antimicrobial, antioxidant and anti-inflammatory activity of young shoots of the smoke tree, ...

* CorrespFaculty ofSerbia.E-mail: mi

PHYTOTHERAPY RESEARCHPhytother. Res. (2012)Published online in Wiley Online Library(wileyonlinelibrary.com) DOI: 10.1002/ptr.4919

Copyright

Antimicrobial, antioxidant and anti-inflammatoryactivity of young shoots of the smoke tree,Cotinus coggygria Scop

Mirjana Mar�cetić,1* Dragana Bo�zić,2 Marina Milenković,2 Nevenka Male�sević,3Sini�sa Radulović3 and Nada Kova�cević11Department of Pharmacognosy, Faculty of PharmacyUniversity of Belgrade, V. Stepe 450, 11221 Belgrade, Serbia2Department of Microbiology and Immunology, Faculty of Pharmacy University of Belgrade, Vojvode Stepe 450, 11221 Belgrade, Serbia3Institute of Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia

In this study, we investigated the antimicrobial activity of the young shoots of the smoke tree, Cotinus coggygriaScop., Anacardiaceae. The acetone extract and the derived ethyl acetate fraction effectively inhibited the growthof Gram-positive and Gram-negative bacteria (MIC 25–200mg/ml), while the chloroform fraction showed pro-nounced activity against the yeastCandida albicans (MIC 3.12mg/ml). The ethyl acetate fraction exhibited a signifi-cant ferric-reducing ability (10.7mmol Fe2+/g extract), a very highDPPH radical scavenging activity (SC50=1.7mg/ml)and inhibition of lipid peroxidation (IC50=41.8mg/ml). High amounts of total phenolics (929.8mg/g), tannins(833.8mg/g) and flavonoids (35.5mg/g) were determined in the ethyl acetate fraction, which also exerted significantanti-inflammatory (76.7%) and cytotoxic effects (IC50=15.6mg/ml). Copyright © 2012 John Wiley & Sons, Ltd.

Keywords: Cotinus coggygria; young shoots; antimicrobial activity; antioxidant activity; anti-inflammatory activity.

INTRODUCTION

Cotinus coggygria Scop. (Rhus cotinus L.), Anacardia-ceae (smoke tree) is a shrub found in south and south-east Europe. In the traditional medicine of differentcountries, C. coggygria has been used for its antiseptic,anti-inflammatory and antihaemorrhagic properties. Itis mainly applied to treat diarrhoea, in wound healingand for different skin disorders. In Serbian folk medi-cine, decoction of the bark has also been used to treatcancer (Ki�sgeci, 2002; Demirci et al., 2003; Tzakouet al., 2005).Cotinus coggygria contains an essential oil which

mainly consists of monoterpenes (Demirci et al., 2003;Tzakou et al., 2005; Novaković et al., 2007), a high amountof hydrolysable tannins, gallic acid, methyl gallate, penta-galloyl glucose (Westenburg et al., 2000) and flavonoids,glycosides of myricetin, quercetin and kaempferol(Buziashvili et al., 1973). Three aurones (disulfuretin, sul-furetin and sulfurein) possessing antioxidant activity(DPPH radical scavenging activity) were isolated fromC. coggygria (Westenburg et al., 2000). A methanolextract of the leaves and flowers exhibited antioxidantand cytotoxic activities (Šavikin et al., 2009). The anti-microbial activity of the essential oil of C. coggygria fromTurkey was not significant (Demirci et al., 2003), whilethe essential oil found in the leaves of plants growing inSerbia inhibited the growth of Gram-positive bacteriaand fungi (Novaković et al., 2007).

ondence to: Mirjana Mar�cetić, Department of Pharmacognosy,Pharmacy University of Belgrade, V. Stepe 450, 11221 Belgrade,

[email protected]

© 2012 John Wiley & Sons, Ltd.

