Phyllanthus emblica fruit extract alleviates halitosis and reduces the inflammatory response to oral bacteria

Abstract Objective To assess the efficacy of Phyllanthus emblica extract in alleviating halitosis and reducing the inflammatory response to halitosis-related bacteria. Methodology This investigation, using Phyllanthus emblica fruit extract (PE), involved four aspects. First, we evaluated the effect on growth and aggregation of halitosis-related bacteria, including Fusobacterium nucleatum, Porphyromonas gingivalis, and Solobacterium moorei, using a microdilution assay and scanning electron microscopy. Second, volatile sulfur compound (VSC) levels were measured on individuals with halitosis in randomized short-term (26 participants) and double-blind randomized long-term trials (18 participants in each group) after rinsing with PE for 3, 6, and 12 h, and 28 days. Third, we analyzed pro-inflammatory cytokine expression in TR146 cells using quantitative real-time PCR and enzyme-linked immunosorbent assays. Lastly, we assessed pro-inflammatory cytokine secretion and Toll-like receptor (TLR) 2 mRNA expression via the same experimental methods in a three-dimensional oral mucosal epithelial model (3D OMEM). Results PE extract dose-dependently inhibited the growth of F. nucleatum (50% inhibition concentration [IC50]=0.079%), P. gingivalis (IC50=0.65%), and S. moorei (IC50=0.07%) and effectively prevented bacterial aggregation. Furthermore, VSC contents decreased significantly at 3, 6, and 12 h after rinsing with 5% PE compared with those in the control. Long-term use of mouthwash containing 5% PE for 28 days led to a significant decrease in VSC contents. PE attenuated the F. nucleatum- or P. gingivalis-stimulated mRNA expression and protein release of interleukin (IL)-6 and IL-8 in TR146 cells. It also suppressed IL-8 and prostaglandin E2 secretion and TLR2 mRNA expression in F. nucleatum-induced OMEMs. Conclusion Our findings support the use of PE in oral care products to alleviate halitosis and it may reduce inflammation.

Polyphenolic compounds of Phyllanthus amarus Schum & Thonn. (1827) and diabetes-related activity of an aqueous extract as affected by in vitro gastrointestinal digestion.

ETHNOPHARMACOLOGICAL RELEVANCE: Extracts of the aerial part of Phyllanthus amarus have been extensively used in several countries to cure diabetes. No data is available on the impact of gastrointestinal digestion of such crude extracts on their antidiabetic activity. AIM OF THE STUDY: The aim of this study was to identify active fractions and compounds of fresh aerial parts of P. amarus extracted by an infusion method that are responsible for antidiabetic effects occurring at the level of glucose homeostasis. MATERIALS AND METHODS: An aqueous extract was obtained by an infusion method and its polyphenolic composition was analysed by reverse phase UPLC-DAD-MS. The influence of in vitro gastrointestinal digestion was evaluated both on the chemical composition and on the antidiabetic effect of P. amarus infusion extract using glucose-6-phosphatase enzyme inhibition and stimulation of glucose uptake. RESULTS: Analysis of the chemical composition of the crude extract revealed the presence of polysaccharides and various families of polyphenols such as phenolic acids, tannins, flavonoids and lignans. After simulated digestion, the total content of polyphenols decreased by about 95%. Caffeoylglucaric acid derivates and lignans exhibited strong stimulation of glucose uptake similar to metformin with an increase of 35.62 ± 6.14% and 34.74 ± 5.33% respectively. Moreover, corilagin, geraniin, the enriched polysaccharides fraction and the bioaccessible fraction showed strong anti-hyperglycemic activity with about 39-62% of glucose-6-phosphatase inhibition. CONCLUSION: Caffeoylglucaric acid isomers, tannin acalyphidin M1 and lignan demethyleneniranthin were reported for the first time in the species. After in vitro gastroinstestinal digestion, the composition of the extract changed. The dialyzed fraction showed strong glucose-6-phosphatase inhibition.

Evaluation of Salicylic Acid and Methyl Jasmonate as Elicitors in Phyllanthus acuminatus Hairy Roots by Non-Targeted Analysis Using High-Resolution Mass Spectrometry.

