RESUMO
Piper amalago L. is used in Brazilian traditional medicine to treat inflammation, chest pain, and anxiety. This study aimed to investigate the safety and the renal and cardiovascular effects of the volatile oil (VO) and the aqueous (AE) and hydroalcoholic (HE) extracts from P. amalago. The gas chromatography-mass spectrometry analyses identified 47 compounds in the VO, with ß-cyclogermacrene, spathulenol, ß-phellandrene, and α-pinene standing out. Among the 47 compounds also found in AE and HE by liquid chromatography-mass spectrometry, glycosylated flavones, organic acids, amino acids, and amides were highlighted. Some examples of these compounds are methoxy-methylenedioxy cis-cinnamoyl pyrrolidine, methoxy-methylenedioxy trans-cinnamoyl pyrrolidine, and cyclobutene-2,4-bis-(1,3-benzodioxol-5-methoxy-6-yl)-1,3-dicarboxapyrrolidide. The acute toxicity experiments were conducted on female rats (n = 5). The cardiorenal assays (n = 8) and evaluations of vasodilatory effects on the mesenteric vascular bed (n = 5) were conducted on male rats. In either extract or VO, there were no mortality or changes in relative weights or histopathological analysis of the organs. Urinary volume and renal electrolyte excretion were elevated significantly during repeated dose 7-day treatment with different preparations from P. amalago. None of the preparations induced hypotension or changes in cardiac electrical activity. Only HE promoted significant vasodilatory effects in rats' isolated mesenteric vascular beds. These effects were completely abolished in the presence of L-NAME plus 4-aminopyridine. Therefore, P. amalago leaves are safe and present diuretic activity after acute and repeated dose administration over 7 days. Moreover, the HE induced significant vasodilator response in rats' mesenteric vascular beds by NO/cGMP pathway and voltage-dependent K+ channels activation.
RESUMO
Piper regnellii is a plant popularly known as "Pariparoba" and it is widely used in folk medicine to treat pain, inflammation, among others. This work presents the extraction, purification and characterization of polysaccharides present in the plant leaves and evaluation of their anti-inflammatory and antinociceptive activities. From the crude aqueous extract of P. regnellii leaves, a polysaccharide fraction named PR30R, predominantly constituted of arabinose, galactose and galacturonic acid monosaccharide units, was obtained. Methylation and NMR analysis showed that the main polysaccharides of PR30R are a type II arabinogalactan, formed by a ß-D-Galp-(1 â 3) main chain, substituted at O-6 by side chains of ß-D-Galp-(1 â 6), which are substituted at O-3 by non-reducing α-L-Araf ends, and a homogalacturonan, formed by â4)-α-D-GalpA-(1â units. Intraperitoneal administration of the crude polysaccharide fraction PRSF reduced significantly nociception induced by acetic acid in mice at the doses tested, and the PR30R fraction, derived from PRSF, presented antinociceptive and anti-inflammatory effects at a dose of 0.1096 mg/kg (PRSF ED50). These data support the use of the plant leaves in folk medicine as an herbal tea to treat pain and inflammation.
Assuntos
Piper , Animais , Camundongos , Polissacarídeos/farmacologia , Polissacarídeos/uso terapêutico , Polissacarídeos/análise , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Anti-Inflamatórios/análise , Inflamação , Analgésicos/farmacologia , Analgésicos/uso terapêutico , Folhas de Planta/química , Dor/tratamento farmacológicoRESUMO
Piper regnellii (Miq.) C. DC. var. regnellii, popularly known as "pariparoba", is used for therapeutic purposes. However, the morphological similarity among Piper species can lead to misidentifications, so molecular markers have been used to validate the identification of species and their morphotypes. Among the molecular markers used in plants, the intergenic spacer region trnL-trnF has proven effective in identifying plant species. For this reason, this region was used to evaluate two morphotypes of Piper regnellii var. regnellii. Studies with the trnL-trnF region have shown this region as a good marker for establishing phylogenetic relationships, distinguishing species, and identifying new species. We concluded that the trnL-trnF sequenced region show one indel of difference between the two morphotypes. It would be interesting to analyze these two morphotypes with a more variable region than the one used here, aiming to show intraspecific differences.(AU)
Assuntos
Filogenia , PiperRESUMO
ABSTRACT This study describes a comparative analysis of the essential oil (EO) chemical composition of leaves and reproductive organs (inflorescences and fruits) of Piper gaudichaudianum during the seasons of a year in order to determine the best collection time and the most suitable plant organ to obtain this extractive. The chemical composition of EO obtained from fresh leaves was compared to the dried ones, to verify if the drying process interferes in the extractive quality. The leaves were collected from a native population of Santa Maria, RS, Brazil, twice in each season, in triplicate, while inflorescences and fruits were sampled when they were present. The EO was obtained by hydrodistillation of the different plant organs for 3 h. The 20 EO samples were analyzed by gas chromatography (GC) coupled to mass spectrometry and GC with flame ionization detector, in triplicate. Hierarchical cluster analysis (HCA) and principal components analysis (PCA) were performed to verify a possible formation of chemical groups (CG) and the cohesion among them. The phenylpropanoid dillapiole was the major constituent of the EO in all seasons and in all plant organs, and myristicin was observed only in reproductive organs. The EO samples of this population were divided into two CG by HCA and PCA, showing the variability in chemical composition between different plant organs, however there was no chemical variability due to seasonality and phenophases. Since the drying of the leaves did not alter the EO chemical composition, this post-harvest procedure can be used without compromising the extrative quality.