In view of growing microbial resistance, plant second-ary metabolites are proving to be promising candidatesas potential new antimicrobial agents. The aim of ourstudy was to evaluate the antimicrobial activity of anacetone extract of the young shoots of C. coggygriaand its derived fractions, as this has not been investi-gated so far. Reactive oxygen species (ROS) are pro-duced during aerobic metabolism of mammalian cells.When the balance between ROS production and anorganism’s antioxidant defences (endogenous and nutri-tional) is disturbed, oxidative stress develops (Halliwell,2006). Excessive oxidative stress is known to present arisk to human health and correlates closely to degenera-tive diseases, chronic inflammatory processes (Roessneret al., 2008), cardiovascular diseases and cancer (Ziechet al., 2012). The antioxidant activities of the acetoneextract of young shoots of C. coggygria and of thederived chloroform, ethyl acetate and water fractionswere researched. It has been established that ROScause cell and tissue damage and act as signallingmolecules in the processes of inflammation and carcino-genesis (Santangelo et al., 2007). In this work, the anti-inflammatory activity of the C. coggygria ethyl acetatefraction was assessed in an in vivo model of ratpaw oedema. Its potential cytotoxic properties werealso examined.

MATERIALS AND METHODS

Plant material and extraction procedure. Young shoots,which included leaves and branches ofCotinus coggygria,were collected near the Resavica River in central Serbia,during July 2004. Identification was confirmed byProf. Dr Branislava Laku�sić from the Department of

Received 26 July 2011Revised 26 November 2012

Accepted 30 November 2012

Table 1. Contents of total polyphenols, tannins and flavonoids inC. coggygria acetone extract and fractions.

Extract/Fraction

Yield Total phenolics Tannins Flavonoids

(%) (GAE) (mg/g) (GAE) (mg/g) (mg/g)

Acetone 42.5 518.4�0.6 494.4�0.2 23.2�0.4Chloroform 7.3 171.7�0.5 91.2�0.3 25.6�0.5Ethyl acetate 53.0 929.8�0.8 833.8�0.8 35.5�0.2Water 39.7 77.5�0.1 47.0�0.2 2.6�0.0

M. MAR�CETIĆ ET AL.

Botany, Faculty of Pharmacy, University of Belgrade,Serbia. A voucher specimen has been deposited there.Air-dried and groundmaterial was extractedwith acetone(70%V/V) at room temperature. The solvent was evapo-rated under reduced pressure below 40�C. The solidacetone extract was suspended in water and partitionedwith chloroform and ethyl acetate to yield chloroform,ethyl acetate and water fractions. The solvents wereevaporated under reduced pressure below 40�C.

Total phenolic, tannin and flavonoid contents. The totalphenolic and tannin contents in the C. coggygria acetoneextract and its derived fractions were analyzed spectro-photometrically by the Folin-Ciocalteu assay. Theremoval of tannin was achieved with insoluble polyvinyl-pyrrolidone which binds tannins (Makkar et al., 2000).Total phenolic and tannin contents were expressed asmg of gallic acid equivalents/g extract (GAE). The totalflavonoid content was determined spectrophotometric-ally, based on the formation of a flavonoid-aluminiumcomplex, as described in the monograph for birch leaf,Betulae folium, (European Pharmacopoeia, 2007).HPLC identification of the phenolic constituents was

performed with a Zorbax Eclipse XDB-C18 analyticalcolumn and a photodiode array detector. Samples wereeluted with a gradient two-phase system (Tomić et al.,2009). Identification of components was performed bycomparing the UV spectra and peak retention timeswith the recorded UV spectra and peak retention timesof representative standards.

Antimicrobial activity. The antimicrobial activity of C.coggygria extracts was examined by the broth microdilu-tion method (NCCLS, 2001) against the eight differentlaboratory control strains of bacteria and two strains ofyeast listed in Table 1. Minimal inhibitory concentration(MIC) and minimal bactericidal or fungicidal concentra-tions (MBC) were determined according to the NationalCommittee for Clinical Laboratory Standards (NCCLS,2001). All tests were performed in Müeller-Hintonbroth for bacterial strains and in Sabouraud dextrosebroth for C. albicans. Overnight broth cultures of eachstrain were prepared at a final concentration of 2� 106

