Research Article Acetone Extract from Rhodomyrtustomentosa: A Potent Natural Antioxidant

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1 Evidence-Bsed Complementry nd Alterntive Medicine Volume 212, Article ID , 8 pges doi:1.1155/212/ Reserch Article Acetone Extrct from Rhodomyrtustomentos: A Potent Nturl Antioxidnt Goodl Lvny, 1 Supyng Piywn Vorvuthikunchi, 1, 2 nd Nongporn Hutdilok Towtn 1, 3 1 Nturl Products Reserch Centre, Prince of Songkl University, Ht Yi, Songkhl 9112, Thilnd 2 Deprtment of Microiology, Prince of Songkl University, Ht Yi, Songkhl 9112, Thilnd 3 Deprtment of Biochemistry, Prince of Songkl University, Ht Yi, Songkhl 9112, Thilnd Correspondence should e ddressed to Supyng Piywn Vorvuthikunchi, supyng.v@psu.c.th Received 19 April 212; Accepted 18 June 212 Acdemic Editor: Vssy Bnkov Copyright 212 Goodl Lvny et l. This is n open ccess rticle distriuted under the Cretive Commons Attriution License, which permits unrestricted use, distriution, nd reproduction in ny medium, provided the originl work is properly cited. Rhodomyrtus tomentos (Myrtcee) hs een employed in trditionl Thi medicine to tret colic dirrhoe, dysentery, scesses, hemorrhge, nd gynecopthy. In ddition, it hs een used to formulte skin-whitening, nti-ging nd skin eutifying gents. Ethnomedicl ctivities of this plnt my e due its ntioxidnt property. Hence, the im of this study ws to evlute oth in vitro nd in vivo ntioxidnt ctivities of R. tomentos lef extrct. In vitro ntioxidnt ctivity of the extrct ws ssessed y lipid peroxidtion inhiition cpcity, ferric reducing ntioxidnt power, nd metl chelting ctivity. R. tomentos extrct demonstrted its free rdicl scvenging effects in concentrtion dependent mnner. In vivo ntioxidnt ctivity of the extrct ws conducted in Swiss Alino mice. Levels of thio-rituric cid rective sustnces (TBARS), glutthione (GSH), nd the ctivities of ntioxidnt enzymes including superoxide dismutse (SOD), ctlse (CAT), nd glutthione peroxidse (GPx) in lood, liver, nd kidney were nlyzed using microtitre plte photometer. Administrtion of CCl 4 cused significnt increse in TBARS nd decrese in GSH, SOD, CAT nd GPx levels. In contrst, R. tomentos extrct (.8 g/kg) effectively prevented these ltertions nd mintined the ntioxidnt sttus. The results suggest tht R. tomentos extrct cn serve s potent ntioxidnt. 1. Introduction Oxidtive stress is elieved to e one of the mjor cuses of more thn hundred types of humn diseses s it cn result in severe cellulr dysfunction due to peroxidtion of memrne lipids, protein modifiction, depletion of nicotinmide nucleotides, rises in intrcellulr free clcium ions (C 2+ ), cytoskeletl disruption, nd DNA dmge [1]. Antioxidnts re the molecules tht cn dely or prevent the oxidtion of cellulr sustrtes [2]. Nturl ntioxidnts minly derived from plnts re preferred over synthetic ntioxidnts, s mny of them including propylgllte, citric cid, utylted hydroxynisole, nd utylted hydroxytoluene re suspected to e heptotoxic nd crcinogenic [3]. Rhodomyrtus tomentos (Aiton) Hssk. (Fmily Myrtcee) is n ornmentl, evergreen shru grows up to four meters tll. This plnt species is ntive to southern nd southestern Asi [4]. It hs een often used in trditionl Thi medicine to tret colic dirrhoe [5], dysentery, scesses, hemorrhge, nd gynecopthy [6]. In ddition, it is used to formulte skin-whitening, ntiging, nd skineutifying gents [7]. The extrct from this plnt possess strong inhiitory ctivity ginst grm-positive cteri [8, 9]. A rnge of compounds including cylphloroglucinol, flvonoids, tnnins, nd triterpenes hve een identified from this plnt [1]. Rhodomyrtone, n cylphloroglucinol component from this plnt, hs een recently reported s n effective nti-infective gent ginst Streptococcus pyogenes [11], Stphylococcus ureus, nd iofilm-forming Stphylococci [12]. Although Rhodomyrtus tomentos extrct hs een extensively investigted for its ntimicroil properties [13 16], less or no dt regrding its ntioxidnt properties re ville. It is elieved tht some of the ethnomedicl nd