Phyllanthus acuminatus has been studied for its vast medical and industrial potential. Phytochemical investigations reveal that the genus is a rich source of lignans, flavonoids, phenolics, terpenoids, and other metabolites. However, the phytochemical profile elucidation of this species still needs further research. The use of eliciting compounds such as salicylic acid and methyl jasmonate has managed to increase the production of secondary metabolites in plant cell cultures. Hairy roots of Phyllanthus acuminatus were produced in 250 mL flasks with a 16 h light/8 h darkness photoperiod under diffused light with a culture time of four weeks. The elicitors salicylic acid and methyl jasmonate were tested in 50 µM and 200 µM concentrations. Non-targeted analysis was done for the different treatments using HR-MS. Identified metabolites were grouped in phenylpropanoids, phenols, and mucic acids, and statistical analysis of relative concentrations was achieved. A significant change in phenols' relative concentrations appeared in the elicitations with salicylic acid. Because of the elicitation treatment, specific compounds increased their concentrations, some of which have known pharmacological effects and are used in treating chronic diseases. The best elicitation treatment was salicylic acid 50 µM as it increased by more than 100% the general content of phenols and phenylpropanoid derivates and triplicates the concentration of mucic acid derivates in treated hairy root extracts. The application of non-targeted analysis showed interesting changes in phytochemical concentration due to elicitation in Phyllanthus acuminatus hairy roots.

Phyllanthaceae in the Atlantic Forest of northeastern Brazil / Phyllanthaceae na Mata Atlântica do Nordeste brasileiro

Abstract: The taxonomy of Phyllanthaceae Martinov in the Atlantic Forest of northeastern Brazil was updated through the analysis of approximately 200 specimens deposited in regional herbaria as well as field observations. Thirty-five species were recorded, belonging to seven genera: Amanoa Aubl. (1 species), Astrocasia B.L. Rob. & Millsp. (1), Discocarpus Klotzsch (1), Hieronyma Allemão (2), Margaritaria L.f. (1), Phyllanthus L. (28), and Richeria Vahl (1). Of the 35, six are new records for Alagoas State, two for Rio Grande do Norte, four for Paraíba, and six for Sergipe. Among the recorded species, 18 are endemic to Brazil, and of those, 11 are endemic to the Brazilian northeast and nine are exclusive to the Atlantic Forest. An identification key, comments on their taxonomy, phenology and geographic distributions, species conservation status, distribution maps, and illustrations of the species are provided.

Resumo: Este estudo teve como objetivo atualizar a taxonomia de Phyllanthaceae Martinov na Mata Atlântica do Nordeste brasileiro, através da análise de aproximadamente 200 espécimes depositados nos herbários da região e observação das espécies em campo. Foram registradas 35 espécies, pertencentes a sete gêneros: Amanoa Aubl. (1 espécie), Astrocasia B.L. Rob. & Millsp. (1), Discocarpus Klotzsch (1), Hieronyma Allemão (2), Margaritaria L.f. (1), Phyllanthus L. (29), e Richeria Vahl (1). Das 35 espécies, seis são novos registros para Alagoas, dois para o Rio Grande do Norte, quatro para Paraíba e seis para Sergipe. Dentre as espécies registradas, 18 são endêmicas do Brasil, destas, onze são endêmicas do Nordeste e nove são exclusivas da Mata Atlântica. São fornecidos chave de identificação, comentários taxonômicos, fenológicos e de distribuição geográfica, bem como o status de conservação das espécies. Além disso, são apresentados mapas de distribuição e ilustrações das espécies.

Actividad citotóxica y antiproliferativa in vitro del extracto acuoso de Phyllanthus comosus

RESUMEN Introducción: Desde los inicios de la medicina antigua, las plantas han sido utilizadas como tratamiento en diversas enfermedades incluyendo las de naturaleza infecto-contagiosa y el cáncer. Son numerosos los informes sobre las propiedades biológicas del género Phyllanthus. Objetivo: Evaluar la actividad citotóxica y antiproliferativa de un extracto acuoso de Phyllanthus comosus en tres líneas celulares, dos de origen tumoral (SiHa y HeLa) y una no tumoral (Vero). Métodos: La actividad citotóxica se evaluó mediante el método del MTT y la capacidad antiproliferativa mediante el ensayo de detección de inhibición de colonias o clonogénico. Se tuvieron en cuenta valores como la concentración citotóxica media (CC50), índice selectivo y porcentaje de disminución de la proliferación celular. Resultados: En el ensayo de citotoxicidad se obtuvieron CC50 similares para ambas líneas tumorales; mientras que el valor para la línea Vero resultó tres veces menos tóxico, con valores de índice de selectividad mayor que tres. El ensayo clonogénico demostró inhibición de la proliferación en las líneas tumorales, mientras que en células Vero no se observó inhibición de la capacidad de formación de colonias. Conclusiones: El extracto de P. comosus es más citotóxico para las líneas tumorales SiHa y HeLa que para las células Vero, no tumorales. Además, la inhibición de la formación de clonos celulares en ambas líneas tumorales evidencia su acción antiproliferativa y selectiva, lo que argumenta su potencialidad antitumoral in vitro

