Summary. Original Article. Full-text PDF: Postepy Hig Med Dosw (online), 2014; 68: e-issn

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1 Postepy Hig Me Dosw (online), 2014; 68: e-issn Originl Artile Reeive: Aepte: Pulishe: Authors Contriution: Stuy Design Dt Colletion Sttistil Anlysis Dt Interprettion Mnusript Preprtion Literture Serh Funs Colletion Biologil tivity n stility in vitro of polyphenoli extrts s potentil ietry supplements* Aktywność iologizn i stilność in vitro ekstrktów polifenolowyh jko potenjlnyh suplementów iety Pulin Strugł,,,,, JninGrielsk,,,, Deprtment of Physis n Biophysis, Wrołw University of Environmentl n Life Siene Introution: Mterils n methos: Results: Conlusion: Key wors: Summry In times of worsening iviliztion iseses the interest in nturl heling sustnes is on the inrese. To reue unwnte sie effets of mny syntheti rugs, it is resonle to introue to the ily iet foos rih in nturl ompouns of plnt origin tht re enefiil for helth. The purpose of the stuy ws to etermine the iologil tivity n stility of selete ethnol extrts of the fruit of hokeerry, lkurrnt, hwthorn, rosehip, quine n Jpnese quine s potentil nutreutils. Antioxint tivity of the extrts ws etermine in reltion to moel phospholipi memrnes (IC 50 ). Antiril tivity ws etermine in test with the ril (IC 50 ). Also the inhiition of enzymti (1-LOX) oxition of linolei i ws etermine t the eginning of the perio of storge of the extrts t room temperture n fter 12 months. After 12 months of storge the highest ntioxint stility ws shown y lkurrnt extrt (1.5% inrese in IC 50 ), the highest ntiril stility y quine extrt (1.0% reution in IC 50 ), n the highest stility of 1-LOX enzyme inhiition y hokeerry extrt (6.3% reution in inhiition t onentrtion of 8 μg ml -1 ). Jpnese quine extrt showe the strongest regenerting properties with respet to oxiize phospholipi memrnes n the highest ility to eliminte the free ril. It n e onlue tht the ethnol extrts of the fruits (in prtiulr lkurrnt, hokeerry n Jpnese quine) re potentil soure of ietry supplements of expete effetiveness in preventive tretment. polyphenoli fruit extrts ntioxints ntiril nti-inflmmtory tivities liposome memrne ietry supplements Full-text PDF: Wor ount: Tles: Figures: Referenes: *This work ws supporte y the NCN for reserh in the yers , grnt no. N N n prtly from funs of sttutory tivities of the Deprtment of Physis n Biophysis of Wrołw University of Environmentl n Life Sienes Postepy Hig Me Dosw (online), 2014; 68

2 Strugł P., Grielsk J. Biologil tivity n stility in vitro... Author s ress: Pulin Strugł MS, Deprtment of Physis n Biophysis, University of Environmentl n Life Siene, Norwi 25, Wrołw, Poln; e-mil: pulin.strugl@up.wro.pl Introution In reent yers there hs een lrge inrese in onsumer interest in the influene of nutrition on humn helth. There is inrese emn for foo n ietry supplements with enefiil effets on the oy. Current trens in foo tehnology len in the iretion of pling on the mrket mny new preprtions, ietry supplements ontining helth-supporting omponents. Their iologil properties re well oumente [44,45]. Compouns of the polyphenol group re rih soure of nturl sustnes of ntioxint hrter [20]. The literture reports tht nturl polyphenols reue the risk of iviliztion iseses suh s hypertension, oronry rtery isese, theroslerosis, n ner [8,33,42]. Reserh onute on extrts erive from fruits, vegetles n hers inites tht they my hve signifint role in the prevention n tretment of iseses. Their phrmologil effetiveness is often onnete with the pture of free rils, lipi oxition protetion n nti-inflmmtory tion [47]. There is lso growing interest in funtionl foos n foo itives, whih my ssist the efense mehnisms ginst iseses use y oxitive stress. The fruit of hokeerry, hwthorn, quine, Jpnese quine, lkurrnt n rosehip, originting from res of south-est Poln, whih so fr re reltively little ppreite y onsumers, ws selete