CFU/ml for bacteria, and at 2� 105CFU/ml for yeast,in a 96-well microtiter plate. Extracts were investigatedin the range 3.12–400 mg/ml. Triphenyl tetrazoliumchloride was added to the culture medium as a growthindicator (0.05%). Microbial growth was determinedafter incubation at 37�C for 24 h for bacteria, and afterincubation for 48 h at 26�C for fungi. The MIC is definedas the lowest concentration of the extract at which themicroorganism does not demonstrate visible growth.MBC is defined as the lowest concentration of theextract at which inoculated microorganisms are com-pletely killed. To determine the MBC, broth was takenfrom each well (10ml) and inoculated in Müeller-Hintonagar for 24h at 37�C for bacterial strains, and in Sabourauddextrose agar for 48h at 26�C for the fungi. All determina-tions were performed in duplicate; two positive growthcontrols were included.

Ferric reducing antioxidant power. The total antioxidantpotential was determined by the ferric reducing antioxi-dant power (FRAP) assay, based on the reduction of theferric ion (Fe3+) to a ferrous ion (Fe2+) and the formationof a blue complex (Fe2+-2,4,6-tripyridyl-s-triazine), which

Copyright © 2012 John Wiley & Sons, Ltd.

increases the absorption at 593nm (Benzie and Strain,1996; Szőllősi and Varga Szőllősi, 2002). The amount ofreduced Fe2+-TPTZ complex was calculated from anequation of regression line constructed from the absorb-ance of water solutions of FeSO4� 7H2O (0.1–1.0mmol)under experimental conditions. Results are expressed asmmol Fe2+/g extract.

DPPH radical scavenging activity.DPPH radical scaven-ging activity is based on the reduction of the free radicalDPPH• (2,2-diphenyl-1-picrylhydrazyl) and the decreaseof absorbance at 517 nm (Bondet et al., 1997). Ascorbicacid and gallic acid served as the control.

Inhibition of lipid peroxidation. Lipid peroxidation (LP)on liposomes was induced by an Fe2+/ascorbate systemand measured spectrophotometrically by the thiobarbitu-ric acid (TBA) test, based on the determination of theabsorbance of a product between malondialdehyde andTBA (Afanas’ev et al., 1989). Gallic acid and tannin(polymer of gallic acid molecules and glucose) were usedas reference control.

Anti-inflammatory activity. The carrageenan-induced ratpaw oedema test served to examine the anti-inflammatoryactivity (Oyanagui and Sato, 1991). All experimental pro-cedures and protocols conformed to institutional guide-lines for the care and use of animals in research No 2/09(Ethics Committee of the Faculty of Pharmacy, Belgrade,Serbia). Experimental groups consisted of six adult maleWistar rats, weighing 150–200 g. The ethyl acetate extractwas orally administered in doses of 50 and 100mg/kg.Indomethacin was used as a reference in a dose of 8mg/kg.

Cytotoxic activity on HeLa cells. The cytotoxicity of theC. coggygria ethyl acetate extract was determined usingthe Kenacid Blue R dye binding method (Clothier,1995) on HeLa (human cervix carcinoma) cells. Absorb-ance was recorded on an enzyme-linked immunosorb-ent assay (ELISA) reader at a wavelength of 570 nm.The IC50 (mg/ml) was defined as the concentration ofextract that produced 50% inhibition of cell survival. Itwas determined from cell survival diagrams.

Statistical analysis. All determinations were made in trip-licate. Data were expressed as means� SD. Statisticalanalysis was performed by the Mann–Whitney U-test.Differences were taken to be statistically significantwhen P< 0.05. The IC50, SC50 were calculated from thedose–response curves obtained by plotting the percentageof inhibition versus the concentrations.