RESUMO
O objetivo desse trabalho foi analisar a estrutura genética de populações de Pothomorphe umbellata (L.) Miq. com base em polimorfismos moleculares do tipo RAPD. Foram analisadas quatro populações naturais do estado de São Paulo (Jacareí, Jundiaí, Piquete e Ubatuba) e uma população do Paraná (Adrianópolis). Foram identificados 25 locos polimórficos (96,15%). Elevados índices de diversidade genética foram observados dentro das populações (Hs = 0,2220). Verificou-se que 65,33% da variabilidade genética total encontra-se dentro das populações e 34,67% entre as populações; índices estes, obtidos a partir do cálculo da divergência genética (G ST = 0,3467). Os resultados sugerem que essas populações possuem níveis elevados de variabilidade genética, a qual pode ser fortemente impactada pela ação humana.
The aim of this study was to analyze the genetic structure of populations of Pothomorphe umbellata (L.) Miq. based on RAPD molecular polymorphisms. Analysis included four natural populations from São Paulo State (Jacareí, Jundiaí, Piquete, Ubatuba) and one population from Paraná State (Adrianópolis). Twenty-five polymorphic loci (96.15%) were identified. There were high levels of genetic diversity within populations (Hs = 0.2220). Of the total genetic variability, 65.33% is within populations and 34.67% among populations (G ST = 0.3467). Results suggest that these populations have high levels of genetic variability, which can be strongly impacted by human action.
Assuntos
Variação Genética/genética , Piperaceae/crescimento & desenvolvimento , Estruturas Genéticas/genética , Plantas Medicinais/classificaçãoRESUMO
A exposição à radiação ultravioleta promove o desenvolvimento de efeitos indesejáveis, incluindo o fotoenvelhecimento e fotocarcinogênese. Uma forma de prevenção é a utilização de antioxidantes de origem natural. O extrato de raiz de Pothomorphe umbellata (PU) apresenta conhecida atividade antioxidante e fotoprotetora, atividade esta atribuída, em parte, ao princípio ativo 4-nerolidilcatecol (4-NC). Artigos publicados sobre a atividade do extrato de raiz de PU, conhecida popularmente como "pariparoba" e do princípio ativo, 4-NC, mostraram que o extrato bruto de raiz é mais ativo do que o 4-NC isolado como inibidor de metaloproteinases (MMPs) e como antioxidante, sugerindo a presença de outros compostos com estas atividades, além de um possível efeito sinérgico. O objetivo deste estudo foi o de obter frações do extrato de PU isentas de 4-NC e avaliar a atividade antioxidante, fotoprotetora e inibitória de metaloproteinases 2 e 9 na pele. O extrato hidroalcoólico de raiz de PU foi submetido ao fracionamento líquido/líquido sucessivamente com hexano e n-butanol, restando um resíduo aquoso. As frações n-butanólica (NBUT) e aquosa (AQ) mostraram-se isentas de 4-NC. A avaliação da atividade antioxidante por DPPH e ORAC das 3 frações obtidas mostrou maior capacidade antioxidante para a fração hexânica (HEX), seguido das frações NBUT e AQ. O mesmo perfil foi observado quanto à presença de compostos fenólicos. Entretanto não foi detectada a presença de flavonóides nas frações. O tratamento tópico com formulação gel-creme contendo o extrato de PU, a fração NBUT e a fração NBUT+4-NC, 2h antes da exposição aguda à radiação UVB (2 MED - 204.36 mJ/cm2), mostrou-se insuficiente para repor os níveis basais do sistema antioxidante da pele depletados em aproximadamente 20% pela radiação após 12h quando comparado ao controle. O mesmo tratamento não foi capaz de reduzir a formação de dímeros de pirimidina (CPD) após 2h. A avaliação após 12h mostrou redução da marcação...
