Antioxidant, Antibacterial, Cytotoxic, and Anti-Inflammatory Potential of the Leaves of Solanum Lycocarpum A

Hindawi Publishing Corporation Evidence-Based Complementary and Alternative Medicine Volume 2015, Article ID 315987, 8 pages http://dx.doi.org/10.1155/2015/315987 Research Article Antioxidant, Antibacterial, Cytotoxic, and Anti-Inflammatory Potential of the Leaves of Solanum lycocarpum A. St. Hil. (Solanaceae) Guilherme Augusto Ferreira da Costa, Melissa Grazielle Morais, Aline Aparecida Saldanha, Izabela Caputo Assis Silva, Álan Alex Aleixo, Jaqueline Maria Siqueira Ferreira, Adriana Cristina Soares, Joaquim Maurício Duarte-Almeida, and Luciana Alves Rodrigues dos Santos Lima Universidade Federal de Sao˜ Joao˜ Del-Rei, Campus Centro-Oeste Dona Lindu, 35501-296 Divinopolis,´ MG, Brazil Correspondence should be addressed to Luciana Alves Rodrigues dos Santos Lima; [email protected] Received 18 March 2015; Accepted 20 April 2015 Academic Editor: Daniela Rigano Copyright © 2015 Guilherme Augusto Ferreira da Costa et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. EthanolextractandfractionsobtainedfromleavesofSolanum lycocarpum were examined in order to determine their phenolic composition, antioxidant, antibacterial, anti-inflammatory, and cytotoxic potential. High performance liquid chromatography coupled with DAD analysis indicated that the flavonoids apigenin and kaempferol were the main phenolic compounds present in dichloromethane and ethyl acetate fractions, respectively. The antioxidant activity was significantly more pronounced for dichloromethane, ethyl acetate, and hydroethanol fractions than that of the commercial antioxidant 2,6-di-tert-butyl-4- methylphenol. The hexane and dichloromethane fractions were more active against the tested bacteria. The hydroethanol fraction exhibited significant anti-inflammatory activity at the dose of 75 and 150 mg/kg in the later phase of inflammation. However, the antiedematogenic effect of the higher dose of the ethyl acetate fraction (150 mg/kg) was more pronounced. The ethyl acetate fraction also presented a less cytotoxic effect than the ethanol extract and other fractions. These activities found in S. lycocarpum leaves can be attributed, at least in part, to the presence of phenolic constituents such as flavonoids. This work provided the knowledge of phenolic composition in the extract and fractions and the antioxidant, antibacterial, anti-inflammatory, and cytotoxic activities of leaves of S. lycocarpum. 1. Introduction rhamnosides [5, 6]. Cinnamic acids, such as caffeic and chlorogenic acids, are also very well known for their antioxi- Polyphenolic compounds, such as flavonoids and phenolic dant properties [7, 8]. acids, are commonly consumed as natural components of The Solanaceae family comprises about 3000 species and vegetables, beans, fruits, and/or phytotherapeutics and are 150 genera. It is prevalent in tropical and subtropical regions generally regarded as safe chemicals, thus displaying low toxi- of South America and has economic importance because cological activity [1]. These compounds have been presented several species of the Solanum genus are cultivated for food, as therapeutic alternatives for the treatment of a large number such as Solanum tuberosum (potato), Solanum lycopersicum of pathologies [2]. Flavonoids, particularly apigenin and (tomato), Solanum melongena (eggplant), and Solanum gilo luteolin, have shown potent activity against oxidative stress (gilo) [9]. The species Solanum lycocarpum A. St. Hil., pop- [3]. Other studies in several systems have revealed that ularly known as the “fruit of the wolf,” is widely distributed apigenin was able to inhibit tumor growth [4]. Additionally, in the Brazilian Cerrado. The fruits are usually consumed “in recent studies have shown anti-inflammatory, antimicrobial, natura” or used in jellies, jams, or pasta preparations [10]. It and antioxidant properties for kaempferol and kaempferol is widely used in traditional medicine as a sedative, in the 2 Evidence-Based Complementary and Alternative Medicine treatment of epilepsy, asthma, diabetes, obesity, the reduction modifications. Exactly 2 mL of 2% aluminum trichloride of cholesterol levels, and abdominal and renal pains [11]. (AlCl3) in methanol was mixed with the same volume of the Biological activities such as anti-inflammatory, antioxidant, extract or fraction solution (1.0 mg/mL). The absorbance was cytotoxic, antibacterial, allelopathic, and larvicidal have been read at 415 nm using a Hitachi 2010 spectrophotometer after reported for this species [10, 12–14]. 