2 2 Evidence-Bsed Complementry nd Alterntive Medicine reported iologicl ctivities of this plnt my e due its ntioxidnt property. Hence, the present study ws imed to evlute oth in vitro nd in vivo ntioxidnt ctivities of the extrct from Rhodomyrtus tomentos leves. 2. Mteril nd Methods 2.1. Chemicls. Thiorituric cid (TBA), 2,4,6-tripyridyls-trizine (TPTZ), nd ferrozine were purchsed from Sigm-Aldrich Co., USA. Gllic cid, ellgic cid, nd olive oil were otined from Fluk Chemie GmH CH Buchs, Spin. Regents for mesuring Thio-rituric cid recting sustnces (TBARS), totl glutthione (GSH), superoxide dismutse (SOD), ctlse (CAT), nd glutthione peroxidse (GPx) were purchsed from Cymn Chemicl Compny, USA. All other chemicls used were nlyticl grde Plnt Mteril. R. tomentos leves were collected from the Singh Nkorn District, Songkhl Province in the southern prt of Thilnd during Ferury 21. The voucher specimen (A. Hirnrt 1) ws identified y J. Wi nd hs een deposited in the herrium of the Deprtment of Biology, Fculty of Science, Prince of Songkl University, Thilnd Extrct Preprtion. R. tomentos leves were dried in oven t 6 C for 48 h nd grounded in n electric lender. The powder ws extrcted with 95% cetone nd left t room temperture for 7 dys. The extrct ws evported y using rotry evportor (BUCHI Rotvpor R-114, Buchi Lortechnik AG, Flwil, Switzerlnd) nd kept t 4 C In Vitro Studies Lipid Peroxidtion Inhiitory Assy. Lipid peroxidtion induced y Fe 2+ in known concentrtion of liposome ws estimted s TBARS ccording to Ohkw s method [17]. The rection mixture contined liposome from soyen phosphotidylcholine.1 ml (.2 g/l) in Tris-HCl uffer (2 mm, ph 7.), KCl (3 mm), FeSO 4 (NH 4 ) 2 SO 4 (.6 mm), nd vrious concentrtions of R. tomentos extrct in finl volume of.5 ml. After the rection mixture ws incuted t the 37 C for 1 h,.4 ml of the rection mixture ws treted with.2 ml sodium dodecyl sulfte (SDS, 8.1%), 1.5 ml TBA (.8%), nd 1.5 ml cetic cid (2%, ph 3.5). The totl volume ws mde up to 4. ml with distilled wter nd then kept in wter th t 95 to 1 C for 1 h. After cooling, 1. ml of distilled wter nd 5. ml of n-utnol nd pyridine mixture (15 : 1 v/v) were dded to the rection mixture, shken vigorously nd centrifuged t 1912 g for 1 min (SORVALL RC5C PLUS centrifuge, USA). The utnol-pyridine lyer ws removed, nd its sornce t 532 nm ws mesured to quntify TBARS. Inhiition of lipid peroxidtion ws determined y compring the opticl density of smples with tht of stndrd curve of gllic cid Ferric Reducing Antioxidnt Power (FRAP). FRAP ws performed ccording to Erel s method [18] with slight modifictions. Briefly, the working FRAP regent ws prepred y mixing 3 mm cette uffer (ph 3.6), 1 mm TPTZ in 4 mm HCl solution, nd 2 mm FeCl 3.6H 2 O in 1:1:1 rtio, just efore use nd heted to 37 C. A totl of 15 μl working FRAP regent ws dded to ech well in 96-well microtiter plte. A lnk reding ws tken t 593 nm using micro titre plte reder (ELX 88, BioTek Instruments, Inc. USA). A totl of 2 μl of the smple (1 g/l of plnt extrct), nd the stndrds (gllic nd ellgic cids) were then dded to wells contining FRAP regent. The mixture ws then incuted t 37 Cfor 1 min, nd the sornce ws mesured t 593 nm for second time. The chnge in sornce from the initil lnk