Phytother. Res. (2012)

ANTIMICROBIAL ACTIVITY OF SMOKE TREE

RESULTS AND DISCUSSION

Phenolic compounds have multiple biological effects, suchas anti-atherogenic, antioxidant, anti-inflammatory, cardi-oprotective, antimicrobial, anticarcinogenic and neuro-protective (Manach et al., 2005; Middleton et al., 2000).The contents of total phenolics, tannins and flavonoidsof the C. coggygria extract and its derived fractions aregiven in Table 1. The acetone extract contained a highamount of total phenolics, tannins and flavonoids. Previ-ous investigations (Nićiforović et al., 2010) showed thatthe C. coggygria methanol extract had a very high quan-tity of total phenolics (413mg/g), which is similar to ourresults for the acetone extract (518.4mg/g). The highestcontent of total phenolics was determined in the semi-polar ethyl acetate fraction and the lowest in the polarwater fraction. Analogous with this, the amounts oftannin and flavonoids in the ethyl acetate fraction weremuch higher than in other fractions.The acetone extract of C. coggygria is characterised

by the presence of the phenolic acid gallic acid, gallicacid derivatives and flavonol kaempferol-3-O-glucoside.In the ethyl acetate fraction, gallic acid, gallic acid deri-vatives, kaempferol-3-O-glucoside and the flavonesluteolin-7-O-glucoside, luteolin-8C-glucoside (orientin),apigenin glycoside and apigenin were detected.

Antimicrobial activity

Microbial antibiotic resistance is increasing worldwideand can be demonstrated against all classes of antibiotics.Moreover, multi-drug-resistant strains have emerged inmany species of pathogenic microorganisms (Orhanet al., 2010). Consequently, there is a growing interest innew sources of antimicrobial agents. Plant secondarymetabolites such as terpenoids, phenolic compounds,alkaloids and lectins exert an antimicrobial effect(Cowan, 1999). The antimicrobial activity, expressedas the MIC of the acetone extract and the fractionsobtained from young shoots of C. coggygria were in therange 3.1–200mg/ml (Table 2). The antimicrobial effect ofan extract is considered to be significant if the MIC valuesof the investigated extract are below 100mg/ml (Ríos andRecio, 2005). The acetone extract and ethyl acetate frac-tion were very active against S. aureus (MIC 25mg/ml), abacteria that causes a range of illnesses from minor skin

Table 2. Minimal inhibitory concentration (MIC) of young shoots of C

Microorganisms A

Staphylococcus aureus ATCC 25923 25Staphylococcus epidermidis ATCC 12228 25Micrococcus luteus ATCC 9341 50Enterococcus faecalis ATCC 29212 100Bacillus subtilis ATCC6633 100Pseudomonas aeruginosa ATCC27853 100Escherichia coli ATCC25922 100Klebsiella pneumoniae NCIMB 9111 200Candida albicans ATCC 10259 50Candida albicans ATCC 24433 6.25

A: acetone extract; C: chloroform fraction; E: ethyl acetate fraction; W:


infections to life-threatening diseases (Yu et al., 2008).The acetone extract also pronouncedly inhibited thegrowth of the Gram-positive bacteria S. epidermidis(MIC 25mg/ml), whereas the ethyl acetate fraction wasactive against B. subtilis (MIC 25mg/ml). It is importantto stress the antimicrobial effect of the ethyl acetate frac-tion against the Gram-negative bacteria K. pneumoniae(MIC 50mg/ml) and E. coli (MIC 50mg/ml), since thesebacteria are usually resistant to commonly used antibio-tics. Plant secondary metabolites often show activityagainst Gram-positive bacteria, but not against Gram-negative bacteria or yeast. Gram-negative bacteria havean effective permeability barrier that restricts the pene-tration of compounds (Tegos et al., 2002). The ethylacetate fraction contained a high amount of total pheno-lics, tannins and flavonoids. The antimicrobial effect ofphenolic compounds is based on their ability to inactivatemicrobial adhesions, enzymes and cell envelope transportproteins (Cowan, 1999). Interestingly, the highest activitywas obtained with the non-polar chloroform fractionon the opportunistic yeast C. albicans ATCC 24433(MIC 3.1mg/ml). The chloroform fraction inhibited thegrowth of C. albicans ATCC 24433 more effectively thanthe control antifungal drug nystatin (6.2mg/ml), althoughthe acetone extract also exerted very high activity (MIC6.2mg/ml). The antimicrobial effect of the ethyl acetatefraction is probably due to its high phenolic content. Onthe other hand, the chloroform fraction, which has a loweramount of phenolics, markedly inhibited the growth ofC. albicans. The phenolic compounds are not only respon-sible for the antimicrobial activity of young shoots ofC. coggygria. This is in agreement with the previousdemonstration of the high antimicrobial effect of theessential oil from C. coggygria leaves on Gram-positivebacteria and fungi (Novaković et al., 2007). Our investiga-tion showed that, beside the Gram-positive bacteria andfungi, the ethyl acetate fraction also significantly inhibitedthe growth of resistant Gram-negative bacterial strains.