10 min, with a blank sample consisting of a 2 mL extract or Phytochemical studies of this species showed that sola- fraction solution with 2 mL methanol without AlCl3.Rutin sonine and solamargine are two predominant steroidal gly- was used as the reference compound to produce the standard coalkaloids, which have a common aglycone, solasodine [15]. curve, and total flavonoid contents were expressed as gof A recent study also reported the presence of cinnamic acids, rutin equivalents/mg of extract or fraction. All assays were such as caffeic and chlorogenic acids, in ripe fruits of S. performed in triplicate. lycocarpum [13]. However, until now, no data exist about the phenolic composition of the leaves of S. lycocarpum 2.4. Phenolic Profiles by HPLC-DAD. The phenolic sub- and their antioxidant, antibacterial, anti-inflammatory, and stances in the ethanol extract and fractions were carried cytotoxic activities. The aim of this study was to identify the out using analytical reversed phase HPLC on an Agilent main phenolic compounds and estimate the total flavonoid 1260 system with an autosampler and quaternary pump content of the ethanol extract (EE) and fractions obtained coupled with a diode array detector. Compound separation from the leaves of S. lycocarpum and also to demonstrate their was performed by a Zorbax Eclipse Plus 5B RP-18 (5 m, 250 antioxidant, antibacterial, anti-inflammatory, and cytotoxic × 4.6 mm, Agilent, USA) column. The mobile phases were potential. (A) water/formic acid (99.9 : 0.1) and (B) acetonitrile. The gradient consisted of 20% of (B) for 2 minutes and increased 2. Materials and Methods to 30% of (B) after 10 minutes, 50% of (B) after another 10 minutes, and 70% of (B) after an additional 10 minutes. 2.1. Chemicals. Ascorbic acid (AA), 2,6-di-tert-butyl-4- Forcolumncleaning,90%ofphase(B)wasused[7]. Each methylphenol (BHT), 1,1-diphenyl-2-picrylhydrazyl (DPPH), samplewasinjectedinduplicate,rangingfrom5to20L kaempferol, quercetin, rutin, apigenin, luteolin, coumaric according to the sample concentration. Determination was acid, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium performed by comparing the retention times and the UV bromide (MTT), carrageenan type IV, indomethacin, spectra against those obtained from the standards (apigenin, HPLC grade methanol, and acetonitrile were obtained from kaempferol, luteolin, quercetin, rutin, caffeic, coumaric, fer- Sigma (St. Louis, MO, USA). Caffeic, ferulic, and chlorogenic ulic, and chlorogenic acids). The determination was positive acids were purchased from Apin Chemicals Ltd. (Abingdon, when the similarity between the chromatograms was equal to UK). Mueller Hinton broth (MH) medium was acquired or greater than 90%. from Himedia (Mumbai, India). Streptomycin, penicillin, and fetal bovine serum (FBS) were purchased from Cultilab 2.5. DPPH Radical Scavenging Assay. The radical scavenging (Campinas, SP, Brazil). Dulbecco’s Modified Eagle’s Medium abilities of extract and fractions of S. lycocarpum were (DMEM) was purchased from Gibco (Grand Island, NY, based on reactions with 1,1-diphenyl-2-picrylhydrazyl radical USA). All other reagents were of analytical grade and were (DPPH) and compared to standards, 2,6-di-tert-butyl-4- obtained from Vetec (Duque de Caxias, RJ, Brazil). methylphenol (BHT) and ascorbic acid (AA). Determination of antioxidant activity by the DPPH method was adapted 2.2. Plant Material and Extraction. The leaves of S. lyco- for use with microplates [17]. Briefly, a solution of DPPH carpum A.St.Hil.werecollectedinSao˜ Sebastiao˜ do Oeste, (0.002% w/v) was prepared in 80% methanol. Volumes of Minas Gerais, Brazil, in August 2011. The plant material 75 L of samples or standards (1, 10, 100, 250, and 500 g/mL) was identified by Dr. Alexandre Salino and a voucher were added to wells in a 96-well flat-bottom plate containing specimen (BHCB 159397) was deposited at the Instituto 150 LofDPPHsolution.Theplatewasthencoveredand ∘ de Cienciasˆ Biologicas´ Herbarium, Universidade Federal de left in the dark at room temperature (25 C). After 30 min, Minas Gerais, Belo Horizonte, MG, Brazil. As S. lycocarpum absorbance at 517 nm was measured in a spectrophotometer a Brazilian native genetic material, the present study had been (Biotek Power Wave XS2/US), and 80% methanol was used approved by the Conselho Nacional de Desenvolvimento for the baseline correction. Scavenging ability was expressed Cient´ıfico e Tecnologico´ (CNPq) through Access and Ship- astheinhibitionpercentageandwascalculatedbythe ment Component of Genetic Heritage for scientific research following equation [18]: purpose (number 010655/2011-5). Extraction of the dried and powdered leaves (121.00 g) by Scavenging ability (%) percolation (EtOH, 7 L, 72 h) gave the EE (17.02 g). Part of this extract (7.90 g) was dissolved in EtOH/H2O (1 : 1) and succes- (Abs − Abs ) (1) = control sample × 100,

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