ABSTRACT Introduction: Ever since the onset of ancient medical practice, plants have been used to treat a variety of conditions, including infectious communicable diseases and cancer. A large number of reports are available about the biological properties of the genus Phyllantus. Objective: Evaluate the cytotoxic and antiproliferative activity of an aqueous extract of Phyllanthus comosus on three cell lines: two of tumoral origin (SiHa and HeLa) and one of non-tumoral origin (Vero). Methods: Cytotoxic activity was evaluated by the MTT method, and antiproliferative capacity by colony inhibition detection or clonogenic assay. Mean cytotoxic concentration (CC50), selective index and cell proliferation reduction percentage were some of the values taken into account. Results: The cytotoxicity assay obtained similar CC50 values for both tumor cell lines, whereas the value for the Vero line was three times less toxic, with a selectivity index above three. The clonogenic assay revealed proliferation inhibition in the tumor cell lines, whereas no inhibition of colony forming capacity was observed in Vero cells. Conclusions: The P. comosus extract is more cytotoxic for tumoral cell lines SiHa and HeLa than for non-tumor Vero cells. Additionally, inhibition of the formation of cell clones in both tumor cell lines is evidence of its antiproliferative and selective action, substantiating its in vitro antitumor potential.

Evaluation of prolyl oligopeptidase and acetylcholinesterase inhibition by Phyllanthus tenellus Roxb. from Brazil.

Phyllanthus tenellus Roxb. (Phyllanthaceae) is a plant used in Brazilian folk medicine for the treatment of intestinal infections and diabetes. Despite its use in traditional medicine, it was reported that P. tenellus extract may cause several effects in the central nervous system (CNS) of animals, such as agitation and signs of depression. The aim of this study was to determine the main constituents of P. tenellus methanol extract and to investigate whether the extract is able to inhibit the enzymes prolyl oligopeptidase (POP), acetylcholinesterase (AChE) and dipeptidyl peptidase-IV (DPP-IV). Corilagin (1) was isolated as the main constituent of the P. tenellus extract, along with rutin (2) and vitexin-2″-O-rhamnoside (3). The extract presented the ability to inhibit mainly POP. Dichloromethane and ethyl acetate fractions showed the highest inhibitory potency against POP (IC50 values of 1.7 ± 0.4 and 11.7 ± 2 µg/mL, respectively). All fractions were inactive against AChE. Corilagin displayed selective POP inhibition in a dose-dependent manner, with IC50= 19.7 ± 2.6 µg/mL. Corilagin exhibited moderate capacity to pass through the phospholipid membrane by passive diffusion, presenting effective permeability (Pe) of 1.26 × 10-7 cm/s.

In Vitro Calli Production Resulted in Different Profiles of Plant-Derived Medicinal Compounds in Phyllanthus amarus.

The efficient production of plant-derived medicinal compounds (PDMCs) from in vitro plants requires improvements in knowledge about control of plant or organ development and factors affecting the biosynthesis pathway of specific PDMCs under in vitro conditions, leading to a realistic large-scale tool for in vitro secondary metabolite production. Thus, this study aimed to develop an in vitro technique, through the induction and proliferation of calli, for production of plant fresh weight, and to compare the PDMC profile obtained from the plants versus in vitro calli of Phyllanthus amarus. It was successfully possible to obtain and proliferate two types of calli, one with a beige color and a friable appearance, obtained in the dark using Murashige and Skoog (MS) medium plus 2,4-dichlorophenoxyacetic acid (2,4-D), and a second type with a green color, rigid consistency, and nonfriable appearance obtained under light conditions and MS medium plus 6-benzyladenine (6-BA). In vitro micropropagated plants that gave rise to calli were also acclimatized in a greenhouse and cultivated until obtaining the mass for PDMC analysis and used as a control. While the micropropagated-derived plants concentrated the lignans niranthin, nirtetralin, and phyllanthin, the Phyllanthus amarus calli proliferated in vitro concentrated a completely different biochemical profile and synthesis of compounds, such as betulone, squalene, stigmasterol, and ß-sitosterol, in addition to others not identified by GC-MS database. These results demonstrate the possibility of applying the calli in vitro from Phyllanthus amarus for production of important PDMCs unlike those obtained in cultures of differentiated tissues from field plants.

