ABSTRACT
Cissus verticillata and Sphagneticola trilobata have been used in Brazilian folk medicine for Diabetes Mellitus treatment, although their pharmacological and toxicological profile has not been clearly established. Thus, the aim of this study was to evaluate the preclinical toxicity of the aqueous extracts of C. verticillata and S. trilobata. The main groups of secondary metabolites were investigated, and the species differed by the presence of coumarins in C. verticillata and by tannins in S. trilobata extracts. The highest contents of phenolic compounds and flavonoids were quantified in C. verticillata infusion with 2.594 ± 0.04 mg equivalents of gallic acid g-1 of extract and 1.301 ± 0.015 mg equivalents of catechin g-1 of extract, respectively. While the extract of S. trilobata showed minimum values of these compounds, with 0.002 ± 0.001 mg equivalents of gallic acid g-1 extract and 0.005 ± 0.0004 mg equivalents of catechin g-1 of extract, respectively. These differences implied the results of in vitro antioxidant activity evaluated using ferric reducing antioxidant power (FRAP), in which the sample of C. verticillata at 5 mg mL-1 showed a value of 122 µM ferrous sulfate equivalents (FSE), while S. trilobata showed 0.93 µM FSE at the same concentration. With respect to cytotoxic assay with murine fibroblast cell line (3T3) only S. trilobata exhibited cytotoxic effects measured by MTT and Sulforhodamine B assays, evidenced by the cell viability value of approximately 16%, in both tests after 24 and 72 hours of exposure of the cells to 5 mg mL-1 of the extract. Comparatively, at 5 mg mL-1 the C. verticillata extract showed cell viability of 142% and 95%, respectively, after 24 hours of cell exposure. On the other hand, both species showed genotoxic profiles evidenced by chromosomal aberrations by Allium cepa bioassay, observed by the higher percentage values of chromosome bridges, chromosome loss, and disturbed anaphase for all concentrations of both extracts than those of the negative control. The results support the characterization of the toxicological profile for both species and create an alert regarding the use of S. trilobata, which should be avoided.
Subject(s)
Asteraceae/cytology , Asteraceae/chemistry , Asteraceae/toxicity , Diabetes Mellitus/diagnosis , Diabetes Mellitus/drug therapy , Vitaceae/cytology , Vitaceae/chemistry , Vitaceae/toxicityABSTRACT
A recent reinvestigation of aerial parts of Wedelia paludosa D.C. is described and reports, for the first time, the isolation of iso-kaurenoic acid from this species.
Uma recente reinvestigação das partes aéreas de Wedelia paludosa D.C. é descrita e relata, pela primeira vez, o isolamento do ácido iso-caurenóico desta espécie.
Subject(s)
Diterpenes/isolation & purification , Plant Extracts/isolation & purification , Wedelia/chemistry , Diterpenes/chemistry , Diterpenes/pharmacology , Plant Extracts/chemistry , Plant Extracts/pharmacologyABSTRACT
O presente trabalho teve por objetivo o estabelecimento de parâmetros para análise morfo-anatômica das partes aéreas (caule e folhas) da planta Wedelia paludosa DC., Asteraceae. A droga vegetal e o respectivo pó foram submetidos às análises macro e microscópica após preparação de lâminas semi-permanentes de cortes transversais e paradérmicos. O pó foi analisado após diafanização com cloral hidratado. É uma planta herbácea com caule castanho-avermelhado, folhas opostas, curto-pecioladas e membranáceas, apresentando pêlos nas duas faces, estreitada em direção à base acima do meio provida de dois pequenos lobos laterais e um terminal, maior e denteado. A planta apresenta secção circular de caule, que em crescimento primário, possui epiderme, colênquima angular, parênquima cortical e cilindro vascular, assim como oxalato de cálcio (várias drusas e alguns cristais prismáticos). A folha é simples, contendo tricomas tectores e estômato anomocítico. O mesofilo possui organização dorso-ventral e a nervura central, em corte transversal, apresenta-se côncavo-convexo com três feixes colaterais e presença de drusas de oxalato de cálcio nas células parenquimáticas. No pó obtido da droga vegetal foram verificadas as mesmas estruturas microscópicas observadas na droga vegetal rasurada. Os parâmetros morfo-anatômicos estabelecidos neste trabalho permitem o controle de qualidade macro e microscópico da planta tanto rasurada como na forma de pó.