Ultraviolet radiation exposure induces the development of undesirable effects, including photoageing and photocarcinogenesis. One way of preventing these effects is the use of natural antioxidants. The Pothomorphe umbellata (PU) root extract shows antioxidant and photoprotective activities, which have been attributed, in part, to 4-nerolidylcatechol (4-NC), the main compound from this plant species. In previous studies, the plant extract was more efficient than 4-NC as an antioxidant and in metalloproteinases (MMPs) inhibitory activities, suggesting the presence of others compounds that could be related with these properties, as well as a possible synergic effect. The purpose of this study was to obtain fractions from PU without 4-NC, and to evaluate the antioxidant, photoprotection and skin MMPs 2 and 9 inhibitory activity of this fractionation. The P.umbellata hidroalcoholic root extract was submitted to liquid/liquid extraction with hexane followed by n-buthanol, from which derived a final residue of aqueous extract. The n-buthanolic (NBUT) and aqueous (AQ) fractions did not show the presence of 4-NC. The evaluation of the antioxidant activity by DPPH and ORAC for the 3 fractions showed higher antioxidant activity for the hexanic (HEX) fraction, followed by the NBUT and AQ fractions. The same profile was observed for phenolic compound concentration. However the presence of flavonols in the fractions and PU root extract was not detected. The gel-cream topic treatment containing PU root extract, NBUT fraction and NBUT+4NC, 2h before acute UVB exposure (2 MED - 204,36 mJ/cm2), was insufficient to promote the normal conditions of basal level antioxidant system depleted approximately 20% after 12h of the exposure compared with the control. The same treatment evaluated by immunohistochemistry, was not able to reduce the CPD (cyclobutane pyrimidine dimers) 2h after the exposure. In 12h reduction of SBC (sunburn cells) and PCNA (proliferating cell nuclear antigen...
Assuntos
Antioxidantes/análise , Extratos Vegetais/química , Metaloproteinase 9 da Matriz , Raízes de Plantas , Pele , Piperaceae , Preparações de Plantas/uso terapêuticoRESUMO
A Pothomorphe umbellata (Piperaceae), é uma espécie medicinal nativa do Brasil, utilizada na indústria de cosméticos e protetores de pele contra raios UVA e UVB. Com o intuito de gerar informações aplicadas à propagação da espécie, o presente trabalho relacionou a coloração e a massa de sementes de P. umbellata a seu comportamento germinativo. A coloração e a massa de sementes de P. umbellata foram características adequadas para avaliar a homogeneidade fisiológica, o vigor, o potencial e o comportamento germinativo. Assim, conclui-se que, embora de germinação lenta, as sementes de coloração preta e mais densa devem ser as escolhidas quando de coleta ou de processo seletivo.
Pothomorphe umbellata (Piperaceae) is a medicinal species native to Brazil, which has been used in the cosmetic industry and in products that protect the skin against UVA and UVB rays. To generate information applied to the species propagation, the present work related the coloration and the mass of P. umbellata seeds to their germinative behavior. The coloration and the mass of seeds of P. umbellata were characteristic appropriate to evaluate physiologic homogeneity, vigor, potential and germinative behavior. Thus, although of slow germination, seeds of black and denser coloration should be chosen during collection or selective process.