Otimização do processo extrativo de Phyllanthus niruri L / Optimization of the extraction process of Phyllanthus niruri L

Phyllanthus niruri L., conhecida como quebra-pedra, é uma planta da família Phyllanthaceae, pertencente ao gênero Phyllanthus. Neste estudo foi avaliado o planejamento fatorial para estudar a importância dos fatores (concentração de etanol e processo extrativo) na concentração de flavonoides e compostos fenólicos presentes nos extratos de P. niruri. Diferentes extratos de P. niruri foram preparados utilizando etanol 50 e 96%, empregando três métodos extrativos diferentes: maceração, percolação e ultrassom. A quantificação de flavonoides foi realizada pelo método espectrofotométrico com AlCl3 10% (v/v) em água. Já o método utilizado para a quantificação de compostos fenólicos foi de Folin-Ciocalteu. As análises foram feitas em triplicata e os resultados analisados nos softwares Minitab18® e Prisma GraphPad®. Foram observadas diferenças significativas (p≤0,05) nos valores de flavonoides e compostos fenólicos totais nos extratos produzidos de P. niruri, quando foram utilizados diferentes processos extrativos, bem como diferentes concentrações de etanol. Analisando os gráficos de Pareto, foi possível verificar que a concentração de solvente foi o efeito que mais contribuiu na extração de ambos os compostos bioativos. Verificando os gráficos de interações, foi possível identificar que o etanol 96% influenciou positivamente na extração de flavonoides e o etanol 50% na extração de compostos fenólicos. Assim, o planejamento fatorial aplicado ao processo extrativo de P. nirurievidenciou que a extração de flavonoides pode ser realizada com eficiência por métodos extrativos tradicionais, maceração ou percolação, utilizando como solvente o etanol 96%. Já as condições otimizadas para a extração de compostos fenólicos são por percolação ou ultrassom com etanol 50%.

Phyllanthus niruri L., known as stone breaker, is a plant in the Phyllanthaceae family, belonging to the genus Phyllanthus. In this study, the factorial design was evaluated to study the importance of factors (concentration of ethanol and extraction process) on the concentration of flavonoids and phenolic compounds present in P. niruri extracts. Different extracts of P. niruri was prepared using 50 and 96% ethanol, using three different extraction methods: maceration, percolation and ultrasound. The quantification of flavonoids was performed by the spectrophotometric method with 10% AlCl3 (v/v) in water. The Folin-Ciocalteu method were employed for the quantification of phenolic compounds. The analyses were performed in triplicate and the results were analyzed using the Minitab18® and Prisma GraphPad® software. Significant differences (p≤0.05) were observed in the values of flavonoids and total phenolic compounds in the extracts produced from P. niruri, when different extraction processes were used, as well as different concentrations of ethanol. Analyzing the Pareto graphs, it was possible to verify that the solvent concentration was the effect that most contributed to the extraction of both bioactive compounds. Checking the interaction graphs, it was possible to identify that 96% ethanol positively influenced the extraction of flavonoids and 50% ethanol influenced the extraction of phenolic compounds. Thus, the factorial design applied to the extraction process of P. niruri showed that the extraction of flavonoids can be carried out efficiently by traditional extraction methods, maceration or percolation, using 96% ethanol as a solvent. The optimized conditions for the extraction of phenolic compounds are by percolation or ultrasound with 50% ethanol.

Insights into the Phytochemistry of the Cuban Endemic Medicinal Plant Phyllanthus orbicularis: Fideloside, a Novel Bioactive 8-C-glycosyl 2,3-Dihydroflavonol.

Phyllanthus orbicularis (Phyllanthaceae) is an endemic evergreen tropical plant of Cuba that grows in the western part of the island and is used in traditional medicine as an infusion. The aqueous extract of this plant presents a wide range of pharmacological activitiessuch as antimutagenic, antioxidant and antiviral effects. Given the many beneficial effects and the great interest in the development of new pharmacological products from natural sources, the aim of this work was to investigate the phytochemistry of this species and to elucidate the structure of the main bioactive principles. Besides the presence of several known polyphenols, the major constituent was hitherto not described. The chemical structure of this compound, here named Fideloside, was elucidated by means of HR-ESIMS/MSn, 1D/2D NMR, FT-IR, and ECD as (2R,3R)-(-)-3',4',5,7-tetrahydroxydihydroflavonol-8-C-ß-D-glucopyranoside. The compound, as well as the plant aqueous preparations, showed promising bioactive properties, i.e., anti-inflammatory capacity in human explanted monocytes, corroborating future pharmacological use for this new natural C-glycosyl flavanonol.

UPLC-HRMS and NMR applied in the evaluation of solid-phase extraction methods as a rational strategy of dereplication of Phyllanthus spp. aiming at the discovery of cytotoxic metabolites.