reding ws then compred with stndrd curve. Stndrds were run t different concentrtions (25, 5, 75, 1, 2, 4, 6, 8, nd 1 μm). A stndrd curve ws then prepred y plotting FRAP vlue of ech stndrd versus its concentrtions. The vlues were expressed s mm of gllic cid nd ellgic cid equivlents per 1 mg extrct Chelting Activity on Ferrous Ions (Fe 2+ ). Fe 2+ chelting ctivity ws mesured y inhiition of the formtion of Fe 2+ -ferrozine complex fter tretment of test mteril with Fe 2+, following Dinis method [19]. Fe 2+ -chelting ility of R. tomentos extrct ws monitored y the sornce of the ferrous ion-ferrozine complex t 562 nm. Briefly, different concentrtions of R. tomentos extrct (2 1μg/mL) in 1%DMSO(.4mL)wereddedtosolutionof.6mM FeCl 2 (.1 ml). The rection ws initited y the ddition of 5 mm ferrozine (.1 ml) dissolved in methnol. Then the mixture ws shken vigorously nd left t room temperture for 1 min. Asornce of the solution ws then mesured spectrophotometriclly (Perkin Elmer Lmd25 UV/VIS spectrometer) t 562 nm. The control contins 1% DMSO, FeCl 2, nd ferrozine. The ferrous ion-chelting effect of R. tomentos extrct ws determined s EDTA equivlent In Vivo Studies Experimentl Animls. Forty Swiss lino mice (Mus musculus) weighing g of the mle sex were otined from the Lortory Animl Fcility Unit, Fculty of Science, Prince of Songkl University, Ht Yi, Thilnd. They were housed in n identicl wire-mesh-ottomed stinlesssteel cges contining six mice per cge nd mintined in n ir-conditioned room t 25 ± 2 C, 5 to 6% reltive humidity nd rtificil illumintion etween 6: nd 18: h. Commercil diets (C.P. Mice Feed, Chroen Phokphnd Group, Bngkok, Thilnd) nd drinking wter were provided d liitum. All procedures concerning niml tretments nd experimenttions in this study were reviewed nd pproved y the Institutionl Committee for Ethicl Use of Experimentl Animls t Prince of Songkl University Experimentl Design. In vivo ntioxidnt ctivity of R. tomentos leves extrct ws determined y using cron

3 Evidence-Bsed Complementry nd Alterntive Medicine 3 tetrchloride (CCl 4 )-induced oxidtive stress in the mouse model. The nimls were divided into six groups of six nimls ech. Before tretment, the mice were fsted overnight, ut with free ccess to wter. Group I served s control which received 1% DMSO (1 ml/kg/dy, intrgstric gvge (ig)) for 14 dys. Group II served s the toxin control nd received CCl 4 lone. Group III served s positive control nd received α-tocopherol (.1 g/kg/dy, ig) for 14 dys. Group IV to VI served s treted group nd received.2,.4, nd.8 g/kg ody weight of R. tomentos extrct in 1% DMSO (1 ml/kg/dy, ig), respectively, for 14 dys. Except Group I, ll other groups received sucutneous injection of 3% CCl 4 in olive oil (1 ml/kg, once in ech 72 h of 14 dys). All nimls hd free ccess to diet nd drinking wter during the study. The nimls were scrificed on dy 15 y cervicl decpittion. Blood smples were collected from the hert puncture into n nticogulnt-contining tue to prevent the clot. The liver nd kidney were quickly hrvested nd weighed. Blood nd tissue smples were stored t 8 Cfor further use Assessment of the Extent of Oxidtive Stress in Blood nd Tissues. The level of TBARS nd GSH, the ctivities of totl SOD, CAT, nd GPx in lood nd tissues were quntified using commercilly ville kits, purchsed from Cymn Chemicl Compny, USA, following the mnufcturer s instructions. Briefly, Liver nd kidney were homogenized (1% homogente) in different ssy uffers s per requirement of the ssy, nd the homogentes were centrifuged t 1, g for15mint4 C(SorvllRC5C PLUS centrifuge, USA). The otined superntnts collected to determine the levels of TBARS nd ntioxidnts ccording to the kit protocol. All the ssys were performed y using micro titer plte photometer (BioTek Instruments, Inc., USA) Sttisticl Anlysis. All the vlues re represented s men ± SEM. Dt for in vitro experimentswerenlyzed y liner regression nlysis nd t-test. Dt on iochemicl investigtions of in vivo experiments were nlyzed y onewy nlysis of vrince (ANOVA), nd the group mens were compred y dunnet s multiple rnge Test (DMRT). A proility of P<.5 ws considered s significnt. 