for the stuy. Chokeerry n lkurrnt, rih in nthoynins, re vlule soure of iologilly tive ompouns. Consumption of this fruit uses, mong other effets, lowering of loo pressure [16]. Chokeerry shows ntimutgeni [14] n ntilotting tivity [31]. Also onfirme is the ility of lkurrnt to inhiit evelopment of the type A n B influenz virus in high temperture environment [21]. Reserh on the humn oloretl ner ell line Co-2 showe signifint reution in prolifertion of nerous ells in the presene of hokeerry n lkurrnt extrt [2,3,4]. Sientifi reports unerline the roer spets of helth enefiil effets of rosehip n hwthorn. Rosehip hs een known for yers in herl meiine n use in treting iseses of the liver, gll ler n kiney [17]. It positively ffets the mintenne of helthy n flexile joints, n is effetive in the tretment of osteorthritis n rheumtoi rthritis [24,43]. It is use for the tretment of erly stge hert filure, effetively reues the level of holesterol in the plsm, n inhiits the growth of rest ner [26,35]. Quine n Jpnese quine re plnts erive from the Fr Est. In Europe they hve een grown only reently; hene they re little ppreite y onsumers. Reserh in reent yers hs shown tht the ommon quine hs ntiteril n nti-llergeni properties, n n e use in the prevention n supportive tretment of illnesses suh s riovsulr isese, ronhil sthm n oughing [10,36,46]. It hs een use for long time in Chinese folk meiine for the tretment of epression, migrine n neurlgi. At the sme time, phrmologil stuies hve reporte the role of Jpnese quine in the prevention of gstri uler n triuspi neurlgi, n it hs shown promise in tretment of Prkinson s isese [15,48,49]. The present stuy reserhe in vitro iologil tivity of polyphenoli ethnol extrts of hokeerry, hwthorn, quine, Jpnese quine, lkurrnt n rosehip s soure of helthy sustnes. The im ws to investigte the stility of the extrts fter 12 months of storge t room temperture. Their ntioxint tivity ws exmine with respet to phosphtiylholine liposomes, simple moel of the iologil memrne, ntiril tivity in reltion to the free ril, n nti- -inflmmtory tivity in inhiition of enzymti oxition of linolei i, t the eginning n t the en of the storge perio. We lso evlute the ontent of polyphenols, flvonois n nthoynins in the extrts. Mterils n methos Trolox (6-hyroxy-2,5,7,8-tetrmethylhromne-2-roxyli i), iuprofen, (1,1-iphenyl-2-pirylhyrzyl), linolei i, lipoxise from Glyine mx (soyen), Folin-Ciolteu s phenol regent, glli i, rutin hyrte, Tween 20, trihloroeti i (TCA), n 2-thiorituri i (TBA) were purhse from Sigm-Alrih (Poznń, Poln). Egg yolk phosphtiylholine () ws otine from Lipi Prouts, UK. Aluminum hlorie hexhyrte (AlCl 3 6H 2 O) n tris(hyroxymethyl)-minomethne (TRIS) were otine from Chempur (Piekry Śląskie, Poln). Bori i n soium tetrorte were purhse from Crl Roth Krlsruhe, Germny. Preprtion of extrts The rw mterils for the stuy were hokeerry (Aroni melnorp (Mihx.) Elliott) fruit of the Gliin vriety, lkurrnt (Ries nigrum L.) of the Tisel vriety, hwthorn (Crtegus monogyn Jq.), rosehip (Ros nin L.), quine (Cyoni olong Mill.), n Jpnese quine (Chenomeles speios (Sweet) Nki). Fruits of hwthorn n rosehip were ollete in Szzytniki Gren in Wrołw. The fruits of quine n Jpnese quine were ollete in the Aroretum n Institute of Physiogrphy in Bolestrszye n in the Botnil Gren of Wrołw, respetively. Blkurrnt ws from the Luin re n hokeerry from the Sy Trzenikie plnttion. The fruits were frozen n then freeze-rie (ChRISTALPhA 1-4 LSC), n just efore extrtion they were isintegrte with n nlytil mill (A11 si of IKA-Werke, Germny). 1029