Assuntos
Pigmentação/genética , Piperaceae/embriologia , Sementes/anatomia & histologia , Plantas Medicinais/embriologiaRESUMO
A maioria das pesquisas realizadas até hoje sobre os efeitos benéficos da pariparoba (Pothomorphe umbellata L. Miq) foi feita com o extrato da raíz desta espécie, e o seu emprego em larga escala comprometeria a exploração sustentável deste insumo natural. Neste sentido o uso das folhas da pariparoba, ao invés de raízes pela indústria cosmética, não põe em risco a existência da espécie. Neste trabalho foi determinada a concentração de 4-nerolidilcatecol (4-NC) de um extrato de folhas por metodologia analítica validada em nosso laboratório, cujo valor foi cerca de 30 por cento menor que no extrato de raiz, obtido da mesma maneira. Na avaliação de fotoestabilidade do extrato de folhas, uma solução de 0,25 mg/mL não apresentou alterações significativas do perfil espectroscópico após 2 horas de exposição à radiação UVB, demonstrando sua estabilidade. Metaloproteinases (MMPs) são endopeptidases dependentes de zinco, envolvidas na remodelagem da matriz extracelular (MEC), e importantes na formação das rugas típicas do fotoenvelhecimento cutâneo. Também avaliamos, por técnica de zimografia, a capacidade do extrato de folhas de P. umbellata de inibir a atividade de MMPs 2 e 9 in vitro de pele de camundongos sem pêlo. O extrato de folhas (0,1 mg/mL) inibiu em 80 por cento a atividade destas enzimas, conforme avaliação densitométrica.
Most researches that have been done until today about the beneficial effects of pariparoba (Pothomorphe umbellata L. Miq) have been done with root extract of this species, but the use in large scale would compromise the sustainable exploration of this natural resource. In this sense, the utilization of pariparoba leaves, substituting the roots, in the cosmetic industry does not put in risk the existence of the species. In this work the concentration of 4-nerolidylcathecol (4-NC) in leaf extract was determined by the analytical methodology validated in our laboratory. The concentration of 4-NC in leaf extract was around 30 percent less than that of root extract, obtained in the same way. Concerning the study of the photostability of a leaves extract solution containing 4-NC did not demonstrate meaningful alterations in the spectrometry profile after 2 hours of exposure under UVB radiation, showing its stability under this conditions. Metalloproteinases (MMPs) are endopeptidases that are zinc-dependent, involved in remodeling extracellular matrix (ECM), that are important in the appearance of typical photoaging wrinkles. In this work the capacity of leaf extract of P. umbellata to inhibit MMP-2 and 9 activities of hairless mouse skin in vitro by zymography gel was also evaluated. The leaf extract (0,1 mg/mL) inhibit in 80 percent activity of this enzymes, according to the densitometric zymography evaluation.
Assuntos
Animais , Camundongos , Metaloproteases , Neoplasias Cutâneas/tratamento farmacológico , Fitoterapia , Extratos Vegetais , Neoplasias Induzidas por RadiaçãoRESUMO
A atividade biológica do extrato bruto e as várias frações obtidas de folhas de Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck foi avaliada em Leishmania amazonensis. Este estudo incluiu o processo de extração e o fracionamento biomonitorado pelo método de cromatografia de adsorção. Um aumento progressivo no efeito antileishmania foi observado durante o processo de purificação. O extrato hidroalcoólico solúvel em água (EBA) apresentou concentração inibitória 50 por cento (IC50) igual a 167 µg/mL enquanto o extrato hidroalcoólico solúvel em acetato de etila (EBAcOEt) mostrou um IC50 de 30 mg/mL sobre o crescimento de formas promastigotas após 48 h de cultivo. A fração hexano (FHex) apresentou uma atividade antileishmania maior que o EBAcOEt com IC50 de 21,5 µg/mL. Análises de citotoxicidade indicaram que as concentrações tóxicas do EBA, EBAcOEt e das frações foram maiores para macrófagos J774G8 do que para os protozoários.
Biological activity of the crude extract and several fractions obtained from Piper regnellii var. pallescens was assessed on Leishmania amazonensis. This study included the extraction process and bioassay-guided fractionation by the adsorption chromatography method. A progressive increase in the antileishmanial effect was observed in the course of the purification process. The hydroalcoholic extract water soluble (EBA) had a 50 percent inhibitory concentration (IC50) at 167 µg/mL whereas the hydroalcoholic extract acetate soluble (EBAcOEt) showed an IC50 of 30 µg/mL against the growth of promastigote forms after 48 h of culturing. The hexan fraction (FHex) showed an antileishmanial activity greater than EBAcOEt with IC50 at 21.5 µg/mL. Analysis of cytotoxicity indicated that the toxic concentrations of the EBA, EBAcOEt, and fractions were higher for J774G8 macrophages than for the protozoans.