Phytochemical and pharmacological study of Sedum dendroideum leaf juice

Phytochemical and pharmacological study of Sedum dendroideum leaf juice
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  Journal of Ethnopharmacology 102 (2005) 217–220 Phytochemical and pharmacological study of  Sedum dendroideum  leaf juice Giany O. De Melo a , David do C. Malvar b , Frederico A. Vanderlinde b , Priscilla A. Pires b ,Wellington S. Cˆortes b , Pedro Germano Filho c , Michelle F. Muzitano a ,Carlos R. Kaiser d , Sˆonia S. Costa a , ∗ a  N´ ucleo de Pesquisas de Produtos Naturais, Universidade Federal do Rio de Janeiro, 21 941-590 Rio de Janeiro, RJ, Brazil b  Departamento de Ciˆ encias Fisiol´ ogicas, Instituto de Biologia, Universidade Federal Rural do Rio de Janeiro, Serop´ edica 23 850-000, RJ, Brazil c  Departamento de Botˆ anica, Instituto de Biologia, Universidade Federal Rural do Rio de Janeiro, Serop´ edica 23 850-000, RJ, Brazil d  Instituto de Qu´ımica, Universidade Federal do Rio de Janeiro, CP 068534, 21 941-972 Rio de Janeiro, RJ, Brazil Received 3 September 2004; received in revised form 25 May 2005; accepted 9 June 2005Available online 28 July 2005 Abstract The fresh juice from leaves of   Sedum dendroideum  Moc & Sess´e (Crassulaceae) is used in Brazilian traditional medicine for the treatmentof gastric and inflammatory disorders. The present investigation was carried out to evaluate in vivo antinociceptive and anti-inflammatoryactivities of this plant material. The oral administration (0.1–1g/kg) of the lyophilized  Sedum dendroideum  juice (LJ) caused a significantdose-related reduction of acetic acid-induced writhing response (ID 50  =631mg/kg) and inhibited croton oil-induced ear oedema formation(66% inhibition at 1g/kg) in mice. In the formalin-induced nociception in mice, LJ (1g/kg) only inhibited the second phase of nociception(46%). Phytochemical investigation revealed four known kaempferol glycosides, here, described at the first time for this species. Theseflavonoids probably explain the antinociceptive and anti-inflammatory effects of the fresh juice of   Sedum dendroideum .© 2005 Elsevier Ireland Ltd. All rights reserved. Keywords: Sedum dendroideum ; Crassulaceae; Kaempferol glycosides; Antinociceptive activity; Anti-inflammatory activity 1. Introduction Fresh juice of leaves from  Sedum dendroideum  Moc &Sess´e (syn.:  Sedum praealtum  DC.) is popularly used to treatulcer (Carlini et al., 1970), general inflammatory troubles(Camargo et al., 2002) and as a contraceptive (Silva-Torres et al., 2003). Sedum  species (Crassulaceae) have been used in popu-lar medicine against different disease symptoms (Niemannet al., 1976). They are known to contain alkaloids, tannins,flavonoidsandcyanogeniccompounds(Niemannetal.,1976;Nahrstedt et al., 1982; Mulinacci et al., 1995; Stevens et al.,1995). Among all the  Sedum  species that have been investi-gated,  Sedum dendroideum  has not received great attention. ∗ Corresponding author. Tel.: +55 21 2562 6512; fax: +55 21 2562 6512.  E-mail address: (S.S. Costa). Considering the ethnopharmacological profile of thisgenus, the aim of this work was to assess the antinociceptiveand anti-inflammatory activities and the chemical composi-tion of that species. Here we report our first results of chem-ical and pharmacological investigations undertaken with afresh juice from leaves of   Sedum dendroideum . 