3 Postepy Hig Me Dosw (online), 2014; tom 68: Fruit extrts were otine in the following wy: 50 g of fruit lyophilizte ws ll overe in 200 ml of 70% wter-ethnol solution, sonite (20 khz, Soni, Itli) for 15 min, n the loholi extrt rine. The extrtion proess y sonition ws repete 2 more times. The extrt thus otine ws spun for 15 min in entrifuge (2500 rev/min) t room temperture n then the ethnol ws evporte to ry weight with rotry evportor (IKA RV 05B, Germny) for pprox. 50 min. Otine extrts of lkurrnt n hokeerry were issolve in istille wter n psse through olumn (70 m x 7.5 m) fille with Amerlite resin (XAD4). The olumn ws wshe with istille wter (out 2600 ml) until the wsh-out of totl sugrs (the onentrtion of sugrs in the extrtion, mesure with Poket PAL-1 refrtometer, me to zero). Anthoynin olornt ws otine fter wshing the olumn with 70% ethnol (out 1300 ml). The ollete frtion ws evporte in vuum evportor for out n hour until ry mss. The extrts thus otine were store t room temperture without light. Liposome oxition ssy The proeure ws presente in the pper y Grielsk et l. [12]. Lipi peroxition ws mesure s the thiorituri i retive sustne (TBARS) level, se on the metho of Buege et l. [6]. TBARS onentrtions were estimte using molr extintion oeffiient ε = 156 mm -1 m -1. The perentge of phosphtiylholine () liposome oxition inhiition ws lulte using the formul: % h = 100% where: A 0 is the onentrtion of mlonilehye (MDA) in smple without extrt e (ontrol) n A is the onentrtion of MDA in smple with extrt e, mesure t λ=535 nm. All mesurements were performe for two inepenent preprtions (n=4) using Cry 300 spetrophotometer (Vrin). The prmeter IC 50 ws etermine on the sis of plot showing the reltion etween the perentge of inhiition of lipi oxition n onentrtion of plnt extrts. This prmeter speifies the ntioxint onentrtion tht results in 50% inhiition of lipi oxition. Free-ril svenging ssy The effet of the stuie extrts on reution of ril onentrtion ws mesure spetrophotometrilly, s previously esrie y Brn-Willims et l. [5]. Briefly, methnol solution with sorption of pprox. 0.9 ws mixe with n pproprite mount of extrt, or Trolox solution, n immeitely ple in spetrophotometer. As ontrol, the sorption of (without ition of extrt) ws mesure t time t=0. Reution of in the smple fter 15 min inution with n ntioxint (of fixe onentrtion) ws etermine using the formul: % = 100% where: A 0 is the hnge of sorne t λ=517 nm, fter 15 min in the sene of n ntioxint; n A is the hnge in sorne t λ=517 nm, fter 15 min in the presene of n ntioxint. All etermintions were performe in six replites (n=6). Soyen lipoxygense inhiition Fruit extrts were teste on inhiition of soyen lipoxygense (1-LOX, Sigm, Poznń) tivity on the sis of the proeure esrie y Axelro et l. [1], with moifition. Fruit extrts t onentrtion of 8 µg ml -1 were use in the experiments. For omprtive purposes the non-steroil nti-inflmmtory gent iuprofen (use t 2.0 µg ml -1 ) ws lso teste. Retions were rrie out in 10 mm pth-length qurtz uvettes ontining, in finl volume of 2.6 ml: orte uffer ph 9.0, 0.1 mg ml -1 1-LOX, the extrts teste, n 50 μm linolei i (previously prepre in orte uffer ph 9.0 plus 1% Tween 20 n 0.1 M NOH). The inution ws rrie out for 3 min t room temperture. Referene uvettes (of 2.6 ml volume) ontine soium linolete orte uffer n n pproprite volume of solvent extrt teste (sustrte solution). Inhiition of 1-LOX tivity ws ssesse vi spetrophotometri monitoring of sorne inrese t 234 nm (t 3 min fter linolei i ition) ue to formtion of onjugte iene hyroperoxies (CDH) uring enzymti oxition proesses oring to Suin et l. [37]. All results re presente s the men ± SD of t lest two sets of inepenent experiments (n=6). Perentge inhiition ws lulte using the following formul: % h = 100% where: ΔA ontrol n ΔA smple enote the inrese of sorne 3 min fter sustrte ition to the proe without or with extrt teste, respetively. All results re presente s the men ± SD of t lest two sets of inepenent experiments (n=6). Totl phenols, flvonois n nthoynins ontent Totl polyphenols were etermine y the Folin-Ciolteu metho oring to Go et l. [13]. The results were lulte s mg of glli i equivlent in 1 g ry mss of extrt (mg GAE/g.m.). Mesurements were me using Cry 300 UV-VIS spetrophotometer (Vrin). Eh extrt ws nlyze for totl flvonoi ontent oring to the previously reporte olorimetri metho with moifitions y Lmison et l. [23]. The sorne of the solution ws mesure t 430 nm. The results were expresse s the rutin equivlent (mg RE/g.m.) n ompre with 1030