The MBC values of the investigated extract and frac-tions were higher than 200mg/ml for bacteria, and werein the range 100–200mg/ml for C. albicans.

Antioxidant activity

The antioxidant activity of phenolic compounds is basedon their ability to donate hydrogen atoms or electrons,

otinus coggygria acetone extract and its derived fractions

MIC (mg/ml)

C E W

100 25 100 0.5Amp

50 50 200 1Amp

100 50 100 0.5Amp

50 200 200 1Amp

50 25 100 2Amp

100 100 200 4Ami

200 50 200 2Ami

200 50 200 2Ami

100 50 50 3.1Nys

3.12 50 50 6.2Nys

water fraction; Amp: ampicillin; Ami: amikacin; Nys: nystatin


Figure 1. Inflammatory response in the rats with the carrageenan-induced paw oedema treated with the ethyl acetate fraction of C.coggygria. Results are expressed as mean�SD (n=6).


chelate metal cations and scavenge free radicals (Bondetet al., 1997; Afanas’ev et al., 1989). The ferric reducingpower of the young shoots of C. coggygria is presentedin Table 3. The acetone extract and ethyl acetate fractionexhibited high reducing power. The reducing ability of theethyl acetate fraction is comparable with the activity ofthe well-known antioxidant, ascorbic acid.The DPPH assay gives data about the reactivity of a

potential antioxidant with the stable free DPPH radical.It enables evaluation of the effectiveness of the examinedextracts to inhibit the production of reactive oxygen radi-cals. All the investigated extracts showed an ability toscavenge the DPPH radical (Table 4). The concentrationsof extracts required to achieve 50% scavenging of theinitial amount of DPPH (SC50) were in the range1.7–25.0mg/ml. The whole C. coggygria plant (Westenburget al., 2000; Nićiforović et al., 2010) and its leaves and flow-ers (Šavikin et al., 2009) have presented a high antioxidanteffectiveness. The acetone extract of the young shoots ofC. coggygria exhibited a more pronounced activity thanascorbic acid, and an effect that was similar to that ofgallic acid. The ethyl acetate fraction of C. coggygriaexhibited a very high DPPH radical scavenging activity(SC50 1.7mg/ml) that was higher than that of the controlantioxidants, ascorbic acid and gallic acid.The process of LP has an important role in oxidative

stress development. The peroxidation of unsaturatedlipids of the cell membrane leads to cell membrane dam-age, loss of membrane integrity and consequently to celland tissue damage (Halliwell, 2006). The investigated ex-tract and its derived fractions displayed dose-dependentantioxidant activities. The results of the LP assay weresimilar to the FRAP and DPPH assays. The ethyl acetatefraction exhibited the highest capability of inhibiting LP,while the water-soluble fraction was the weakest. At aconcentration of 400mg/ml, the water fraction inhibited50.3� 7.9% of LP, the chloroform fraction 56.1� 4.4%,the acetone extract 83.2� 3.7% and the ethyl acetate frac-tion 90.5� 6.3%. Gallic acid and tannin, which were usedas control antioxidants, inhibited 44.4� 6.4% and 73.6�2.2% of LP, respectively, at the same concentration. Theseresults also showed that the C. coggygria acetone extractand ethyl acetate fraction exhibited stronger inhibition ofLP than the individual constituents gallic acid and tannin.

