Chromatographic analysis, antiproliferative effect and genotoxicity of aqueous extracts of Citrus sinensis (L) osbeck on the Allium cepa L test system / Análise cromatográfica, efeito antiproliferativo e genotoxicidade de extratos aquosos de Citrus sinensis (L) osbeck sobre o sistema teste de Allium cepa L

The species Citrus sinensis (L.) Osbeck (orange) is used in folk medicine as an infusion (tea) for the treatment of nerve irritation, insomnia, spasms, whooping cough, flue, and gastrointestinal problems. The aim of this study was to evaluate the antiproliferative and genotoxic potential of aqueous extracts of C. sinensis testing by Allium cepa L., as well as to determine the components in the complex mixture (tea). Leaves from two populations of this species were used in the preparation of infusions at two concentrations: 6g.L-1 and 24g.L-1, besides using distilled water as a negative control and 9.6% glyphosate as a positive control. Six groups of four bulbs of A. cepa were used, each group corresponding to one of the treatments. The bulbs were rooted in distilled water and then transferred to the respective treatments where they remained for 24 hours (the bulbs of the negative control remained in distilled water). The rootlets were collected, fixated in ethanol-acetic acid (3:1) for 24 hours and stored in 70% alcohol. 4,000 cells were analyzed per group of bulbs, and the calculated mitotic indices were submitted to statistical analysis using the c 2 test. The aqueous extracts of C. sinensis have as main components flavonoids and phenolic acids. The values of the mitotic index decreased with increasing concentration of the extracts relative to the control in water. The results indicated that the orange aqueous extracts in the studied concentrations in both populations have antiproliferative and genotoxic effects on the cell cycle of A. cepa.

A espécie Citrus sinensis (laranjeira) (L.) Osbeck é usada na medicina popular como infusão (chá) para o tratamento de irritações nervosas, insônia, espasmos, tosse convulsa, gripes e problemas gastrointestinais. O objetivo desse estudo foi avaliar o potencial antiproliferativo e genotóxico dos extratos aquosos de Citrus sinensis pelo teste de Allium cepa L., bem como determinar os componentes na mistura complexa (chá). Folhas de duas populações dessa espécie foram utilizadas no preparo das infusões em duas concentrações: 6g.L-1 e 24g.L-1, além de serem usados água destilada como controle negativo e glifosato 9,6% como controle positivo. Utilizaram-se seis grupos de quatro bulbos de A. cepa, cada grupo correspondendo a um dos tratamentos. Os bulbos foram enraizados em água destilada e então transferidos para os respectivos tratamentos onde permaneceram por 24 horas (os bulbos do controle negativo permaneceram em água destilada). As radículas foram coletadas, fixadas em etanol-ácido acético (3:1) por 24 horas e armazenadas em álcool 70%. Foram analisadas 4,000 células por grupo de bulbos, e os índices mitóticos calculados submetidos à análise estatística pelo teste c 2. Os extratos aquosos de C. sinensis têm como componentes principais flavonoides e ácidos fenólicos. Os valores dos índices mitóticos diminuíram com o aumento das concentrações dos extratos, em relação ao controle em água. Os resultados indicaram que os extratos aquosos de laranjeira nas concentrações estudadas das duas populações possuem ação antiproliferativa e genotóxica sobre o ciclo celular de A. cepa.

Chromatographic analysis and antiproliferative potential of aqueous extracts of Punica granatum fruit peels using the Allium cepa test

Punica granatum L., locally known as romanzeira, is native to Asia but found throughout Brazil. P. granatum is used for treating inflammatory, infectious and respiratory diseases. The aim of this study was to evaluate the chromatography and genotoxicity of an aqueous extract of P. granatum (pomegranate) fruit peel using the Allium cepa L. test. The experiment set-up entailed 7 treatments: T1-distilled water, T2-tea 5 g.L-1, T3-tea 10 g.L-1, T4-glyphosate at 9.6%, T5-glyphosate with subsequent recovery in distilled water, T6-glyphosate with subsequent recovery in tea 5 g.L-1 and T7-glyphosate with subsequent recovery in tea 10 g.L-1. The rootlets were collected and fixed in ethanol:acetic acid (3:1) for 24 hours, then stored in 70% ethanol under refrigeration. Analysis was performed using high performance liquid chromatography for the quantification of the extracted phenolic compounds. Gallic acid, catechin, caffeic acid, and rutin were abundant in the extracts of P. granatum. The extracts were found to exhibit antiproliferative potential but not antimutagenic or genotoxic activity.