2. Materials and methods 2.1. Chemical proceduresSedum dendroideum  leaves, out of the flowering season,were collected at S˜ao Paulo, SP (Brazil) in December 2001.A voucher specimen (RBR no. 7401) was deposited in theherbarium of the Department of Botany (UFRRJ, Brazil). 0378-8741/$ – see front matter © 2005 Elsevier Ireland Ltd. All rights reserved.doi:10.1016/j.jep.2005.06.015  218  G.O. De Melo et al. / Journal of Ethnopharmacology 102 (2005) 217–220 Fresh leaves (2.12kg) were expressed in a food pro-cessor. The juice was separated by filtration until a trans-parent yellow liquid was obtained. This juice (1.9l) wasfrozen and lyophilized (LJ). LJ (44.9g) was separatedon an RP-2 column (9.5cm × 3.8cm), affording two frac-tions: FW (2.8l; eluted with water) and FM (1.8l; elutedwith H 2 O/MeOH gradient until 100% methanol). FM(1.26g) was purified on an RP-2 column (22.5cm × 2.4cm)affording seven fractions. The FM2, FM3 and FM4 frac-tions showed flavonoid spots under thin layer chromato-graphy.Fraction FM2, eluted with H 2 O/MeOH 8:1, was purifiedon an RP-2 column (45cm × 1.2cm) affording four frac-tions. The second fraction (H 2 O/MeOH 9:1; 101.7mg) waschromatographed on an RP-18 column (36cm × 1cm). Thesecondfraction,elutedwithH 2 O/MeOH7:3,waspurifiedonaSephadexLH-20(20.5cm × 1cm;H 2 O/MeOH5:5)giving7.9mg of pure flavonoid [ 1 ].Fraction FM3, eluted with H 2 O/MeOH 9:1, was puri-fied on an RP-2 column (18cm × 1.5cm) affording fivefractions. The third fraction (H 2 O/MeOH 9:1) gave aflavonoid, which was purified on a Sephadex LH-20 column(25.7cm × 0.9cm;H 2 O/MeOH3:7)yieldingthreefractions.The second fraction gave 8.2mg of flavonoid [ 2 ].From FM4, eluted with H 2 O/MeOH 7:3, was obtained ayellow crystal that was separated by centrifugation, washedwith distilled water and lyophilized. The pellet afforded249mg of flavonoid [ 3 ].The FM4 mother-liquors, after concentration in a water-bath, gave flavonoid [ 4 ] as yellow crystals (27mg) thatwere separated by centrifugation and washed with distilledwater.These flavonoids were identified by comparison of their 1 H and  13 C NMR data with the literature (Nakano et al.,1981;Agrawal,1989;Mulinaccietal.,1995;Pizzolattietal.,2003).Chromatography was performed on RP-2 (70–230 mesh,Merck) or RP-18 (40–60  M, Merck) silanized silica(using H 2 O/MeOH gradient) and on Sephadex LH-20 gel(25–100nm, Sigma).  1 H and  13 C NMR spectra (DMSO- d  6 signals as internal reference:  δ  2.50 for  1 H NMR and  δ  40.0for  13 C NMR) were recorded on a Bruker DRX-300 ( 1 H:300MHz;  13 C: 75MHz) or on a Bruker Avance DRX-400( 1 H: 400MHz;  13 C: 100MHz) spectrometer. 2.2. Pharmacological procedures2.2.1. Animals Adult male Swiss mice (25–35g) were housed in plasticcages, with food and tap water available ad libitum, in thecolony room. Mice were acclimatized to the laboratory forat least 60min prior to the test procedure and were fasted for12–18h before the gavage. All experiments were carried outin accordance with current guidelines for the care of labora-tory animals and the ethical guidelines on the use of animalsin pain research (Zimmermann, 1983). 2.2.2. Drugs Formalin, acetic acid, acetone (Merck AG, Darmstadt,Germany), croton oil, indomethacin (Sigma Chemical Co.,St. Louis, MO, USA), fentanyl (Janssen Pharmaceutical),diazepam (Crist´alia, SP, Brazil), LJ and drugs were dilutedin water or saline. 