

PHm

inm

g pr

otei

n)

FLU PT 800C

(b)

0

10

20

30

40

TrxR

(nm

ol D

TNB

min

mg

prot

ein)

FLU PT 800C

(c)

0

2

4

6

CAT

(kg

pro

tein

)lowast

FLU PT 800C

(d)


lowast

lowast

00

01

02

03

TBA

RS (n

mol

MD

Am

g pr

otei

n)

FLU PT 800C

(a)

0

5

10

15

20

NPS

H (n

mol

NPS

Hm

g pr

otei

n)

FLU PT 800C

(b)





References














































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















50



2

1 mMH2

O2


50


27 In Vivo Studies











3 Results



00 10

1

20

(min)

300

200

100

0

(mAU

)520nm 4nm (100)

(a)

00 10 20

2

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(b)

00 10

3

20

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(c)

00 10

4

20

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(d)

00 10 20

5

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(e)

00 10 20

(min)

750

500

250

0

(mAU

)

520nm 4nm (100)

1

2

3

4

5

(f)







0

1

2

3

4

Ascorbic acidPT

Abso

rban

ce

0 10 20 40 80 160


lowast120576

lowastlowast

lowastsect

lowast120595

(a)

0

20

40

60

80

100

Ascorbic acid

PT

DPP

H in

hibi

tion

()

10 20 40 80 160


lowast120574

lowastlowast

lowastlowast

(b)



50



50





0

20

40

60

80

Inhi

bitio

n of

lipi

d pe

roxi

datio

n (

)

0 10 20 40 80 160


lowast

amp amp





0

50

100

150

200

250

Imm

obili

ty ti

me (

s)

lowast

lowastlowast

FLU 32 PT 200 PT 400 PT 800C


4 Discussion







2

O2


2

O2



2

O2




0

10

20

30

40SO

D (U

mg

prot

ein)

FLU PT 800C

(a)

000

002

004

006

008

010

GPx

(nm

ol N

AD

PHm

inm

g pr

otei

n)

FLU PT 800C

(b)

0

10

20

30

40

TrxR

(nm

ol D

TNB

min

mg

prot

ein)

FLU PT 800C

(c)

0

2

4

6

CAT

(kg

pro

tein

)lowast

FLU PT 800C

(d)


lowast

lowast

00

01

02

03

TBA

RS (n

mol

MD

Am

g pr

otei

n)

FLU PT 800C

(a)

0

5

10

15

20

NPS

H (n

mol

NPS

Hm

g pr

otei

n)

FLU PT 800C

(b)





References














































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















00 10

1

20

(min)

300

200

100

0

(mAU

)520nm 4nm (100)

(a)

00 10 20

2

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(b)

00 10

3

20

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(c)

00 10

4

20

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(d)

00 10 20

5

(min)

300

200

100

0

(mAU

)

520nm 4nm (100)

(e)

00 10 20

(min)

750

500

250

0

(mAU

)

520nm 4nm (100)

1

2

3

4

5

(f)







0

1

2

3

4

Ascorbic acidPT

Abso

rban

ce

0 10 20 40 80 160


lowast120576

lowastlowast

lowastsect

lowast120595

(a)

0

20

40

60

80

100

Ascorbic acid

PT

DPP

H in

hibi

tion

()

10 20 40 80 160


lowast120574

lowastlowast

lowastlowast

(b)



50



50





0

20

40

60

80

Inhi

bitio

n of

lipi

d pe

roxi

datio

n (

)

0 10 20 40 80 160


lowast

amp amp





0

50

100

150

200

250

Imm

obili

ty ti

me (

s)

lowast

lowastlowast

FLU 32 PT 200 PT 400 PT 800C


4 Discussion







2

O2


2

O2



2

O2




0

10

20

30

40SO

D (U

mg

prot

ein)

FLU PT 800C

(a)

000

002

004

006

008

010

GPx

(nm

ol N

AD

PHm

inm

g pr

otei

n)

FLU PT 800C

(b)