Punica granatum L., conhecida como romanzeira, é originária da Ásia e encontra-se distribuída por todo Brasil. É usada para o tratamento de doenças inflamatórias, infecciosas e respiratórias. Em decorrência da grande utilização de recursos fitoterápicos, é necessário esclarecer à população sobre a grande quantidade de substâncias existentes nas plantas e sobre os benefícios e prejuízos de tais substâncias à saúde. O presente trabalho objetivou realizar a análise cromatográfica e o estudo da genotoxicidade dos extratos aquosos das cascas dos frutos de P. granatum através do teste de Allium cepa L. Para a montagem do experimento, foram utilizados 7 tratamentos: T1-água destilada, T2-chá 5 g.L-1, T3-chá 10 g.L-1, T4-glifosato a 9,6%, T5-glifosato para recuperação em água destilada, T6-glifosato para recuperação em chá 5 g.L-1 e T7-glifosato para recuperação em chá 10 g.L-1. As radículas foram coletadas e fixadas em etanol:ácido acético (3:1) por 24 horas, e armazenadas em álcool 70%, sob refrigeração. Realizou-se análise por cromatografia líquida de alta eficiência para quantificação dos compostos fenólicos. Nos extratos de P. granatum foram observados em maior quantidade: ácido gálico, catequina, ácido cafeico e rutina. Além disso, os extratos demonstraram potencial antiproliferativo, sem apresentar atividade antimutagênica e genotóxica.

Genotoxic and chromatographic analyses of aqueous extracts of Peltodon longipes Kunth ex Benth. (hortelã-do-campo)

Peltodon longipes is used as a stimulant and emmenagogue. The objective of this study was to perform genotoxic and chromatographic analyses of the extracts of two samples of P. longipes, collected from the cities of Santa Maria and Tupanciretã, RS, Brazil. The Allium cepa assay was used to analyze genotoxicity while high-performance liquid chromatography was employed to determine phenolic compounds. The genotoxicity experiment consisted of nine groups each comprising four A. cepa bulbs. Bulb roots were developed in distilled water and then transferred for the treatments, for 24 hours, and the negative control remained in water. The treatments were: aqueous extracts at concentrations of 5 and 15 g L-1 for each sample, plus four groups treated with 1% glyphosate, one of which was used as a positive control and the other three for testing DNA damage recovery using water and the extracts of P. longipes from Santa Maria. All extracts of P. longipes exhibited anti-proliferative potential, although the effect was significantly greater for the extracts from the Tupanciretã sample. This sample also contained the highest amount of rosmarinic acid and kaempferol, which may confer the effects found in these extracts. Only extracts from the Santa Maria sample exhibited genotoxic potential.

Peltodon longipes é utilizada como estimulante e emenagoga. Objetivou-se realizar análises genotóxica e cromatográfica dos extratos de duas amostras de P. longipes, coletadas nos municípios de Santa Maria e Tupanciretã, RS, Brasil. O teste de Allium cepa foi utilizado para análise da genotoxicidade e a cromatografia líquida de alta eficiência, para determinação dos compostos fenólicos. O experimento de genotoxicidade constou de nove grupos de quatro bulbos de A. cepa. Os bulbos foram enraizados em água destilada e após transferidos para os tratamentos, por 24 horas, permanecendo o controle negativo em água. Os tratamentos foram: extratos aquosos nas concentrações de 5 e 15 g L-1 de cada amostra, além de quatro grupos tratados com glifosato 1%, um deles usado como controle positivo e outros três para testar a recuperação de danos ao DNA, utilizando água e os extratos de P. longipes da amostra de Santa Maria. Todos os extratos de P. longipes demonstraram potencial antiproliferativo, porém o efeito foi significativamente maior para os extratos da amostra de Tupanciretã. Essa amostra também apresentou maior quantidade de ácido rosmarínico e canferol, o que pode estar relacionado com os efeitos encontrados nesses extratos. Somente extratos da amostra de Santa Maria demonstraram potencial genotóxico.