2.2.3. Acetic acid-induced writhing test  Group of six mice were treated orally with water(10ml/kg), LJ (1g/kg) or indomethacin (10mg/kg) 60minbefore acetic acid injection (1.2%, 0.1ml/10g) and the num-ber of writhes was counted for the following 30min (Kosteret al., 1959). 2.2.4. Hot-plate test  The latency (s) of heat stimulus was measured every30min, starting 30min before and up to 2h after pre-treatment with LJ (1g/kg, p.o.), fentanyl (100  g/kg, s.c.)or water (10ml/kg, p.o.) (D’Amour and Smith, 1941). 2.2.5. Formalin-induced nociception Groups of 6–10 mice were orally treated with water, LJ(1g/kg) or indomethacin (10mg/kg), 60min prior to injec-tion of the formalin solution (3%; 20  l/paw; s.c.) into theplantar surface of the hind paw ( injection). The timethat animals spent licking the injected paw was measuredwith a chronometer and was considered as an index of pain.From the formalin injection, the initial nociceptive responsepeaked at about 5min (first phase) and was followed bya second peak (second phase) that occurred at 15–30minpost injection (Hunskaar et al., 1986; Hunskaar and Hole,1987). 2.2.6. Edema induced by croton oil in mouse ear  One hour after oral administration of water, LJ (1g/kg)or indomethacin (10mg/kg), each animal ( n =6) was treatedwith10  loffreshlypreparedcrotonoil(2.5%inacetone)ontheinnersurfaceoftherightear.Theleftearwastreatedwiththe same volume of acetone (control) (Tubaro et al., 1986). 2.2.7. Rota-rod test  Mice( n =10)weretreatedwithvehicle(water),LJ(1g/kg,p.o.)ordiazepam(3mg/kg,s.c.).Rota-rodtestwasemployedaccording to the previously described procedure (Rosland etal., 1990). 2.2.8. Statistical analysis of data Results were expressed as means ± S.E.M. The ID 50  val-ueswerepresentedasgeometricmeansandconfidencelimits.Differences between two means were determined using theStudent’s  t  -test. Differences between more than two meanswere calculated using one-way analysis of variance followedby the Tukey–Kramer test, and test data were considered dif-ferent at a significance level of   p <0.05 (Sokal and Rohlf,1981).  G.O. De Melo et al. / Journal of Ethnopharmacology 102 (2005) 217–220  219 3. Results and discussion 3.1. Phytochemical investigation Four flavonoids were obtained (Fig. 1): kaempferol 3- O -  -rhamnopyranosyl-(1  →  2)-  -glucopyranoside-7- O -  -glucopyranoside[ 1 ],kaempferol3- O -  -rhamnopyranosyl-(1 → 2)-  -glucopyranoside-7- O -  -rhamnopyranoside[ 2 ],kae-mpferol 3- O -  -rhamnopyranoside-7- O -  -rhamnopyrano-side (kaempferitrin) [ 3 ] and kaempferol 3- O -  -gluco-pyranoside-7- O -  -rhamnopyranoside [ 4 ]. Kaempferitrinwas the most abundant in this species (0.55% from LJ).Flavonoids have long been recognized to possess a widevariety of biological activities such as antioxidant and anti-inflammatory (Middleton et al., 2000). Hence, the pres-ence of kaempferitrin, known to have anti-inflammatory andantioxidant activities (Toker et al., 2004), principally, couldcontribute favourably to the medicinal use of   Sedum den-droideum . While  4  is reported for the first time in the Cras-sulaceae family, kaempferol glycosides  1 – 3  were previouslyreported from another  Sedum  species (Niemann et al., 1976;Mulinacci et al., 1995). 