Antioxidant and Hepatoprotective Effects of Cyathea phalerata Mart. (Cyatheaceae)
Mariana Appel Hort, Silvia DalBó, Inês Maria Costa Brighente, Moacir Geraldo Pizzolatti, Rozangela Curi Pedrosa,
Rosa Maria Ribeiro-do-Valle,
Rozangela Curi Pedrosa
Department of Biochemistry, CCB, Federal University of Santa Catarina, Florianópolis, SC, Brazil
Search for more papers by this authorMariana Appel Hort, Silvia DalBó, Inês Maria Costa Brighente, Moacir Geraldo Pizzolatti, Rozangela Curi Pedrosa,
Rosa Maria Ribeiro-do-Valle,
Rozangela Curi Pedrosa
Department of Biochemistry, CCB, Federal University of Santa Catarina, Florianópolis, SC, Brazil
Search for more papers by this authorAuthor for correspondence: Rosa Maria Ribeiro-do-Valle, Department of Pharmacology, CCB, Block ‘D’, Federal University of Santa Catarina, Campus Universitário Trindade, Florianópolis, CEP 88.049-900, SC, Brazil (fax +5548 3337 5479, e-mail [email protected]).
Abstract
Abstract: The present study investigated the antioxidant properties of Cyathea phalerata Mart. (Cyatheaceae) using in vitro and in vivo assays. The in vitro antioxidant potential of the crude extract (CE), precipitate (PPT), aqueous fraction (AQF), n-butanolic fraction (BUF) and ethyl acetate fraction (EAF) from C. phalerata was evaluated through the scavenging of diphenyl-1-picryl-hydrazyl-hydrate (DPPH), superoxide anion (O2•−) (nitroblue tetrazolium assay) and hydroxyl radicals (OH•) (deoxyribose assay), and lipid peroxidation in rat liver homogenate. In these assays, it was observed that EAF had marked antioxidant potential, especially as a scavenger of the OH• radical and in inhibiting lipid peroxidation. The in vivo evaluation of oxidative stress (DNA fragmentation, membrane lipoperoxidation and carbonyl protein formation) and the antioxidant defenses (concentration of reduced glutathione, as well as catalase and glutathione S-transferase activities) were measured in mice pre-treated with EAF (10, 30 or 100 mg/kg, orally) and later exposed to carbon tetrachloride (CCl4). The EAF decreased thiobarbituric acid reactive substances levels, DNA damage and carbonyl protein contents, and increased catalase and glutathione S-transferase activities. Based on these results, it is concluded that the EAF from C. phalerata protects liver from oxidative stress induced by CCl4 in mice and these effects are probably related to the antioxidant activity associated with the free radical scavenging property of this fraction.
References
- 1 Arai Y, Koide N, Ohki F, Ageta H, Yang LL, Yen KY. Ferns constituents: triterpenoids isolated from leaflets of Cyathea spinulosa. Chem Pharm Bull 1994; 42: 228–32.
- 2 Arai Y, Hirohara M, Mtsuhira M, Tyoasaki K, Ageta H. Ferns constituents: triterpenoids isolated from leaflets of Cyathea lepifera. Chem Pharm Bull 1995; 43: 1849–52.
- 3 Hiraoka A, Hasegawa M. Flavonoid glycosides from five Cyathea species. Bot Mag Tokio 1975; 88: 127–30.
- 4 Hiraoka A, Maeda M. A new acylated flavonol glycoside from Cyathea contaminans Copel. and its distribution in the Pterophyta. Chem Pharm Bull 1979; 27: 3130–6.
- 5 Bringmann G, Günther C, Jumbam DN. Isolation of 4-O-beta-D-glucopyranosylcaffeic acid and gallic acid from Cyathea dregei Kunze (Cyatheaceae). Pharm Pharmacol Lett 1999; 9: 41–3.
- 6 Pizzolatti MG, Brighente, IMC, Bortoluzzi AJ, Schripsema J, Verdi LG. Cyathenosin A, a spiropyranosil derivate of protocatechuic acid from Cyathea phalerata. Phytochemistry 2007; 68: 1327–30.
- 7 Cazarolli LH, Zanatta L, Jorge AP, De Sousa E, Horst H, Woehl VM et al . Follow-up studies on glycosylated flavonoids and their complexes with vanadium: their anti-hyperglycemic potential role in diabetes. Chem Biol Interact 2006; 163: 177–91.
- 8 Middleton EJR, Kandaswami C, Theoharides, TC. The effects of plant flavonoids on mammalian cells: implications for inflammation, heart disease, and cancer. Pharmacol Rev 2000; 52: 673–751.
- 9 Williams RJ, Spencer JPE, Rice-Evans C. Flavonoids: antioxidants or signalling molecules? Free Radic Biol Med 2004; 36: 838–49.