Preliminary anti-inflammatory and cytotoxic activity

ROS damage cell membranes and DNA, causing poten-tially cancerous mutations. They are also implicated in

Table 3. Ferric reducing power, DPPH radical scavenging activity, inhiC. coggygria acetone extract and its derived fractions

Extract/Fraction

FRAP

mmol Fe2+/g extract SC

Acetone 8.4�0.2Chloroform nt 1Ethyl acetate 10.7�1.1Water 1.4�0.2 2Ascorbic acid 10.4�0.1Gallic acid ntTannin nt

nt-not tested


inflammatory processes (Roessner et al., 2008). Phenoliccompounds have an ability to inhibit the production ofROS or to neutralise radicals that are involved in the in-flammatory process as signalling molecules (Santangeloet al., 2007). The ethyl acetate fraction of the youngshoots of C. coggygria exhibited the highest antioxidanteffect. Therefore, the anti-inflammatory and cytotoxicactivities of this fraction were investigated. The ethylacetate fraction reduced carrageenan rat paw oedemain a dose-dependent manner (Fig. 1). Doses of 50mg/kgand 100mg/kg led to 46.5� 18.5% and 76.7� 0.0%reduction of the oedema, respectively, being statisticallyvery significant (p< 0.01). The anti-inflammatory effectof the ethyl acetate fraction at a dose of 100mg/kg wasmore pronounced than the activity of the anti-inflammatorydrug indomethacin (53.8� 14.6%) (p< 0.01). Phenoliccompounds inhibit cyclooxygenase-1 and cyclooxygenase-2,which are responsible for the production of theinflammatory mediators, prostaglandins (Yoon andBaek, 2005). Gallic acid decreases the release ofhistamine, a vasoactive mediator implicated in theacute phase of immediate hypersensitivity. It reducesthe elevated intracellular calcium levels and sup-presses pro-inflammatory cytokine production, lead-ing to the reduction of the inflammatory reaction(Kim et al., 2006). Kaempferol inhibits phospholipaseA2, the enzyme involved in the production of fattyacids, such as arachidonic acid. Apigenin restrainsthe production of nitric oxide and subsequently theconversion of arachidonic acid into prostaglandins(Yoon and Baek, 2005).

bition of lipid peroxidation and cytotoxic activity on HeLa cells of

DPPH LP Cytotoxic

50 (mg/ml) IC50 (mg/ml) IC50 (mg/ml)

3.5�0,2 51.3�3.9 nt3.1�0,8 147.4�5.8 nt1.7�0,2 41.8�7.0 15.6�0.85.0�0,6 280.2�3.7 nt4.1�0.1 nt nt3.5�0.1 817.9�6.4 10.0�0.5

nt 73.6�2.2 17.3�6.9


ANTIMICROBIAL ACTIVITY OF SMOKE TREE

The ethyl acetate fraction of the acetone extract ofyoung shoots of C. coggygria exerted a strong dose-dependent cytotoxic activity on HeLa cells. The cytotoxiceffect of the ethyl acetate fraction was more pronouncedthan the activity of tannin, but weaker than the cytotox-icity of gallic acid.The results obtained in this study suggest that the

young shoots of C. coggygria possess a broad-spectrumof activity against several tested microbial strains. Theethyl acetate fraction, which contained the highestamount of total phenolics, tannin and flavonoids, effect-ively inhibited the growth of Gram-positive and Gram-negative bacteria. A very high antioxidant activity wasobserved in the ethyl acetate fraction, which also showedsignificant anti-inflammatory and cytotoxic effects. It hasbeen demonstrated that the gallate structure in phenoliccompounds is essential for anti-inflammatory activity(Santangelo et al., 2007). The ethyl acetate fraction ofC. coggygria contains very high amounts of tannins con-sisting of gallic acid and gallic acid derivatives. The gallatestructure also had a crucial role in the inhibition of thematrix-proteases, leukocyte elastase and gelatinases thatare involved in inflammation and cancer invasion (Sartoret al., 2002). The significant anti-inflammatory activity ofthe ethyl acetate fraction is probably based on its highantioxidant activity. We also demonstrated that the ethylacetate fraction exerted a cytotoxic effect. Galati and