The classical approach to drug discovery from natural products (NP's) requires strenuous and complex purification steps for the isolation and structural elucidation. Modern strategies as dereplication aim to accelerate the identification of known compounds present in a crude or partially purified extract. In this work, we investigated the influence of the solid-phase extraction (Oasis, Plexa, and Agilent C18 cartridges with and without organic modifiers) chemical profile obtained by UPLC-QTOF-MS and NMR and cytotoxicities of aqueous extracts from Phyllanthus niruri and P. amarus. Our results showed differences between the SPE cartridges and the mass recovered. P. niruri showed higher mass recovery than P. amarus indicating a higher amount of secondary metabolites. The UPLC-QTOF-MS analysis revealed that P. niruri crude extract presents higher contents of phenolic compounds than P. amarus. According to NMR analysis, P. niruri contained more tyrosine, corilagin, and glycosidic residues while P. amarus, presented higher content of ellagic acid. The different stationary phases, as well as mobile phases for exploratory SPE, enabled the exploitation of the different chemical functionalities within the Phyllanthus species. The SPE (MeOH:H2O 70:30 with C18 cartridges) samples showed greater in vitro cytotoxicity than the crude extracts, with IC50 ranging from 8.01 to 94.92 µg mL-1 against the tumor lines tested. The solid phase extraction allowed the concentration of molecules with desirable physicochemical characteristics, which might increase the hit of therapeutically useful substances.

Antimicrobial activity of Phyllanthus amarus Schumach. & Thonn and inhibition of the NorA efflux pump of Staphylococcus aureus by Phyllanthin.

The aim of this study was to evaluate the antimicrobial activity of ethanoic extract of P. amarus (PAEE) and its compound Phyllanthin, as well as, investigate if these natural products could modulate the fluoroquinolone-resistance in S. aureus SA1199-B by way of overexpression of the NorA efflux pump. Microdilution tests were carried out to determine the minimal inhibitory concentration (MIC) of the PAEE or Phyllanthin against several bacterial and yeast strains. To evaluate if PAEE or Phyllanthin were able to act as modulators of the fluoroquinolone-resistance, MICs for Norfloxacin and ethidium bromide were determined in the presence or absence of PAEE or Phyllanthin against S. aureus SA1199-B. PAEE showed antimicrobial activity against Gram-negative strains, meanwhile Phyllanthin was inactive against all strains tested. Addition of PAEE or Phyllanthin, to the growth media at sub-inhibitory concentrations enhanced the activity of the Norfloxacin as well as, Ethidium Bromide, against S. aureus SA1199-B. These results indicate that Phyllanthin is able to modulate the fluoroquinolone-resistance possibly by inhibition of NorA. This hypothesis was supported by in silico docking analysis which confirmed that Phyllantin is a NorA ligand. Thus, this compound could be used as a potentiating agent of the Norfloxacin activity in the treatment of infections caused by fluoroquinolone-resistant S. aureus.

Acetylcholinesterase inhibition, antioxidant and identification of some chemical constituents of Phyllanthus atropurpureus cultivated in Egypt

Investigation of the lipid constituents of the aerial parts of Phyllanthus atropurpureus resulted in isolation and identification of the fatty acid mixture which consists of eight acids with linolenic acid as major and the unsaponifiable fraction that contain a series of hydrocabons, sterols, in addition to one triterpene (α-amyrin). The acetone insoluble fraction was found to contain two fatty alcohols and three n-hydrocabons in which the n-eicosane is the most abundant (44.16%). The flavonoidal constituents were isolated from ethyl acetate and butanol fractions which were identified as: luteolin-7-O-glucoside, kaempferol 3-O-(p-coumaroylglucoside), kaempferitrin, luteolin and kaempferol. Evaluation of different extracts as acetylcholinesterase inhibitors (AChI), established the chloroform fraction as a promising inhibitor of the enzyme. The antioxidant testing with DPPH radical revealed the potential of precipitate from MeOH extract as a radical scavenger.

HPLC fingerprint and simultaneous quantitative analysis of phyllanthin and hypophyllanthin for identification and authentication of Phyllanthus niruri from related species

Abstract A precise and accurate method for the identification and authentication of Phyllanthus niruri L. from P. debilis Klein ex Willd. and P. urinaria L., Phyllanthaceae, was developed using high-performance liquid chromatography. Chromatographic fingerprint analysis was combined with simultaneous quantification of phyllanthin and hypophyllanthin for the developed method. Phyllanthin and hypophyllanthin were successfully separated and quantified under this proposed method. The highest amount of phyllanthin and hypophyllanthin was found in P. niruri compared to P. debilis and P. urinaria. Fingerprint chromatogram of the three Phyllanthus species showed distinct profiles that these may be used to identify and authenticate each Phyllanthus species, which improved by marker compounds present in each species. The combination of chromatographic fingerprint analysis and discriminant analysis was successfully discriminated all three species, including P. niruri adulterated with P. debilis or P. urinaria. The method can be used for the identification and authentication of P. niruri from related species, such as P. debilis and P. urinaria.

Anti-Inflammatory and Antinociceptive Studies of Hydroalcoholic Extract from the Leaves of Phyllanthus brasiliensis (Aubl.) Poir. and Isolation of 5-O-ß-d-Glucopyranosyljusticidin B and Six Other Lignans.