4 Strugł P., Grielsk J. Biologil tivity n stility in vitro... the rutin stnr urve, whih ws me uner the sme onitions. Totl nthoynin mount, lulte s yniin-3-o-gluosie, ws etermine y mens of the ph ifferentil metho of Trppey et l. [38]. Asorption ws mesure t 510 n 700 nm, using the formul: where: A = (A 510 -A 700 )ph 1.0 (A A 700 )ph 4.5. MW moleulr weight, DF ilution ftor, ε molr sorne. Results were expresse s mg equivlent of yniin-3-o- -gluosie in 1 g ry mss of extrt (mg C3GE/g.m.). Sttistil nlysis All results re expresse s men ± SD. The orreltion oeffiient etween totl phenolis ontent n ntioxint tivity ws lulte, n vrine nlysis ws one with the Dunn test. P vlues < 0.05 were onsiere sttistilly signifint. The progrm Sttisti 10.0 ws use for ll of the sttistil lultions. Results The work presents the results of stuy of polyphenoli extrts from selete fruits. The following hve een hrterize: the ntioxint properties of the extrts in the proess of uto-oxition of phosphtiylholine memrnes, the ntiril properties with respet to the free ril, n the nti-inflmmtory effet in enzymti oxition of linolei i. We lso etermine the totl ontent of polyphenols, flvonois, n nthoynins (in the se of hokeerry n lkurrnt). Antioxint tivities of extrts from the fruit of the hokeerry, hwthorn, rosehip, quine, Jpnese quine, n IC 50 [µg/ml] h = e e Chokeerry Hwthorn Quine Jpnese quine Blkurrnt egining of storge 12 month of storge Fig. 1. The ntioxint tivity (IC 50 ) of extrts from fruits of hokeerry, hwthorn, quine, Jpnese quine, lkurrnt n rosehip in the proess of utoxition liposomes for the initil time n fter 12 months of storge t room temperture. IC 50 vlues re the verges of two inepenent mesurements ± SD (n= 4). Different letters (-e) in the sme row inite signifint ifferenes (p < 0.05) Rose lkurrnt were expresse s IC 50. This prmeter is liste in Figure 1 for the strt of the storge perio, n fter 12 months. A low IC 50 vlue mens high ntioxint tivity of the extrt. The IC 50 prmeters for the strt of the storge perio of the extrts follow the sequene [µg ml -1 ]: Jpnese quine (19.38 ± 0.43) < hokeerry (27.69 ± 0.92) < lkurrnt (32.65 ± 0.16) < rosehip (0.68 ± 39.46) < hwthorn (47.05 ± 2.42) quine (45.71 ± 1.03, p < 0.05). Signifintly higher ntioxint tivity ws foun t the strt of storge for Jpnese quine n hokeerry extrts ompre to the tivity of hwthorn n quine (p<0.05). After 12-month storge the ntioxint tivity followe the sequene [µg ml -1 ]: lkurrnt (32.15 ± 0.25) > hokeerry (34.68 ± 0.14) Jpnese quine (34.89 ± 0.16) > quine (39.93 ± 0.67) > rosehip (41.32 ± 0.90) > hwthorn (49.36 ± 1.57) (p<0.05). Thus, there re ifferenes in tivity etween the strt n en of storge. Blkurrnt, hwthorn, rosehip, n quine prove to e stle extrts (ifferenes in the prmeter vrie from 1.5% to 12.6%). A lrge erese in tivity ws foun for Jpnese quine n hokeerry: 80 n 25%, respetively. Comprison of the ntioxint tivity of the extrts ws me in reltion to Trolox (IC 50 = 3.27 µg ml -1 ) s the positive ontrol (Tle 1) with the TEAA prmeter, expressing it in mm Trolox per g ry weight of rw mteril [mm g -1 ]. The higher the vlue of the prmeter the stronger the ntioxint property of n extrt. One n tell y these vlues tht ll extrts showe lower ntioxint tivity thn Trolox, from 6-fol (Jpnese quine) to 15-fol lower (hwthorn) t the eginning of the storge perio, n from 10 to 24 times lower fter 12 months of storge. The totl ontent of polyphenols in the extrts t the eginning of storge rnge from to [mg GAE/g.m.] for hwthorn n Jpnese quine, respetively. After 12 months of storge the ontents of polyphenols h not hnge shrply, with the exeption of Jpnese quine (pprox. 