- 10 Soobrattee MA, Neergheen VS, Luximon-Ramma A, Aruoma OI, Bahorun T. Phenolics as potential antioxidant therapeutic agents: mechanism and actions. Mutat Res 2005; 579: 200–13.
- 11 Halliwell B, Gutteridge JMC. Free Radicals in Biology and Medicine. Oxford University Press, Oxford, UK, 1999.
- 12 Valko M, Leibfritz D, Moncol J, Cronin MTD, Mazur M, Telser J. Free Radicals and antioxidants in normal physiological functions and human diseases. Int J Biochem Cell Biol 2007; 39: 44–84.
- 13 Pietta P. Flavonoids as antioxidants. J Nat Prod 2000; 63: 1035–42.
- 14
Nes WD,
Norton RA,
Benson M.
Carbon-13 NMR studies on sitototerol biosynthesized from mevalonates.
Phytochemistry
1972; 31: 805–11.
10.1016/0031-9422(92)80019-B Google Scholar
- 15 Agrawal KP. Carbon-13 of Flavonoids: Studies in Organic Chemistry 39. Elsevier Science, New York, 1989.
- 16 Agrawal KP. NMR spectroscopy in the structural elucidation of oligosaccharides and glycosides. Phytochemistry 1992; 31: 3307–30.
- 17 Mensor LL, Menezes FS, Leitao GG, Reis AS, Dos Santos TC, Coube CS et al . Screening of Brazilian plant extracts for antioxidant activity by the use of DPPH free radical method. Phytother Res 2001; 15: 127–30.
- 18 Robak J, Gryglewski RJ. Flavonoids are scavengers of superoxide anions. Biochem Pharmacol 1988; 37: 837–41.
- 19 Halliwell B, Gutteridge JMC. Formation of a thiobarbituric-acid-reactive substance from deoxyribose assay in the presence of iron salts – the role of superoxide and hydroxyl radicals. FEBS Lett 1981; 128: 347–52.
- 20 Chen H, Tappel AL. Protection of multiple antioxidants against heme protein oxidation and lipid peroxidation induced by CBrCl3 in liver, lung, heart and spleen. J Agric Food Chem 1996; 44: 854–8.
- 21 Yoshitake I, Ohishi E, Sano J, Mori T, Kubo K. Effects of KF-14363 on liver fibrosis in rats with chronic liver injury induced by carbon tetrachloride. J Pharmacobiodyn 1991; 14: 679–85.
- 22 Hermes-Lima M, Willmore WG, Storey KB. Quantification of lipid peroxidation in tissue extracts based on Fe(III)xylenol orange complex formation. Free Radic Biol Med 1995; 19: 271–80.
- 23 Singh NP, Mccoy MT, Tice RR, Schneider El. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 1988; 175: 184–91.
- 24 Nadin SB, Vargas-Roig LM, Ciocca DR. A silver staining method for single-cell gel assay. J Histochem Cytochem 2001; 49: 1186–93.
- 25 Reznick AZ, Packer L. Carbonyl assays for determination of oxidatively modified proteins. Methods Enzymol 1994; 233: 357–63.
- 26 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193: 265–75.
- 27 Anderson ME. Determination of glutathione and glutathione disulfide in biological samples. Methods Enzymol 1985; 113: 548–55.
- 28 Habig WH, Pabst MJ, Jakoby WB. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem 1974; 249: 7130–9.
- 29 Aebi H. Catalase in vitro. Methods Enzymol 1984; 105: 121–6.
- 30 Halliwell, B. The antioxidant paradox. Lancet 2000; 355: 1179–80.
- 31 Halliwell B, Rafter J, Jenner A. Health promotion by flavonoids, tocopherols, tocotrienols, and other phenols: direct or indirect effects? Antioxidant or not? Am J Clin Nutr 2005; 81: 268–76.
- 32 Nijveldt RJ, Van Nood E, Van Hoorn DE, Boelens PG, Van Norren K, Van Leeuwen, PA. Flavonoids: a review of probable mechanisms of action and potential applications. Am J Clin Nutr 2001; 74: 418–25.
- 33 Butkovic V, Klasinc L, Bors W. Kinetic study of flavonoid reactions with stable radicals. J Agric Food Chem 2004; 52: 2816–20.
- 34 Nardi GM, Felippi R, DalBó S, Siqueira-Júnior JM, Arruda DC, Delle Monache F et al . Anti-inflammatory and antioxidant effects of Croton ceutidifolius bark. Phytomedicine 2003; 10: 176–84.
- 35 Schuldt EZ, Farias MR, Ribeiro-do-Valle RM, Ckless K. Comparative study of radical scavenger activities of crude extract and fractions from Cuphea carthagenensis leaves. Phytomedicine 2004; 11: 523–9.
- 36 Huang D, Ou B, Prior RL. The chemistry behind antioxidant capacity assays. J Agric Food Chem 2005; 53: 1841–56.
- 37 Prior RL, Wu X, Schaich K. Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements. J Agric Food Chem 2005; 53: 4290–302.