The aim of this study was to investigate the chemical composition and the antiinflammatory/antinociceptive properties of the hydroalcoholic extract derived from the leaves of Phyllanthus brasiliensis (HEPB) in rodents. A new arylnaphthalene lignan glycoside, 5-O-ß-d-glucopyranosyljusticidin B, together with six known lignans, were isolated from HEPB. 1D and 2D NMR experiments and HRMS were used to elucidate the structure of the new compound. HEPB toxicity and antinociceptive activity were evaluated through acute oral toxicity and formalin models in mice, respectively. The anti-inflammatory effects of HEPB were assessed using carrageenan- and dextran-induced paw edema models in rats. HEPB showed low toxicity. Oral administration of HEPB reduced paw edema induced by carrageenan, but not by dextran. HEPB and its fractions from FR6 to FR10 (FR6-10) inhibited the neurogenic and inflammatory phases of formalin-induced linking, demonstrating its antinociceptive activity. These results indicated that lignans from Phyllanthus brasiliensis exerted antinociceptive/anti-inflammatory effects not related to the histaminergic pathway.

Cuban flora species as a potential source of DNA protective compounds / Especies de la flora cubana cómo fuente potencial de compuestos protectores del DNA

Environmental exposure to genotoxic agents represents a major health concern for modern society. DNA damage could lead to mutations, which accumulative effect is closely related to degenerative and lethal diseases, such as cancer. Because of their structural and chemical diversity natural products play a fundamental role in pharmaceutical sciences for novel drug discovery. The present review article focuses on pre-clinical studies done with some species from Cuban flora that have been tested with positive antigenotoxic properties against different genotoxins. Special emphasis regarding molecular mechanisms suggested, from antioxidant activity to DNA repair modulation, a critical discussion of the state of art and the perspectives in the use of these plants as a new and promising strategy for genoprotection in the 21st Century are included.

La exposición ambiental a agentes genotóxicos representa un problema de salud significativo en la sociedad actual. El daño al ADN puede generar mutaciones, cuyo efecto acumulativo se encuentra estrechamente relacionado con enfermedades degenerativas y letales como el cáncer. Debido a su diversidad estructural y química los productos naturales juegan un papel fundamental en las ciencias farmacéuticas en el descubrimiento de nuevas drogas. El presente artículo de revisión puntualiza estudios pre-clínicos realizados con determinadas especies de la flora cubana que han sido estudiadas con una respuesta antioxidante positiva frente a diferentes genotoxinas. Se enfatizan especialmente los mecanismos moleculares sugeridos, desde actividad antioxidante hasta modulación de la reparación del ADN, así como una discusión crítica del estado del arte y las perspectivas en el empleo de estas plantas como una estrategia nueva y prometedora para la genoprotección en el siglo 21.

Schistosoma mansoni: In vivo evaluation of Phyllanthus amarus hexanic and ethanolic extracts.

Human schistosomiasis is an important neglected tropical disease caused by blood flukes of the genus Schistosoma and is responsible for more than 280,000 deaths annually. Treatment for this disease relies currently on a single drug, praziquantel (PZQ). Concerns regarding PZQ resistance and insensitivity of juvenile schistosomes have increased the interest in resorting to medicinal plants for alternative drug therapies. This study aimed to perform an in vivo schistosomicidal activity evaluation of crude hexanic (HE) and ethanolic (EE) extracts obtained from Phyllanthus amarus in mice infected with Schistosoma mansoni (BH strain). Mice were treated orally with a single dose of 100 or 250 mg/kg, on two different infection periods, 30 and 45 days post-infection (dpi). Parameters such as worm recovery, faecal egg count, intestinal tissue egg count and liver histopathology were evaluated. Treatment against young adult (30 dpi) and adult (45 dpi) worms were more effective compared to the control group treated with PZQ. At a concentration of 250 mg/kg (30 dpi) EE showed a 54.4% female reduction and a 61.2% total worm reduction whilst at a concentration of 100 mg/kg (45 dpi) HE showed a 40.6% female worm reduction and a 45.3% total worm reduction. Histopathological examination showed a granuloma decrease in both number and size for groups treated with 250 mg/kg of HE (45 dpi) or EE (30 or 45 dpi). From these results, it can be concluded that both hexanic and ethanolic extracts have antischistosomal activities, however, act differently according to the parasites age. The schistosomicidal activity results in groups treated 30 days post infection is extremely important since praziquantel does not show activity against the juvenile forms of Schistosoma.