53% eline). Anlysis of the results otine onfirme the existene of strong orreltion etween ntioxint tivity (IC 50 ) n the totl polyphenols ontent t the eginning of storge of the plnt extrts (r = ) n reltively strong orreltion fter 12 months (r = ), t signifine level p < The ontent of flvonois in the extrts showe tht the rihest re extrts of hokeerry ( ± 0.31 mgre/g. m) n lkurrnt (79.03 ± 0.19 mgre/g.m.). The remining 4 extrts ontine muh smller mounts of flvonois; they rnge from 9.10 ± 0.02 to 4.71 ± 0.19 mgre/g.m. For extrts of hokeerry n lkurrnt the nthoynins ontent ws ssye to e 1.29 ± 0.01 to 0.92 ± 0.03 mg C3GE/g.m. The results on ntiril tivity of the extrts re shown in Figure 2, where the vlues of the prmeter IC 50 t the eginning of the storge perio n fter 12 months re ompile. It shoul e note tht of ll the stuie plnt extrts the highest tivity to eliminte the free ril ws shown y: Jpnese quine, rosehip, hokeerry n quine. In ft, there ws sli- 1031

5 Postepy Hig Me Dosw (online), 2014; tom 68: Tle 1. The ntioxint n ntiril tivity of stuie fruits extrts expresse s TEAA [mm TE/g.m.] n TEAC [mm TE/g.m.], respetively. Plnts TEAA [mm TE/g.m.] TEAC [mm TE/g.m.] egining of storge 12 month of storge egining of storge 12 month of storge Chokeerry 0.47 ± ± ± ± 0.02 Hwthorn 0.27 ± ± ± ± 0.13 Quine 0.29 ± ± ± ± 0.05 Jpnese quine 0.67 ± ± ± ± 0.01 Blkurrnt 0.40 ± ± ± ± 0.03 Rosehip 0.33 ± ± ± ± 0.06 ght hnge in the IC 50 prmeter with the exeption of hwthorn extrt, for whih its vlue signifintly erese y 33% fter 12 months of storge. The results of ntiril tivity t the eginning of storge n fter 12 months, expresse in Trolox equivlent s the TEAC prmeter (Tle 1), inite weker tivity of ll the extrts exmine in omprison with Trolox (IC 50 = 4.87 µg ml -1 ). It shoul e note tht the results of ntiril tivity orrelte well with the totl polyphenol ontent in the test extrts. Correltion oeffiients re r = t the eginning of storge n r = fter 12 months (p < 0.05). Inhiition of LOX [%] Chokeerry Hwthorn Quine Jpnese quine Blkurrnt Rose 16 egining of storge 12 month of storge IC 50 [µg/ml] Fig. 3. The perentge of inhiition of 1-LOX tivity (nti-inflmmtory tivity), use y extrts of hokeerry, hwthorn, quine, Jpnese quine, lkurrnt n rosehip t 8 µg ml -1 onentrtion. The perentge of inhiition re given s verge of two inepenent mesurements ±SD (n= 6) Different letters (-) in the sme row inite signifint ifferenes (p < 0.05) 2 0 Chokeerry Hwthorn Quine Jpnese quine Blkurrnt egining of storge 12 month of storge Rose uses 56% inhiition t onentrtion of 4-fol less thn the test extrts (2 µg ml -1 ). Fig. 2. Comprison of IC 50 prmeters in the neutrliztion proess of the free rils use y extrts of hokeerry, hwthorn, quine, Jpnese quine, lkurrnt n rosehip. The vlues re given s the verges of two inepenent mesurements ± SD (n= 6). Different letters (-) in the sme row inite signifint ifferenes (p< 0.05) Figure 3 shows the results of nti-inflmmtory tivity of the plnt extrts when pplie t onentrtion of 8 µg ml -1. The highest tivity of 1-LOX inhiition t the eginning of the storge perio n fter 12 months showe rosehip extrts (up to 46% n 33% inhiition, respetively) n hwthorn (respetively 39 n 31% inhiition). Inhiition of the extrts fter 12 months of storge roppe from 6.3% to 77.3%, with the highest eline in Jpnese quine tivity. Comprison of the inhiition of extrts in reltion to iuprofen s positive ontrol showe tht it Disussion The use of sustnes erive from plnts in prevention n tretment of helth prolems shoul e preee y etile stuies. Therefore, polyphenoli ompouns extrte from the fruit of lkurrnt, hokeerry, hwthorn, rosehip, Jpnese quine n quine were teste in vitro uring 12 months of storge t room temperture. The extrts were teste for their ntioxint pity in reltion to phospholipi moel memrnes n lso for inhiition of soyen lipoxygense enzyme tivity, whih is goo initor of inflmmtory retions in the oy. We lso exmine the ility of the extrts to eliminte the free ril, whih showe the extrts ntioxint tion s svengers of free rils in the reox proess. This stuy of ntioxint tivity in reltion to the proess of utooxition of phosphtiylholine liposomes showe 1032