O’Brien (2004) showed that flavonoids can act as antiox-idants or as pro-oxidants in the presence of transitionmetals. The phenol B ring-containing flavonoids areoxidised by peroxidases to phenoxyl radicals, which canlead to electrophilic metabolites that bind to cellularmacromolecules and generate ROS. Apoptotic DNAfragmentation mediated by ROS may be an importantmechanism of phenolic cytotoxic activity, rather than itsantioxidant activity (Galati and O’Brien, 2004). It canbe concluded that C. coggygria, and especially itsyoung shoots, is a valuable source of effective phenoliccompounds that could prevent oxidative stress-inducedcellular damage, suppress inflammatory reactions andprevent infections.

Acknowledgements

This work was financially supported by the Ministry of Education,Science and Technological Development of the Republic of Serbia,Project No. 173021.

Conflict of Interest

The authors declare that there is no conflict of interest.

REFERENCES

Afanas’ev IB, Dorozhko AI, Brodskii AV, Kostyuk VA, PotapovitchAI. 1989. Chelating and free radical scavenging mechanismsof inhibitory action of rutin and quercetin in lipid peroxidation.Biochem Pharmacol 38: 1763–1769.

Benzie IFF, Strain JJ. 1996. The Ferric Reducing Ability of Plasma(FRAP) as a Measure of "Antioxidant Power": The FRAPAssay. Anal Biochem 239: 70–76.

Bondet V, Brand-Williams W, Berset C. 1997. Kinetics andMechanisms of Antioxidant Activity using the DPPH˙ FreeRadical Method. Lebensm-Wiss u-Technol 30: 609–615.

Buziashvili IS, Komissarenko NF, Kolesnikov DG. 1973. Polyphenoliccompounds in the leaves of the Venetian sumac (Rhus coriaria)and sumac (Cotinus coggygria). Fenol’nye Soedin Ikh FiziolSvoistva, Mater Vses Simp Fenol’nym Soedin, 2nd: 159–162.

Clothier RH. 1995. The FRAME cytotoxicity test.MethodsMol Biol43: 109–118.

Cowan MM. 1999. Plant Products as Antimicrobial Agents. ClinMicrobiol Rev 12: 564–582.

Demirci B, Demirci F, Başer KHC. 2003. Composition of the essen-tial oil of Cotinus coggygria Scop. from Turkey. Flavour FragJ 18(1): 43–44.

European Pharmacopoeia. 2007. Sixth Edition. Ph. Eur. 6.0. Coun-cil of Europe: Strasbourg.

Galati G, O’Brien PJ. 2004. Potential toxicity of flavonoids and otherdietary phenolics: significance for their chemopreventive andanticancer properties. Free Radic Biol Med 37: 287–303.

Halliwell B. 2006. Reactive species and antioxidants. Redox biologyis a fundamental theme of aerobic life. Plant Physiol 141 (2):312–322.

Kim S-H, Jun C-D, Suk K, et al. 2006. Gallic Acid Inhibits HistamineRelease and Pro-inflammatory Cytokine Production in MastCells. Toxicol Sci 91(1): 123–131.

Ki�sgeci J. 2002. Lekovito bilje, gajenje, sakupljanje, upotreba. Parte-non: Beograd; 146.

Makkar HPS, Hagerman A, Harvey-Mueller I. 2000. Quantificationof Tannins in Tree Foliage – A Laboratory Manual. FAO/IAEA:Vienna.

Manach C, Mazur A, Scalbert A. 2005. Polyphenols and preventionof cardiovascular diseases. Curr Opin Lipidol 16(1): 77–84.

Middleton E, Kandaswami C, Theoharides TC. 2000. The effect ofplant flavonoids on mammalian cells: implication for inflamma-tion, heart disease and cancer. Pharmacol Rev 52(4): 673–751.