Comparative antioxidant and bioavailability studies of Vitamin C in Phyllanthus emblica Linn. and its combinations with Piper nigrum Linn. and Zingiber officinale Roscoe

ABSTRACT Phyllanthus emblica Linn. (amla) is used in Ayurveda, the ancient Indian system of medicine and its major constituent is vitamin C which has effective free radical scavenging property. The purpose of this study was to evaluate the in vitro antioxidant activity and the bioavailability profile of vitamin C in amla and its combinations with piperine and ginger in comparison to synthetic vitamin C using New Zealand rabbits. In vitro antioxidant activity studies of synthetic vitamin C, amla, amla with piperine and amla with ginger were carried out using different models such as 2,2-Diphenyl-1-picrylhydrazyl, Nitric Oxide, Hydrogen peroxide scavenging methods, Total reductive capability and Oxygen Radical Absorbance Capacity estimation. The study results showed that synthetic vitamin C, amla, amla with piperine and amla with ginger possess significant in vitro antioxidant activity. For bioavailability studies, synthetic vitamin C, amla, amla with piperine and amla with ginger 100 mg/kg, were administered orally and the serum samples were analyzed by HPLC at 0, 1, 2, 3, 4, 6, 8, 10, 12 and 24 hours. Bioavailability studies revealed that amla with piperine combination has higher concentration of vitamin C when compared to synthetic vitamin C. This is probably due to presence of piperine, which is a bioavailability enhancer. The present study supports the fact that amla with piperine combination can be an alternative to synthetic vitamin C.

RESUMO Phyllanthus emblica Linn. (amla) é utilizada na medicina Ayurveda, medicina da Índia antiga e seu principal constituinte é a vitamina C, que possui propriedade sequestrante de radicais livres. O propósito deste estudo foi avaliar a atividade antioxidante in vitro e o perfil de biodisponibilidade da vitamina C na amla e suas combinações com piperina e gengibre em comparação com a vitamina C sintética, utilizando coelhos da Nova Zelândia. Os estudos de atividade antioxidante in vitro de vitamina C sintética, amla, amla com piperina e amla com gengibre foram realizados utilizando-se diferentes modelos para sequestrantes, como 2,2-difenil-1-picrilidrazil, óxido nítrico, peróxido de hidrogênio, capacidade redutiva total e a estimativa da capacidade de absorvância do radical oxigênio. Os resultados do estudo mostraram que vitamina C sintética, amla, amla com piperina e amla com gengibre possuem atividade antioxidante in vitro significativa. Para os estudos de biodisponibilidade, administraram-se oralmente vitamina C sintética, amla, amla com piperina e amla com gengibre 100 mg/kg e as amostras de soro foram analisadas por CLAE em 0, 1, 2, 3, 4, 6, 8, 10, 12 e 24 horas. Os estudos de biodisponibilidade revelaram que a associação de amla com piperina tem maior concentração de vitamina C, quando comparada com a vitamina C sintética. Este efeito é provavelmente devido à presença de piperina, que é intensificador de biodisponibilidade. O presente estudo apoia o fato de que a associação de amla e piperina pode ser uma alternativa para a vitamina C sintética.

Phyllanthus emblica l. branch extract ameliorates testicular damage in valproic acid-induced rats / El extracto de la rama de Phyllanthus emblica l. disminuye el daño testicular inducido por ácido valproico en ratas

Valproic acid (VPA), widely used in treating epileptic patients, can damage reproductive parameters causing male infertility. This study aimed to investigate protective effect of Phyllanthus emblica L. branch (PE) extract on rat testicular damage induced with VPA. Male rats were divided into 6 groups (control, VPA, 250 mg/kgBW PE only, and 50, 100, 250 mg/kgBW PE+VPA, respectively). Animals were pretreated with PE for 23 days and co-administered with VPA for 10 days before all reproductive parameters were determined. The results showed all doses of PE significantly protected the decrease testicular weight and testosterone level in VPA rats. PE significantly improved the decrease sperm concentration in VPA treated rats. Moreover, testicular histology of PE+VPA groups showed declining of testicular histopathologies as compared to VPA group. Therefore, it seems that PE branch extract can prevent testicular damages including male reproductive parameters in rats induced with VPA.

El ácido valproico (AVP) es utilizado frecuentemente en el tratamiento de pacientes epilépticos y puede dañar los parámetros reproductivos que causan la infertilidad masculina. Este estudio tuvo como objetivo investigar el efecto protector de la rama Phyllanthus emblica L. (PE) sobre el daño testicular de ratas inducidas con AVP. Ratas machos fueron divididas en 6 grupos (control, AVP, PE 250 mg/kg peso corporal, APV+ PE 50, 100, 250 mg/kg peso corporal, respectivamente). Los animales fueron pretratados con PE durante 23 días y se administró AVP durante 10 días antes de medir todos los parámetros reproductivos. Los resultados mostraron que todas las dosis de PE protegen significativamente el peso y los niveles reducidos de testosterona testicular en ratas con AVP. El extracto de PE mejoró significativamente la concentración de espermatozoides en ratas tratadas con AVP. Por otra parte, la histología testicular de los grupos PE+AVP mostró disminución de la histopatología testicular en comparación con el grupo tratado sólo con AVP. Por lo tanto, parece que el extracto de la rama PE puede prevenir daños testiculares incluyendo los parámetros reproductores masculinos en ratas inducidas con AVP.