0

10

20

30

40

TrxR

(nm

ol D

TNB

min

mg

prot

ein)

FLU PT 800C

(c)

0

2

4

6

CAT

(kg

pro

tein

)lowast

FLU PT 800C

(d)


lowast

lowast

00

01

02

03

TBA

RS (n

mol

MD

Am

g pr

otei

n)

FLU PT 800C

(a)

0

5

10

15

20

NPS

H (n

mol

NPS

Hm

g pr

otei

n)

FLU PT 800C

(b)





References














































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















0

1

2

3

4

Ascorbic acidPT

Abso

rban

ce

0 10 20 40 80 160


lowast120576

lowastlowast

lowastsect

lowast120595

(a)

0

20

40

60

80

100

Ascorbic acid

PT

DPP

H in

hibi

tion

()

10 20 40 80 160


lowast120574

lowastlowast

lowastlowast

(b)



50



50





0

20

40

60

80

Inhi

bitio

n of

lipi

d pe

roxi

datio

n (

)

0 10 20 40 80 160


lowast

amp amp





0

50

100

150

200

250

Imm

obili

ty ti

me (

s)

lowast

lowastlowast

FLU 32 PT 200 PT 400 PT 800C


4 Discussion







2

O2


2

O2



2

O2




0

10

20

30

40SO

D (U

mg

prot

ein)

FLU PT 800C

(a)

000

002

004

006

008

010

GPx

(nm

ol N

AD

PHm

inm

g pr

otei

n)

FLU PT 800C

(b)

0

10

20

30

40

TrxR

(nm

ol D

TNB

min

mg

prot

ein)

FLU PT 800C

(c)

0

2

4

6

CAT

(kg

pro

tein

)lowast

FLU PT 800C

(d)


lowast

lowast

00

01

02

03

TBA

RS (n

mol

MD

Am

g pr

otei

n)

FLU PT 800C

(a)

0

5

10

15

20

NPS

H (n

mol

NPS

Hm

g pr

otei

n)

FLU PT 800C

(b)





References














































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















0

50

100

150

200

250

Imm

obili

ty ti

me (

s)

lowast

lowastlowast

FLU 32 PT 200 PT 400 PT 800C


4 Discussion







2

O2


2

O2



2

O2




0

10

20

30

40SO

D (U

mg

prot

ein)

FLU PT 800C

(a)

000

002

004

006

008

010

GPx

(nm

ol N

AD

PHm

inm

g pr

otei

n)

FLU PT 800C

(b)

0

10

20

30

40

TrxR

(nm

ol D

TNB

min

mg

prot

ein)

FLU PT 800C

(c)

0

2

4

6

CAT

(kg

pro

tein

)lowast

FLU PT 800C

(d)


lowast

lowast

00

01

02

03

TBA

RS (n

mol

MD

Am

g pr

otei

n)

FLU PT 800C

(a)

0

5

10

15

20

NPS

H (n

mol

NPS

Hm

g pr

otei

n)

FLU PT 800C

(b)





References














































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















0

10

20

30

40SO

D (U

mg

prot

ein)

FLU PT 800C

(a)

000

002

004

006

008

010

GPx

(nm

ol N

AD

PHm

inm

g pr

otei

n)

FLU PT 800C

(b)

0

10

20

30

40

TrxR

(nm

ol D

TNB

min

mg

prot

ein)

FLU PT 800C

(c)

0

2

4

6

CAT

(kg

pro

tein

)lowast

FLU PT 800C

(d)


lowast

lowast

00

01

02

03

TBA

RS (n

mol

MD

Am

g pr

otei

n)

FLU PT 800C

(a)

0

5

10

15

20

NPS

H (n

mol

NPS

Hm

g pr

otei

n)

FLU PT 800C

(b)





References














































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















References














































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





























of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















Antidepressant-Like and Antioxidant Effects of Plinia trunciflora in Mice

Documents

Transcript of Antidepressant-Like and Antioxidant Effects of Plinia trunciflora in Mice