NCCLS. 2001. National Committee for Clinical LaboratoryStandards, Performance Standards for Antimicrobial Suscepti-bility Testing, 11th informational supplement, DocumentM100-S11. National Committee for Clinical Laboratory Standard:Wayne, PA.

NićiforovićN,MihailovićV,Ma�sković P, SolujićS, StojkovićA, PavlovićMuratspahićD. 2010. Antioxidant activity of selected plant spe-cies; potential new sources of natural antioxidants. Food ChemToxicol 48: 3125–3130.

Novaković M, Vu�cković I, Jana�cković P, et al. 2007. Chemicalcomposition, antibacterial and antifungal activity of the essen-tial oils of Cotinus coggygria from Serbia. J Serb Chem Soc 72(11): 1045–1051.

Orhan DD, Özçelik B, Özgen S, Ergun F. 2010. Antibacterial, antifun-gal, and antiviral activities of some flavonoids. Microbiol Res165(6): 496–504.

Oyanagui Y, Sato S. 1991. Inhibition by nilvadipine of ischemic and car-rageenan paw edema as well as of superoxide radical productionfrom neutrophils and xanthine oxidase. ArzneimittelforschungDrug Res 41(5): 469–474.

Ríos JL, Recio MC. 2005. Medicinal plants and antimicrobial activ-ity. J Ethnopharmacol 100: 80–84.

Roessner A, Kuester D, Malfertheiner P, Schneider-Stock R. 2008.Oxidative stress in ulcerative colitis-associated carcinogen-esis. Pathol Res Pract 204(7): 511–524.

Santangelo C, Varì R, Scazzocchio B, Di Benedetto R, Filesi C,Masella R. 2007. Polyphenols, intracellular signalling andinflammation. Ann Ist Super Sanita 43: 394–405.

Sartor L, Pezzato E, Dell’Aica I, Caniato R, Biggin S, Garbisa S.2002. Inhibition of matrix-proteases by polyphenols: chemicalinsights for anti-inflammatory and anti-invasion drug design.Biochem Pharmacol 64: 229–237.

Šavikin K, Zdunić G, Janković T, Stanojković T, Juranić Z, MenkovićN. 2009. In vitro cytotoxic and antioxidative activity ofCornus mas and Cotinus coggygria. Nat Prod Res 23(18):1731–1739.

Szőllősi R, Varga Szőllősi I. 2002. Total antioxidant power in somespecies of Labiatae (Adaptation of FRAP method). Act BiolSzeg 46(3-4): 125–127.

Tegos G, Stermitz FR, Lomovskaya O, Lewis K. 2002. Multidrugpump inhibitors uncover remarkable activity of plant antimicro-bials. Antimicrob Agents Chemother 46(10): 3133–3141.



Tomić A, Petrović S, Pavlović M, et al. 2009. Antimicrobial andantioxidant properties of methanol extracts of two Athamantaturbith subspecies. Pharm Biol 47(4): 314–319.

Tzakou O, Bazos I, Yannitsaros A. 2005. Essential oils of leaves,inflorescences and infructescenses of spontaneous Cotinuscoggygria Scop. from Greece. Flavour Frag J 20(5): 531–533.

Westenburg HE, Lee K-J, Lee SK, et al. 2000. Activity-guided iso-lation of antioxidative constituents of Cotinus coggygria. JNat Prod 63(12): 1696–1698.


Yoon J-H, Baek SJ. 2005. Molecular targets of dietary polyphenolswith anti-inflammatory properties.YonseiMed J46(5): 585–596.

Yu L, Xiang H, Fan J, et al. 2008. Global transcriptional responseof Staphylococcus aureus to Rhein, a Natural Plant Product.J Biotechnol 135: 304–308.

Ziech D, Anestopoulos I, Hanafi R, et al. 2012. Pleiotrophic effectsof natural products in ROS-induced carcinogenesis: The role ofplant-derived natural products in oral cancer chemopreven-tion. Cancer Lett 327: 16–25.


antimicrobial, antioxidant and anti-inflammatory activity of young shoots of the smoke tree, ...

Documents