- 38 Cho EJ, Yokozawa T, Rhyu DY, Kim SC, Shibahara N, Park JC. Study on the inhibitory effects of Korean medicinal plants and their main compounds on the 1,1-diphenyl-2-picrylhydrazyl radical. Phytomedicine 2003; 10: 544–51.
- 39 Yu J, Wang I, Walzem RL, Miller EG, Pike LM, Patil BS. Antioxidant activity of citrus limonoids, flavonoids, and coumarins. J Agric Food Chem 2005; 53: 2009–14.
- 40 Erdemoglu N, Turan NN, Cakici I, Sener B, Aydin A. Antioxidant activities of some Lamiaceae plant extracts. Phytother Res 2006; 20: 9–13.
- 41 Lee CP, Yen GC. Antioxidant activity and bioactive compounds of tea seed (Camellia oleifera Abel.) oil. J Agric Food Chem 2006; 54: 779–84.
- 42 Carbonari KA, Ferreira EA, Rebello JM, Felipe KB, Rossi MH, Felicio JD et al . Free-radical scavenging by Ouratea parviflora in experimentally-induced liver injuries. Redox Rep 2006; 11: 124–30.
- 43 Goupy P, Dufour C, Loonis M, Dangles O. Quantitative kinetic analysis of hydrogen transfer reactions from dietary polyphenols to the DPPH radical. J Agric Food Chem 2003; 51: 615–22.
- 44 Tsujimoto Y, Hashizume H, Yamazaki M. Superoxide radical scavenging activity of phenolic compounds. Int J Biochem 1993; 25: 491–94.
- 45 Hanasaki Y, Ogawa S, Fukui S. The correlation between active oxygens scavenging and antioxidative effects of flavonoids. Free Radic Biol Med 1994; 16: 845–50.
- 46 Pacher P, Beckman JS, Liaudet L. Nitric oxide and peroxynitrite in health and disease. Physiol Rev 2007; 87: 315–424.
- 47 Zou Y, Lu Y, Wei D. Antioxidant activity of a flavonoid-rich extract of Hypericum perforatum L. in vitro. J Agric Food Chem 2004; 52: 5032–9.
- 48 Joubert E, Winterton P, Britz TJ, Gelderblom WCA. Antioxidant and pro-oxidant activities of aqueous extracts and crude polyphenolic fractions of Rooibos (Aspalathus linearis). J Agric Food Chem 2005; 53: 10260–7.
- 49 Varga Z, Seres I, Nagy E, Ujhelyi L, Balla G, Balla J et al . Structure prerequisite for antioxidant activity of silybin in different biochemical systems in vitro. Phytomedicine 2006; 13: 85–93.
- 50 Rice-Evans C. Free radicals and antioxidants in normal and pathological processes. In: C Rice-Evans, KR Bruckdorfer (eds). Oxidative Stress, Lipoproteins and Cardiovascular Dysfunction. Portland Press, London, 1995; 1–32.
- 51 Stark G. Functional consequences of oxidative membrane damage. J Membr Biol 2005; 205: 1–16.
- 52 Kadiiska MB, Gladen BC, Baird DD, Germolec D, Graham LB, Parker CE et al . Biomarkers of oxidative stress study II. Are oxidation products of lipids, proteins and DNA markers of CCl4 poisoning? Free Radic Biol Med 2005; 38: 698–710.
- 53 Packer JE, Slater TF, Wilson RL. Reaction of the carbon tetrachloride related peroxy free radical with amino acids: pulse radiolysis evidence. Life Sci 1987; 23: 2620–7.
- 54 Escobar JA, Rubio MA, Lissi EA. SOD and catalase inactivation by singlet oxygen and peroxyl radicals. Free Radic Biol Med 1996; 20: 285–90.
- 55 Aniya Y, Anders MW. Alteration of hepatic glutathione-s-transferase and release into serum after treatment with bromobenzene and carbon tetrachloride. Biochem Pharmacol 1985; 39: 4239–44.
- 56 Farombi EO. Mechanisms for the hepatoprotective action of kolaviron: studies on hepatic enzymes, microsomal lipids and lipid peroxidation in carbon tetrachloride-treated rats. Pharmacol Res 2000; 42: 75–80.
- 57 Ha BJ, Lee JY. The effect of chondroitin sulfate against CCl4– induced hepatotoxicity. Biol Pharm Bull 2003; 26: 622–26.
- 58 Aniya Y, Koyama T, Miyagi C, Miyahira M, Inomata C, Kinoshita S et al . Free radical scavenging and hepatoprotective actions of the medicinal herb, Crassocephalum crepidioides from the Okinawa Islands. Biol Pharm Bull 2005; 28: 19–23.
- 59 Gao Z, Xu H, Chen X, Chen H. Antioxidant status and mineral contents in tissues of rutin and baicalin fed rats. Life Sci 2003; 73: 1599–607.
Citing Literature