3.2. Pharmacological investigation LJ (0.1, 0.3 and 1g/kg) produced a dose-related inhibi-tion of acetic acid-induced abdominal writhing (10 ± 1.5,42.4 ± 4.3and54.5 ± 7.4%,respectively;IC 50  =631mg/kg),compared to the control group (44 ± 4.3 writhes). The grouptreated with indomethacin showed a decrease in accumu-lated abdominal writhes (15 ± 3.6). In the hot-plate test,the basal latency of the control group was 6.7 ± 0.4s andLJ (1g/kg, p.o.) did not alter the pain latency of the ani-mals even at a dose level high enough to induce a max- Fig. 1. Chemical structure of flavonoids isolated from  Sedum dendroideum .Rha, rhamnose and Glu, glucose. imal response with the acetic acid test. The positive con-trol fentanyl increased the latency of heat stimulus by 3.3-fold (30min) and 2.8-fold (60min). In the formalin-inducednociception test, LJ (1g/kg) pre-treatment did not modifythe reactivity during first phase (neurogenic pain) of noci-ceptive response (140.1 ± 7.3s). However, LJ significantlyinhibited 46.5% of the second phase (inflammatory pain) of nociception (132.3 ± 15.8s) compared to the control group(142.4 ± 15.2s and 247.4 ± 32.9s, first and second phasesrespectively). Indomethacin reduced 62.8% of the reactiv-ity during the second phase. To assess the antioedemato-genic effect of LJ, the croton oil-induced mouse ear oedemamethod was used. The difference between right and left earweight obtained in the control group was 7.3 ± 1.2mg, andLJ(1g/kg)orindomethacinpre-treatmentinhibitedtheoede-matogenic response after topical application of croton oilby 66% (2.5 ± 0.6mg) and 42.3% (3.1 ± 0.8mg), respec-tively. In the rota-rod test, control vehicle response was58.3 ± 1.7s versus 57.5 ± 1.5s in the presence of LJ (1g/kg,p.o.), showing that LJ did not cause any significant effect onthe performance of the animals. Under the same experimen-talconditions,diazepam(3mg/kg,s.c.)reducedsignificantly(  p <0.05) by 27.6 ± 3.9% the motor response of the animals(42.2 ± 6.1s).Few studies validating the folk medicinal use of   Sedumdendroideum  have been done. Studies on this speciesshowed spermicidal and anti-inflammatory activities fromaerial parts extracts (Camargo et al., 2002; Silva-Torreset al., 2003). However, Camargo et al. (2002) reported the anti-inflammatory activity for a hot aqueous extract,using different experimental models when compared withthe present one. In our study, oral treatment with  S. den-droidem  fresh juice, in accordance with popular use, inhib-ited croton oil oedema formation in mice, suggesting ananti-inflammatory activity. Furthermore, this juice also pro-duced potent antinociceptive activity against acetic acid-and formalin-induced pain response in mice, inhibiting theinflammatory components of the formalin test, without inter-fering with the hot-plate responses. 4. Conclusions Thisstudyshowedthatthejuicefrom Sedumdendroideum leaves shows anti-inflammatory and antinociceptive activ-ities. Both effects seem to be related to an inhibition of prostaglandin synthesis because the plant extract reduced theinflammatory responses induced by either croton oil or for-malin second phase, as well as pain stimuli induced by aceticacid. The four flavonoids here described may explain theantinociceptive and anti-inflammatory effects from  S. den-droideum . Our findings could explain its popular use againstpain and inflammatory diseases. A bioassay-guided fraction-ation of the crude juice is now in progress to identify thebioactive substance(s).  