This study aimed to establish parameters for morphoanatomical analysis of the aerial parts (stem and leaves) of the plant Wedelia paludosa DC., Asteraceae. The drug and its respective powder were characterized by macro and microscopic analysis after obtaining semi-permanent lamina of transverse and paradermal sections. The powder was analyzed after being cleared with chloral hydrate. It is an herbaceous plant with reddish-brown stems, opposite leaves, short-petiolated and membranous, with hair on both faces, narrowed towards the base up the middle provided with two small lateral lobes and a, large and jagged terminal. The plant has circular cross-section of stem, in which the primary growth, has epidermis, angular collenchyma, cortical parenchyma and vascular cylinder, as well as calcium oxalate (several clusters and some prismatic crystals). The leaf is simple, containing trichomes and stomata. The mesophyll has dorsiventral organization and the midrib, in cross section presents concave-convex face with three bundles and presence of calcium oxalate in parenchymal cells. In the powder was found the same structures observed in microscopic drug erased. The morphoanatomical parameters established in this work allow the macro and microscopic quality control of power and sectioned drug.
ABSTRACT
Wedelia paludosa D.C. (Asteraceae) is an ornamental species occurring in many regions of Brazil. Aiming to find new cytotoxic compounds, the hydromethanol extract of W. paludosa (HME), as well as the dichloromethane (DF) and water (WF) fractions resulting from its partition, were submitted to the brine shrimp lethality bioassay (BSLB) in order to evaluate their cytotoxicity. Dichloromethane fraction (DF) was shown to be the most cytotoxic fraction (LC50 = 140.6 μg/mL), and its analysis by reversed phase high performance liquid chromatography (RP-HPLC) revealed ent-kaurenoic (1, 6.22 ± 0.23 percent) and grandiflorenic (2, 3.22 ± 0.31 percent) acids as important constituents. HME (LC50 = 980 μg/mL), DF (LC50 = 140.6 μg/mL), 1 (LC50 = 15.9 μg/mL) and 2 (LC50 = 29.8 μg/mL) were found to be cytotoxic, while the water fraction (WF, LC50 >> 1000 μg/mL) was inactive. As conclusion, the cytotoxicity observed for HME and DF is mainly due to the presence of 1 and 2 in their constitution.
Wedelia paludosa D.C. (Asteraceae) é uma planta ornamental facilmente encontrada em várias regiões do Brasil, principalmente nos estados de Santa Catarina, São Paulo, Minas Gerais, Bahia e Pernambuco. Objetivando descobrir novas substâncias citotóxicas a partir desta espécie, o extrato hidrometanólico de W. paludosa (HME) e as frações diclorometânica (FD) e aquosa (FA) resultantes de sua partição em CH2Cl2-H2O foram avaliados utilizando-se o bioensaio em Artemia salina. A fração diclorometânica (FD) apresentou a maior atividade citotóxica (CL50 = 140,6 μg/mL), e sua análise por cromatografia líquida de alta eficiência empregando-se fase reversa (FR-CLAE) revelou os ácidos caurenóico (1, 6,22 ± 0,23 por cento) e grandiflorênico (2, 3,22 ± 0,31 por cento) como constituintes majoritários. As amostras HME (CL50 = 980 μg/mL), FD (CLC50 = 140,6 μg/mL), 1 (CL50 = 15,9 μg/mL) e 2 (CL50 = 29,8 μg/mL) foram citotóxicas contra A. salina, enquanto que a fração aquosa (FA, CL50 >> 1000 μg/mL) mostrou-se inativa. Conclui-se que a citotoxidade observada para HME e FD pode ser atribuída à presença dos ácidos caurenóico (1) e grandiflorênico (2) nestes extratos.
ABSTRACT
There are several reports of biological activity for kaurenoic acid (1), which is found in the aerial parts of Wedelia paludosa D.C., a herbaceous plant of the Asteraceae family abundant in Brazil, and that contains grandiflorenic acid (2) also. Both of these diterpenes were shown to cause lysis of trypomastigotes of the protozoa Trypanosoma cruzi, the causative agent of Chagas' disease (American trypanosomiasis). This paper reports the quantitative determination of these diterpene acids in the aerial parts of W. paludosa by an isocratic RP-HPLC method employing 60 percent acetonitrile in water and UV detection (220 nm). Kaurenoic acid (1) was found to be more abundant (0.85 ± 0.08 percent) while the amount of grandiflorenic acid (2) determined was almost three times lower (0.32 ± 0.02 percent). No significant seasonal variation was observed for these compounds in W. paludosa growing in Belo Horizonte (MG, Brazil) what points out to the aerial parts of this species are a good source of these biologically active diterpenes along all the year.