3. Results nd Discussion 3.1. In Vitro Antioxidnt Activity Lipid Peroxidtion Inhiitory Assy. The inhiitory effect of cetone extrct from R. tomentos leves on FeCl 2 - scoric cid-induced mlondildehyde (MDA) production in liposome system is represented in Figure 1. Lipid peroxidtive degrdtion of the iomemrne is one of the principl mechnisms for the genertion of free rdicls [2]. The level of lipid peroxidtion ws determined s thiorituric cid rective sustnces, which re minly ldehydes nd 99% TBARS is mlondildehyde [21]. The rte of inhiition of MDA formtion incresed s the concentrtion of extrct mm of gllic cid equivlence Concentrtion of extrct (µg/ml) Figure 1: Inhiition of lipid peroxidtion t different concentrtionsof R. tomentos extrct. The vlues re expressed s gllic cid equivlence (men ± SEM; r 2 =.996, P<.1). n = 6pirs. mm of gllic cid nd ellgic cid equivlence/mg EAE R. tomentos extrct GAE Figure 2: Ferric ion reducing power of R. tomentos extrct t 1 mg/ml concentrtion. Vlues re expressed s gllic cid (GAE), nd ellgic cid (EAE) equivlence (men ± SEM; r 2 =.969, P<.4). n = 2 in triplictes. incresed. R. tomentos extrct ws le to significntly inhiit the genertion of lipid peroxides (P <.1). The lipid peroxidtion inhiition cpcity of the extrct ws equl to.93 ±.7 mm gllic cid, t 1 μg/ml Ferric Reducing Antioxidnt Power. The reducing cpcity of compound my serve s significnt indictor of its potentil ntioxidnt ctivity [22]. For the mesurement of reductive ility of R. tomentos extrct, the trnsformtion of Fe 3+ -Fe 2+ ws investigted in its presence. The extrct showed rpid nd incresed tendency to reduce Fe 3+ -Fe 2+ s equl s 1.8 ± 1.12 mm gllic cid nd 3.5 ± 5.22mMellgiccidcls, respectively, t1mg/ml (Figure 2). The extrct exhiited strong reducing ility of 2.7-fold nd 3.-fold higher thn tht of gllic cid nd ellgic cid, respectively Chelting Activity on Ferrous Ions. Trnsition metl ions such s ferrous ions re known to ctlyze the formtion of free rdicls, nd minority of metl ions could ccelerte

4 4 Evidence-Bsed Complementry nd Alterntive Medicine Tretment groups Tle 1: Antioxidnts sttus in the lood of mice in ech group. Level of nonenzymic ntioxidnt (Totl glutthione, μm/ml) Superoxide dismutse Level of ntioxidnt enzymes (U/mL) Ctlse Glutthione peroxidse 1% DMSO 4.12 ± ±.21 c 1.94 ±.87 c ±.47 c 3% CCl ± ± ± ± 1.2 α-tocopherol + CCl ±.82 c 4.44 ±.41 c 1.14 ±.78 c ±.99 Extrct(.2g/kg)+CCl ± ± ±.46 c 7.64 ± 1.71 Extrct(.4g/kg)+CCl ± ± ±.41 c 9.46 ±.24 Extrct(.8g/kg)+CCl ± ±.36 c 1.49 ±.48 c 1.64 ± 1.13 c Vlues re men ± SEM; P<.1 versus 1% DMSO-treted group (n = 6); P<.1 versus CCl 4 -treted group (n = 6); c P<.5 versus CCl 4 -treted group (n = 6). the lipid peroxidtion [23] y decomposing lipid hydroperoxides into peroxyl nd lkoxyl rdicls tht cn themselves strct hydrogen nd perpetute the chin rection of lipid peroxidtion [24]. Ferrozine, competitive metlion chelte, cn quntittively form complexes with Fe 2+. In the presence of chelting gents, the complex formtion is disrupted; with the result, the red colour of the complex is decresed. Mesurement of colour reduction, therefore, llows estimtion