220  G.O. De Melo et al. / Journal of Ethnopharmacology 102 (2005) 217–220 Acknowledgements G.O. De Melo thanks CNPq (PIBIC) and CAPES (Ph.D.)for fellowships. References Agrawal, P.K., 1989. Carbon-13 NMR of Flavonoids. Elsevier SciencePublishers B.V., Amsterdam, The Netherlands.Camargo, M.E.M., Romero, M.B., Zamora, D.R., Carrillo, P.C., Maldon-ado, M.E.V., 2002. Study of the anti-inflammatory effect of   Sedum praealtum  (Siempreviva) in the rat: dose-dependent response. Pro-ceedings Western Pharmacology Society 45, 129–130.Carlini, E.A., Neto, J.P., Almeida, E.T., Marigo, C., 1970. ´Ulcera porcontenc¸˜ao em ratos: ac¸˜ao protetora de extrato aquoso de “b´alsamo”. Estudo preliminar. Anais da Academia Brasileira de Ciˆencias 42,267–270.D’Amour, F.E., Smith, D.L., 1941. A method for determining loss of painsensation. The Journal of Pharmacology and Experimental Therapeu-tics 72, 74–79.Hunskaar, S., Berger, O.G., Hole, K., 1986. Dissociation betweenantinociceptive and anti-inflammatory effects of acetylsalicylic acidand indomethacin in the formalin test. Pain 25, 125–132.Hunskaar, S., Hole, K., 1987. The formalin test in mice: dissociationbetween inflammatory and non-inflammatory pain. Pain 30, 103–114.Koster, R., Anderson, M., De Beer, E.J., 1959. Acetic acid for analgesicscreening. Federation Proceedings 18, 412.Middleton Jr., E., Kandaswami, C., Theoharides, T.C., 2000. The effectsof plant flavonoids on mammalian cells: implications for inflamma-tion, heart disease, and cancer. Pharmacological Reviews 52, 673–751.Mulinacci, N., Vincieri, F.F., Baldi, A., Bambagiotti-Alberti, M., Sendl,A., Wagner, H., 1995. Flavonol glycosides from  Sedum telephium subspecies  maximum  leaves. Phytochemistry 38, 531–533.Nahrstedt, A., Walther, A., Wray, V., 1982. Sarmentosin epoxide, anew cyanogenic compound from  Sedum cepae . Phytochemistry 21,107–110.Nakano, K., Murakami, K., Nohara, T., Tomimatsu, T., Kawasaki, T.,1981. The constituents of   Paris verticillata  M.v. Bieb. Chemical andPharmaceutical Bulletin 29, 1445–1451.Niemann, G.J., Visser-Simons, J.M.J., Hart, H., 1976. Flavonoids of somespecies of   Sedum . Planta Medica 30, 384–387.Pizzolatti, M.G., Cunha Jr., A., Szpoganicz, B., Souza, E., Braz-Filho,R., Schripsema, J., 2003. Flavon´oides glicosilados das folhas e flo-res de  Bauhinia forficata  (Leguminosae). Qu´ımica Nova 26, 466–469.Rosland, J.H., Hunskaar, S., Hole, K., 1990. Diazepam attenuates mor-phine antinociception test-dependently in mice. Pharmacology andToxicology 66, 382–386.Silva-Torres, R., Montellano-Rosales, H., Ramos-Zamora, D., Castro-Mussot, M.E., Cerda-Garcia-Rojas, C.M., 2003. Spermicidal activityof the crude ethanol extract of   Sedum praealtum  in mice. Journal of Ethnopharmacology 85, 15–17.Sokal, R.R., Rohlf, F.J., 1981. Biometry: The Principle and Practice of Statistics, second ed. W.H. Freeman, New York, p. 859.Stevens, J.F., Hart, H., Van Ham, R.C.H.J., Elema, E.T., Van Den Ent,M.M.V.X., Wildeboer, M., Zwaving, J.H., 1995. Distribution of alka-loids and tannins in the Crassulaceae. Biochemical Systematics andEcology 23, 157–165.Toker, G., K¨upeli, E., Memiso˘glu, M., Yesilada, E., 2004. Flavonoidswith antinociceptive and anti-inflammatory activities from the leavesof   Tilia argentea  (silver linden). Journal of Ethnopharmacology 95,393–397.Tubaro, A., Dri, P., Melato, M., Mulas, G., Bianchi, P., Del Negro, P.,Della Loggia, R., 1986. In the croton oil ear test the effects of nonsteroidal anti-inflammatory drug (NSAIDs) are dependent on the doseof the irritant. Agents Actions 19, 371–373.Zimmermann, M., 1983. Ethical guidelines for investigations of experi-mental pain in conscious animals. Pain 16, 109–110.
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