Print version ISSN 0102-695X
Rev. bras. farmacogn. vol.17 no.1 João Pessoa Jan./Mar. 2007
Bioatividade em Artemia salina de várias espécies de Solanum do Nordeste Brasileiro
Tania Maria S. SilvaI, *; Roberto Jefferson B. NascimentoII; Michelle M. BatistaII; Maria F. AgraII; Celso A. CamaraIII
INúcleo Complexo Produtivo de Saúde, Instituto Multidisciplinar em Saúde, Campus Avançado Anísio Teixeira, Avenida Olívia Flores, 3000, Candeias, 45055-090, Vitória da Conquista, BA, Brazil,
IILaboratório de Tecnologia Farmacêutica, Universidade Federal da Paraíba, Caixa Postal 5009, 58051-970, João Pessoa, PB, Brazil,
IIIDepartamento de Química, Universidade Federal Rural de Pernambuco, R. Dom Manoel de Medeiros, s/n, Dois Irmãos, 52171-900, Recife, PE, Brazil
Os extratos metanólicos de 13 espécies de Solanum (Solanaceae) foram testados para verifificação da bioatividade em Artemia salina. As espécies testadas (partes aéreas, raízes e frutos) foram: S. asperum, S. capsicoides, S. palinacantum, S. paludosum, S. paniculatum, S. paraibanum, S. sisymbriifolium, S. crinitum, S. diamantinense, S. megalonyx, S. torvum, S. asterophorum e S. stipulaceum. Das treze espécies testadas, quatro foram inativas. Os extratos dos frutos de S. asperum (CL50 = 420,5 µg/mL) e S. paludosum (CL50 = 548,0 µg/mL), partes aéreas de S. diamantinense (CL50 = 481,0 µg/mL) e S. sisymbrifolium (CL50 = 382,7 µg/mL), e das raízes S. asperum (CL50 = 593,4 µg/mL) e S. stipulaceum (CL50 = 823,1 µg/mL) que mostraram atividade moluscicida contra Biomphalaria glabrata também mostraram atividade tóxica em Artemia salina.
Unitermos: Solanaceae, Solanum, Artemia salina, bioatividade.
The methanolic extracts of 13 Specieis of the genus Solanum (Solanaceae) have been tested for bioactivity in Artemia salina. The extracts investigated were prepared from various parts (aerial parts, roots and fruits) of S. asperum, S. capsicoides, S. palinacantum, S. paludosum, S. paniculatum, S. paraibanum, S. sisymbriifolium, S. crinitum, S. diamantinense, S. megalonyx, S. torvum, S. asterophorum and S. stipulaceum. The lethal concentrations were determined for the extracts and among thirteen plants tested, four appear to be inactive. The extracts of the fruits of S. asperum (LC50 = 420.5 µg/mL) and S. paludosum (LC50 = 548.0 µg/mL), aerial parts of S. diamantinense (LC50 = 481.0 µg/mL) and S. sisymbrifolium (LC50 = 382.7 µg/mL), and the roots of S. asperum (LC50 = 593.4 µg/mL) and S. stipulaceum (LC50 = 823.1 µg/mL), all of which previously showed molluscicidal activity against Biomphalaria glabrata were also found to be active in the present study with brine shrimp.
Keywords: Solanaceae, Solanum, Artemia salina, bioactivity, brine shrimp.
The genus Solanum is considered to be one of the largest and most complex among the Angiosperms. It is comprised of about 1500 species and has at least 5000 published epithets (Nee, 1999). The genus is well represented in Brazil and is widely distributed from north to south in diverse phytogeographic regions. Many of the species are endemic to the country and well represented in the northeast Brazil with about 80 species which are widely distributed in the region. About 20 of these Solanum species are endemic to the northeastern region (Agra, 1999) and are widely used in folk medicine, commonly known as "jurubeba"; the name derived from the Tupi-guarani word "yubeba", which refers to the presence of prickles in some of them.
The presence of the steroidal alkaloid solasodine, which is potentially an important starting material for the synthesis of steroid hormones, is characteristic of the genus Solanum (Silva et al., 2005a; Silva et al., 2005b; Barbosa-Filho et al., 1991).
The brine shrimp assay has been established as a safe, practical, and economic method for the determination of the bioactivity of synthetic compounds (Almeida et al., 2002) as well as plant products (Meyer et al., 1982; McLaughlin et al., 1991; Lhullier et al., 2006; Stefanello et al., 2006). The significant correlation between the brine shrimp assay and in vitro growth inhibition of human solid tumor cell lines demonstrated by the National Cancer Institute (NCI, USA) is significant because it shows the value of this bioassay as a pre-screening tool for antitumor drug research (Anderson et al., 1991). The wide distribution, anticancer properties (Kupchan et al., 1965; Cham et al., 1987; Cham, 1994; Daunter; Cham, 1990; Hu et al., 1999; Esteves-Souza et al., 2002; Lee et al., 2004; Friedman et al., 2005) of the glycoalkaloids and molluscicidal activity (Silva et al., 2005c) of the crude extracts, led us to the selection of these Solanum species for the present work.