of the chelting ctivity of the coexisting chelte [25]. Acetone extrct from R. tomentos nd EDTA ( potent metl ion-chelting gent) interfered with the formtion of ferrous-ferrozine complex, suggesting tht they re cple to cpture ferrous ion efore ferrozine. Liner decrese in the sornce of Fe 2+ -ferrozine complex ws oserved, with the increse of plnt extrct concentrtion (Figure 3). The metl ion chelting ctivity of R. tomentos extrct is equl to.96 ±.1 mm EDTA, t 1 μg/ml. The ntioxidnt ctivity of puttive ntioxidnts hs een ttriuted to vrious mechnisms, mong which re prevention of chin initition, inding of trnsition metl ion ctlysts, decomposition of peroxides, prevention of continued hydrogen strction, reductive cpcity, nd rdicl scvenging cpility [26]. The rdicl scvenging cpility of the extrct my e due to the synergistic effect of the phytochemicls present in it In Vivo Antioxidnt Activity. Preliminry results from in vivo cute toxicity test elucidted tht the cetone extrct of R. tomentos is sfe when orlly dministered t concentrtion of 2 g/kg ody weight. In vivo ntioxidnt ctivity of R. tomentos lefextrctwscrriedouttdosesof.2,.4, nd.8 g/kg ody weight in mice induced with CCl 4. Cron tetrchloride is well-estlished heptotoxin nd used y vrious reserchers, for experimentl induction of oxidtive stress [27]. CCl 4 toxicity depends on the reductive dehlogention of CCl 4 ctlyzed y Cytochrome P 45 in endoplsmic reticulum leding to the genertion of n unstle complex trichloromethyl (CCl 3 ) rdicl, which initites cscde of free rdicl rections resulting in n increse of lipid peroxidtion nd reduction in some enzyme ctivities [28]. A numer of recent reports clerly demonstrted tht in ddition to heptic prolems, CCl 4 lso cuses disorders in kidneys, lungs, testis, nd rin s well s in lood y mm of EDTA equivlence Concentrtion of extrct (µg/ml) Figure 3: Metl ion (Fe 2+ ) chelting ctivity t different concentrtions of R. tomentos extrct. Vlues re expressed s EDTA equivlence (men ± SEM; r 2 =.9714, P<.1). n = 6pirs. generting free rdicls [29, 3]. Lipid peroxidtion nd levels of endogenous ntioxidnts re commonly used index to ssess the oxidtive stress. GSH, SOD, CAT, nd GPx from the primry tem of defense ginst rective oxygen species [31]. In vivo ntioxidnt ctivity of the extrct ws determined y nlyzing the levels of TBARS nd endogenous ntioxidnts in the lood, liver, nd kidneys of experimentl nimls. MDA is the end product of lipid peroxidtion in memrne ftty cids representing oxidtive dmge cused y free rdicls resulting in structurl modifiction of memrne with the relese of cell nd orgnelle contents, loss of essentil ftty cids in the production of cytosolic peroxide products [32]. The tripeptide, GSH, is the most importnt cellulr defense mechnism tht exists in the cell. The ility of the cell to regenerte GSH is n importnt fctor in the efficiency of mnging oxidtive insults. A depletion of intrcellulr GSH hs een reported during oxidtive stress conditions, where there is n increse in ROS [33]. In the present investigtion, the dministrtion of CCl 4 to the nimls resulted in elevtion of TBARS y out 4.11 folds (lood), 1.62 folds (liver), nd 2.58 folds (kidney) when compred to 1% DMSO treted nimls (Figure 4). In contrst to the elevtion of TBARS, significnt depletion of totl GSH levels y 3.5 folds (lood, P <.1), nd