Ethnopharmacological investigation of the diuretic and hemodynamic properties of native species of the Brazilian biodiversity.

ETHNOPHARMACOLOGICAL RELEVANCE: Although Echinodorus grandiflorus, Cuphea carthagenensis, and Phyllanthus tenellus infusions are used in Brazilian folk medicine due to their possible diuretic effect, none of these species was critically investigated as a diuretic drug. So, the aim of this study was to evaluate the possible acute diuretic activity of ethanol soluble fractions (ES) obtained from these species and assess the relationship between renal cortical blood flow and their antioxidant and hypotensive activity using normotensive Wistar rats. MATERIAL AND METHODS: The preparation obtained from E. grandiflorus (ES-EG), C. carthagenensis (ES-CC), and P. tenellus (ES-PT) infusions was orally administered in a single dose to rats. Urine excretion rate, pH, density, conductivity and Na(+), K(+), Cl(-) and HCO3(-) contents were measured in the urine of saline-loaded animals. Concentration of electrolytes, total protein, urea, creatinine, and angiotensin converting enzyme (ACE) activity were evaluated in collected serum. The involvement of the renal cortical blood flow and antioxidative activity in the hypotensive and diuretic effects was also determined. RESULTS: Water and Na(+), Cl(-) and Na(+) excretion rates were significantly increased by ES-EG, while urinary bicarbonate excretion was reduced. Moreover, ES obtained from E. grandiflorus was able to significantly increase renal blood flow and reduce mean arterial pressure and oxidative stress in "in vitro" and "in vivo" models. All other parameters evaluated were not affected by any treatment. CONCLUSION: The results presented here shown that the ES-EG obtained from E. grandiflorus leaves shown a significant diuretic and hypotensive activity and suggest that these effects could be related with an important renal and systemic vasodilator effect. In addition, it was shown for the first time that the pharmacological effects of ES obtained from P. tenellus and C. carthagenensis do not support its popular use as a diuretic agent.

Ensayo a ciego simple del efecto hipoglucemiante de phyllanthus niruri (huevito escondido) y su combinación con azadirachta indica (NIM) en ratas wistar / A simple blind test of hypoglycemic effect of phyllanthus niruri (huevito escondido) and its combination with azadirachta indica (NIM) in wistar rats

Objetivo: valorar el efecto hipoglucemiante de un extracto acuoso de la planta entera Phyllantus niruri, hojas de Azadirachta indica y su combinación. Métodos: Se diseñó un ensayo cruzado a ciego simple, preparándose extractos acuosos de hojas de A. indica y planta entera de P. niruri, colocando 70 g en 200 mL de agua a 55-60 °C, conformando tres tratamientos: uno para cada extracto y el tercero la combinación. Como control positivo o de referencia se utilizó el extracto de A. Indica. Se emplearon 18 ratas Wistar a las que se administró el tratamiento vía oral una sola vez, a razón de 5 mL/300 g de peso. La glucemia se midió con un glucómetro One Touch Ultra antes y 2h después del tratamiento. Se aplicó la prueba ANOVA a los datos. Resultados: Hubo disminución de la glucemia con los tres tratamientos. No se encontró diferencia entre los tratamientos probados. Conclusiones: el extracto acuoso de la planta entera de Phyllanthus niruri tiene un efecto hipoglucemiante similar al extracto acuoso de hojas de Azadirachta indica. No hubo evidencia de efecto sinérgico.

Objective: to evaluate the hypoglycemic effect of aqueous extract of Phyllanthus niruri whole plant and its combination with one of Azadirachta indica leaves. Methods: A single-blind crossover trial was design. One for each extract and the third corresponding to the combination: aqueous extracts of leaves of Azadirachta indica and whole plant Phyllanthus niruri, placing 70 g of the plant material in 200 mL of water at 55-60 ° C, three treatments were prepared. The comparison was performed using the extract of Azadirachta indica as control. Eighteen Wistar rats were used. Treatment was given orally at a dose of 5 mL/300 g. Glucose measurement was made with a glucometer One Touch Ultra before and 2h after administration of the treatment. ANOVA was applied to the data. Results: There was a decrease in blood glucose levels with the three treatments. No difference was found between the tested treatments regarding the hypoglycemic effect. Conclusions: The aqueous extract of the whole plant of Phyllanthus niruri has a similar hypoglycemic effect to aqueous extract of Azadirachta indica leaves. There was no evidence of synergistic effect.