6 Strugł P., Grielsk J. Biologil tivity n stility in vitro... signifintly higher tivity of lkurrnt, hokeerry n Jpnese quine extrts in omprison to the remining quine, rosehip n hwthorn extrts, fter 12 months of storge. It shoul e e tht lthough extrts of hokeerry n Jpnese quine showe the lrgest egrtion fter tht perio of storge, they still possesse reltively high ntioxitive potentil. The effetiveness of polyphenoli extrts s ntioxints in the proess of oxition of lipi strutures hs een the sujet of reltively numerous stuies, inluing with the use of moel lipi memrnes [11], tht onfirme the high tivity of polyphenoli ompouns s svengers of free rils. It hs een suggeste tht the hyrophili omponents of the extrts lolize on the surfe of the lipi ilyer, neutrlizing free rils tht rrive there from the externl environment [41]. Other more hyrophoi omponents of the extrts re ple of penetrting eep into the ilyer [34] n regenerting lipi oxies n peroxies tht rise in propgtion of the oxition proess. The use of nturl ntioxints to inhiit lipi peroxition hs unquestionle vntges over syntheti ntioxints even if the former shoul e use in muh higher onentrtions ompre to the onentrtions of syntheti ntioxints suh s Trolox or BHT. Stuies on pplition of BHT hve in ft shown toxi tion of this syntheti ntioxint [18,22]. The extrts, eing mixture of severl ingreients inluing the flvonols frtion, n to vrying egree penetrte the lipi memrne n ffet repir of its interior. Therefore, the results of reserh of ntioxint tivity in reltion to moel liposome memrnes, whih re simple moel of the ell memrne, n e helpful in preiting the enefiil tion of these sustnes in vivo. The results of the ntiril tivity of the extrts inite tht the highest ility to svenge the ril is shown y extrts of Jpnese quine n rosehip. The high orreltion oeffiients etween totl polyphenol ontent n test results, t the eginning of the storge perio (r = ) n fter 12 months (r = ), show the highest tivity for extrts whih ontine the most polyphenoli ompouns, whih is in line with the reports of other uthors [9]. The helth-supporting tivity of ompouns of the polyphenol group is ue to their ility to inhiit the 1-LOX enzyme tht tlyses oxition of ftty is. Retion of this enzyme with linoleni i proues sustnes tht prtiipte in the formtion of mny iseses inluing rterioslerosis, inflmmtory owel isese, psorisis n sthm [30]. Inhiition of 1-LOX enzyme tivity my therefore inite the potentilly enefiil effets of nturl polyphenoli ompouns, inluing t the level of the oy. The highest tivity to inhiit 1-LOX ws oserve for the rosehip extrt (46% inhiition) n hwthorn (38% inhiition). Rosehip extrt showe out 10% lower tivity thn iuprofen (pprox. 56%) s positive ontrol t 2 µg ml -1 onentrtion. It is reporte y vrious uthors tht rosehip is rih in queretin erivtives, kempferol s well s glli i n ellgi i [39,40]. The presene of these sustnes in rosehip extrt is lso onfirme y our reserh (t not pulishe). Anti-inflmmtory effets of queretin n glli i hve lso een onfirme y inepenent reserhers [7,32]. The nti-inflmmtory effet of rosehip extrt is onfirme y in vivo stuies performe on rts. They showe tht ethnol-wter extrt from tht fruit inhiits the evelopment of rrgeenn-inue eem in rts [25]. The min sustnes ontine in fruits of hwthorn re queretin-3-o-gltosie, queretin-3-o- -gluosie n hlorogeni i [27,28]. It is elieve tht the nti-inflmmtory properties of hwthorn extrt re lrgely ue to one of the omponents, i.e. queretin-3-o- -gltosie. Its nti-inflmmtory properties in vivo, in n experiment on inflmmtion inue in the liver of rts, hs een onfirme in the literture [19]. 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