5 Evidence-Bsed Complementry nd Alterntive Medicine folds (liver, P <.5) ws oserved (Tles 1, nd 2). However, no significnt decrese in GSH levels ws oserved in the kidney (Tle 3). Tretment of mice with R. tomentos leves extrct for 14 dys, dose dependently restored the TBARS levels nd GSH contents in lood, liver, nd kidney smples, comprle to those in 1% DMSO treted nimls. Similrly, α-tocopherol (1mg/kg ody weight) dministrtion lso prevented the formtion of TBARS nd mintined the GSH levels. Free rdicl scvenging enzymes such s SOD, CAT, nd GPx re the cellulr defense enzymes ginst oxidtive injury, decomposing superoxide, nd peroxide efore their interction to form the more rective hydroxyl rdicls [34]. The ctivities of SOD, CAT, nd GPx in lood, liver, nd kidneys of ll experimentl mice were represented in Tles 1, 2, nd 3, respectively. Severl in vivo studies hve demonstrted tht exposure to CCl 4 results in greter production of rective oxygen species leding to imlnce in the oxidnt/ntioxidnt sttus; hve een mnifested s lipid peroxidtion nd protein oxidtion, nd lso reported tht medicinl plnt extrcts re le to provide protection ginst the deleterious effects cused y free rdicls [35, 36]. In ll orgns of CCl 4 dministrted mice, significnt depletion in CAT nd GPx enzyme ctivities ws oserved, compred with 1% DMSO treted nimls. A significnt decrese in SOD ctivity ws oserved in the lood of CCl 4 - treted nimls, ut the decrese ws not significnt in liver nd kidney tissues. R. tomentos extrct exhiited protective effects ginst CCl 4 -induced decrese in SOD, CAT nd GPx enzyme ctivities in lood, liver, nd kidneys (Tles 1, 2, nd 3). At the lower doses (.2, nd.4 g/kg ody weight), the recovery in enzyme ctivities ws not significnt, while with higher dose of extrct (.8 g/kg ody weight), the recovery of enzyme ctivities were significnt (P <.5) when compred to the vlues otined from 1% DMSO treted mice. Alphtocopherol, tretment to the CCl 4 -injected mice, prevented the chnge in SOD, CAT, nd GPx ctivities of ll smples (Tles 1, 2, nd 3). The protective effect of R. tomentos t the dose of.8 g/kg ody weight, ginst CCl 4 induced oxidtive dmge ws similr to the effect of α-tocopherol (.1 g/kg ody weight). From the results of the present study, it ws oserved tht the order of the tissues tht re ffected y the oxidtive stress of CCl 4 on its iomolecules ws, lood > liver > kidney. The elevtion of lipid peroxidtion nd depletion of endogenous ntioxidnts in CCl 4 induced mice my e due to the free rdicls generted during the iotrnsformtion of CCl 4. Other workers hve reported tht R. tomentos t.4 g/kg ody weight cn effectively prevent the cetic cid-induced gstric ulcers vi its ntioxidnt ctivity [37]. It is suggested tht, R. tomentos extrct hs the cpcity to inhiit the effect of prooxidnts/oxidnts y reducing them nd preventing the initition of lipid peroxidtion chin rection. Considerle mount of reserch reports highlights the role of the polyphenolic constituents in the higher plnts s free rdicl scvenging gents [38 4]. We hve recently reported new flvellgic cid nd phloroglucinol (rhodomyrtosone I) from n cetone extrct of R. tomentos leves [41], in ddition to previous nm of MDA/mL nm of MDA/mg tissue nm of MDA/mg tissue c c CCl 4 D1 D2 D3 +CCl 4 () CCl 4 D1 D2 D3 +CCl 4 () c CCl 4 D1 D2 D3 +CCl 4 (c) Figure 4: Levels of thiorituric cid recting sustnces (TBARS) in lood (), liver (), nd kidneys (c) of mice in ech group. TBARS ws mesured s nm of mlondildehyde formed. Ech dt r represents men of six replictes in ech group ± SEM. Mens hving sme lphets on ech r do not differ (P.5) from ech other. Negtive control group: receive 1% DMSO (White column), toxin control group: receive 3% CCl 4 (Light gry column), positive control group: receive α-tocopherol + CCl 4 (Hevy gry column), nd extrct treted groups: received three different concentrtions of R. tomentos extrct (.2,.4, nd.8 g/kg) + CCl 4 (Blck column).