Post-treatment with plant extracts used in Brazilian folk medicine caused a partial reversal of the antiproliferative effect of glyphosate in the Allium cepa test.

Species of the genus Psychotria are used for multiple purposes in Brazilian folk medicine, either as water infusions, baths or poultices. This study was aimed to evaluate the genotoxic and antiproliferative effects of infusions of Psychotria brachypoda and P. birotula on the Allium cepa test. Exposure to distilled water was used as a negative control, while exposure to glyphosate was used as a positive control. The interaction of extracts (as a post-treatment) with the effects of glyphosate was also studied. Results showed that glyphosate and the extracts of both P. brachypoda and P. birotula reduced the mitotic index as compared with the negative control (distilled water). Surprisingly, however, both extracts from P. brachypoda and P. birotula caused a partial reversal of the antiproliferative effect of glyphosate when used as a post-treatment. Glyphosate also induced the highest number of cells with chromosomal alterations, which was followed by that of P. birotula extracts. However, the extracts from P. brachypoda did not show any significant genotoxic effect. Post-treatment of glyphosate-treated samples with distilled water allowed a partial recovery of the genotoxic effect of glyphosate, and some of the Psychotria extracts also did so. Notably, post-treatment of glyphosate-treated samples with P. brachypoda extracts induced a statistically significant apoptotic effect. It is concluded that P. brachypoda extracts show antiproliferative effects and are not genotoxic, while extracts of P. birotula show a less potent antiproliferative effect and may induce chromosomal abnormalities. The finding of a partial reversion of the effects of glyphosate by a post-treatment with extracts from both plants should be followed up.

Post-treatment with plant extracts used in Brazilian folk medicine caused a partial reversal of the antiproliferative effect of glyphosate in the Allium cepa test. / Post-treatment with plant extracts used in Brazilian folk medicine caused a partial reversal of the antiproliferative effect of glyphosate in the Allium cepa test.

Species of the genus Psychotria are used for multiple purposes in Brazilian folk medicine, either as water infusions, baths or poultices. This study was aimed to evaluate the genotoxic and antiproliferative effects of infusions of Psychotria brachypoda and P. birotula on the Allium cepa test. Exposure to distilled water was used as a negative control, while exposure to glyphosate was used as a positive control. The interaction of extracts (as a post-treatment) with the effects of glyphosate was also studied. Results showed that glyphosate and the extracts of both P. brachypoda and P. birotula reduced the mitotic index as compared with the negative control (distilled water). Surprisingly, however, both extracts from P. brachypoda and P. birotula caused a partial reversal of the antiproliferative effect of glyphosate when used as a post-treatment. Glyphosate also induced the highest number of cells with chromosomal alterations, which was followed by that of P. birotula extracts. However, the extracts from P. brachypoda did not show any significant genotoxic effect. Post-treatment of glyphosate-treated samples with distilled water allowed a partial recovery of the genotoxic effect of glyphosate, and some of the Psychotria extracts also did so. Notably, post-treatment of glyphosate-treated samples with P. brachypoda extracts induced a statistically significant apoptotic effect. It is concluded that P. brachypoda extracts show antiproliferative effects and are not genotoxic, while extracts of P. birotula show a less potent antiproliferative effect and may induce chromosomal abnormalities. The finding of a partial reversion of the effects of glyphosate by a post-treatment with extracts from both plants should be followed up.