MATERIAL AND METHODS
All together, 13 species of the genus Solanum were investigated in our present study. All species were collected in Northeastern Brazil and identified by M. F. Agra, the Head of the Botany Section of LTF-UFPB. Voucher specimens (Table 1) were deposited at the Prof. Lauro Pires Xavier Herbarium (JPB) and the duplicates were kept in the collection of references at LTF, both at the Universidade Federal da Paraíba, João Pessoa, Brazil.
Preparation of the crude extract
The extracts for the present study for bioassay against Artemia salina were prepared as follows: The dried and powdered plant materials such as, fruits of S. asperum, S. capsicoides, S. palinacantum, S. paludosum, S. paniculatum, S. paraibanum and S. sisymbriifolium, aerial parts of S. asperum, S. asterophorum, S. capsicoides, S. crinitum, S. diamantinense, S. megalonyx, S. palinacantum, S. paniculatum, S. sisymbriifolium, and S. torvum, and roots of S. asperum, S. asterophorum, S. palinacantum, S. paludosum and S. stipulaceum were extracted with methanol at room temperature followed by treatment as described before (Silva et al., 2005c) for the preparation of the test samples for molluscicidal activity.
The brine shrimp lethality assay was performed following the reported procedure (Meyer, 1982) with some modifications (Silva et al, 2005d). The growth medium was prepared with sea water in a small tank divided into two compartments. The shrimp eggs were added to the covered compartment. A lamp was placed above the open side of the tank to attract hatched shrimps through perforations in the partition wall. After 48 h, the shrimps mature as nauplii (A. salina) and are ready for the assay. Test extracts were dissolved in three drops of Cremophor®, 2 mL of DMSO and sea water to complete 5 mL of total volume. Appropriate volumes of the resulting solution were then added in tubes, in quadruplicate, with 5 mL of saline solution containing 10 nauplii each to afford the final sample concentrations. The control samples containing Cremophor® and DMSO, under the same conditions, do not cause significant brine shrimp mortality. After 24 h incubation under light, the number of dead and survivor brine shrimps in each tube was counted. The LC50 values were calculated by graphics from drug concentration vs. lethality percentage using a probit scale adjust. Data analysis was performed with Origin 6.0 software.
RESULTS AND DISCUSSION
Many of the collected species for the present study are already used in folk medicine. The roots of S. asterophorum, S. paniculatum and S. torvum, for example, are used in the treatment of liver diseases (Agra; Bhattacharrya, 1999), and this appears to be its most common ethnomedicinal use. A wine is produced commercially from the fruits of S. paniculatum, which also have use in popular medicine. Solanum asperum can cause skin irritation (Agra; Battacharyya, 1999). The ethnomedicinal data and other uses of the studied Solanum species are given in Table 1. The plants known to be toxic in popular medicine also showed brine shrimp bioactivity in our study.
The brine shrimp lethality for different extracts of Solanum species are given in Table 2. The extracts are considered inactive when all nauplii survive at a concentration of 1000 µg/mL (Meyer 1982). Among the thirteen plants tested, four seems to be inactive. As stated in Table 2, the extracts demonstrating molluscicidal activity (Silva et al., 2005c) were also found to be active in the brine shrimp assay. The aerial parts of S. asperum (entry 1) and S. megalonyx (entry 10), and the roots of S. palinacanthum (entry 13) were found to be inactive in both molluscicidal and brine shrimp bioassays.
The data in Table 2 show that many Solanum species (S. asterophorum, S. capsicoides, S. crinitum, S. palinacanthum, S. paniculatum, S. sisymbrifolium, and S. torvum) which are active in brine shrimp bioassay are not active against the mollusk Biomphalaria glabrata. On the other hand, all the Solanum species which demonstrated molluscicidal activity (Silva et al., 2005c) were also active against A. salina. The most active extracts in this study were found to be the ones from the roots of S. asterophorum (CL50 = 107.3 µg/mL; entry 5) and from the aerial parts of S. torvum with CL50 = 295.2 µg/mL (entry 22). Surprisingly, the chemical study of the latter species did not show the presence of any glycoalkaloid (Mahmood et al., 1985), whereas, the former showed the presence of alkaloids (Silva et al., 2005c). This suggests that the observed bioactivity is not only due to the presence of alkaloids but other constituents (e.g., saponins, sapogenins) may also play an important role.
The molluscicidal activity seen in some Solanum species is generally attributed to the presence of glycoalkaloids. The other classes of secondary metabolites, including alkamines, have little if any activity (Alzerreca; Hart, 1982; Wanyonyi et al., 2003; Silva et al., 2005c). Our study, however, showing bioactivity in the brine shrimp assay for some extracts with no corresponding molluscicidal activity suggests that other classes of secondary metabolites must be involved in the process, and, therefore, other screening (e.g., anticancer, antifungical) must be performed on such extracts.
The authors thank CNPq, CAPES, IMSEAR-CNPq, and PIBIC-UFPb for scholarships and financial support, and Prof. J. Bhattacharyya from Laboratório de Tecnologia Farmacêutica-UFPB for useful help in the preparation of the manuscript.
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Received 05/05/06. Accepted 11/28/06