6 6 Evidence-Bsed Complementry nd Alterntive Medicine Tretment groups Tle 2: Antioxidnt sttus in the liver of mice in ech group. Level of nonenzymic ntioxidnt (Totl glutthione, μm/mgtissue) Superoxide dismutse Level of ntioxidnt enzymes (U/mg tissue) Ctlse Glutthione peroxidse 1% DMSO ± ± ±.31 c 2.65 ±.41 3% CCl ± ± ±.2.38 ±.6 α-tocopherol + CCl ± ± ±.49 c 2.51 ±.3 Extrct(.2g/kg)+CCl ± ± ± ±.18 Extrct(.4g/kg)+CCl ± ± ±.26 c 1.27 ±.23 Extrct(.8g/kg)+CCl ± ± ±.18 c 2.22 ±.63 Vlues re men ± SEM; P<.1 versus 1% DMSO-treted group (n = 6); P<.1 versus CCl 4 -treted group (n = 6); c P<.5 versus CCl 4 -treted group (n = 6). Tretment groups Tle 3: Antioxidnt sttus in the kidney of mice in ech group. Level of nonenzymic ntioxidnt (Totl glutthione; μm/mgtissue) Superoxide dismutse Level of ntioxidnt enzymes (U/mg tissue) Ctlse Glutthione peroxidse 1% DMSO 4.89 ± ± ± ± % CCl ± ± ±.1.31 ±.2 α-tocopherol + CCl ± ± ±.3 2. ±.26 Extrct(.2g/kg)+CCl ± ± ± ±.16 Extrct(.4g/kg)+CCl ± ± ± ±.19 Extrct(.8g/kg)+CCl ± ± ± ±.22 Vlues re men ± SEM; P<.1 versus 1% DMSO-treted group (n = 6); P<.1 versus CCl 4 -treted group (n = 6). reports on cylphloroglucinols such s rhodomyrtosones A-D, rhodomyrtone, nd some other known polyphenolic constituents including comretol, 3,3,4-tri-O-methylellgic cid, (6R,7E,9R)-9-hydroxy-4,7-megstigmdien-3-one nd α-tocopherol [42]. The presence of the identified phytochemicls my e responsile for the potent ntioxidnt ctivity of R. tomentos lef extrct. 4. Conclusion Both the in vitro nd the in vivo results of the study indicted tht R. tomentos extrct cn ct s potent ntioxidnt. Some of the ethnomedicl nd reported iologicl ctivities of this plnt re due its ntioxidnt property. Hence, it suggests tht R. tomentos extrct cn e employed s n ntioxidnt supplement in food products to prevent oxidtion of food nd lso s lterntive ntioxidnt therpeutics in the phrmcologicl industry to prevent the free rdicl dmge of iomolecules tht occurs in numerous humn diseses. However, further investigtions re needed to evlute the toxicity nd ntioxidnt ctivity of ech individul phytochemicl of the extrct to develop it into full-fledged ntioxidnt supplement/therpeutics. Authors Contriution S. P. Vorvuthikunchi: designed the reserch nd revised the pper. G. Lvny: conducted the reserch, nlyzed the dt, nd drfted the pper. N. H. Towtn: helped with portions of reserch nd revised the pper. Conflict of Interests The uthors declre tht there is no conflict of interests. None of the uthors hd ny finncil or personl interests in ny compny or orgniztion sponsoring the reserch currently or t the time of doing the reserch. Acknowledgments This work ws supported y the Higher Eduction Reserch Promotion nd Ntionl Reserch University project, Thilnd s Office of the Higher Eduction Commission. References [1] J. G. Scndlios, The rise of ROS, Trends in Biochemicl Sciences, vol. 27, no. 9, pp , 22. [2] J.M.Mtés, Effects of ntioxidnt enzymes in the moleculr control of rective oxygen species toxicology, Toxicology, vol. 153, no. 1 3, pp , 2. [3] A.Kdri,Z.Zri,I.B.Choetl., Chemiclcomposition nd ntioxidnt ctivity of Mrruium vulgre L. essentil oil from tunisi, Africn Journl of Biotechnology, vol. 1, no. 19, pp , 211. [4] A. Winoti, T. Wright, nd J. A. Goolsy, Herivores in Thilnd on Rhodomyrtus tomentos (Myrtcee), n invsive

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9 MEDIATORS of INFLAMMATION The Scientific World Journl Volume 214 Gstroenterology Reserch nd Prctice Volume 214 Journl of Dietes Reserch Volume Volume Volume 214 Interntionl Journl of Journl of Endocrinology Immunology Reserch Disese Mrkers Volume 214 Volume 214 Sumit your mnuscripts t BioMed Reserch Interntionl PPAR Reserch Volume 214 Volume 214 Journl of Oesity Journl of Ophthlmology Volume 214 Evidence-Bsed Complementry nd Alterntive Medicine Stem Cells Interntionl Volume Volume 214 Journl of Oncology Volume Volume 214 Prkinson s Disese Computtionl nd Mthemticl Methods in Medicine Volume 214 AIDS Behviourl Neurology Reserch nd Tretment Volume Volume Volume 214 Oxidtive Medicine nd Cellulr Longevity Volume 214