Cultivo in vitro de raíces pilosas del arbusto Phyllanthus acuminatus (Phyllanthaceae) / In vitro culture of hairy roots of the shrub, Phyllanthus acuminatus (Phyllanthaceae)

Introducción: En condiciones naturales, las raíces del arbusto, Phyllanthus acuminatus, producen bajas concentraciones de metabolitos secundarios de interés medicinal. Esto abre una oportunidad para el cultivo in vitro, para aumentar la concentración de metabolitos. Objetivo: Determinar las condiciones óptimas de cultivo líquido para raíces pilosas de P. acuminatus. Métodos: Se utilizó la evaluación del crecimiento de la biomasa según porcentaje de inóculo inicial (0.50 y 0.10 %), porcentaje de nutrientes de los medios (100, 50 y 25 %) y tasa de agitación (90, 100 y 110 min-1) (N= 15 repeticiones). Resultados: Las mejores condiciones de cultivo líquido fueron: 0.10 % de inóculo inicial, nutrientes al 25 % y 90 min-1 para la tasa de agitación. Hay diferencias entre las raíces pilosas y las raíces no transformadas. Conclusiones: es factible producir raíces pilosas de P. acuminatus a gran escala, aplicando e implementando las condiciones evaluadas de porcentaje de inóculo, nutrientes en el medio y tasas de agitación utilizadas en este estudio.

Introduction: Under natural conditions, the roots of the shrub, Phyllanthus acuminatus, produce low concentrations of secondary metabolites of medicinal interest. This opens an opportunity for in vitro culture, to increase metabolite concentration. Objective: To determine the optimal liquid culture conditions for hairy roots of P. acuminatus. Methods: We used biomass growth evaluation according to initial inoculum percentage (0.50 and 0.10 %), percentage of medium nutrients (100, 50 and 25 %) and agitation rate (90, 100 and 110 min-1) (N=15 replications). Results: The best liquid culture conditions were: 0.10 % of initial inoculum, nutrients at 25 % and 90 min-1 for the agitation rate. There are differences among hairy roots and non-transformed roots. Conclusions: It is feasible to produce P. acuminatus hairy roots at a large scale, applying and implementing the evaluated conditions of inoculum percentage, nutrients in the medium and agitation rates.

Effect of phyllanthus niruri on metabolic parameters of patients with kidney stone: a perspective for disease prevention

ABSTRACT Phyllanthus niruri (P.niruri) or stone breaker is a plant commonly used to reduce stone risk, however, clinical studies on this issue are lacking. Objective: To prospectively evaluate the effect of P. niruri on the urinary metabolic parameters of patients with urinary lithiasis. Materials and Methods: We studied 56 patients with kidney stones <10mm. Clinical, metabolic, and ultrasonography assessment was conducted before (baseline) the use of P. niruri infusion for 12-weeks (P. niruri) and after a 12-week (wash out) Statistical analysis included ANOVA for repeated measures and Tukey's/McNemar's test for categorical variables. Significance was set at 5%. Results: Mean age was 44±9.2 and BMI was 27.2±4.4kg/m2. Thirty-six patients (64%) were women. There were no significant changes in all periods for anthropometric and several serum measurements, including total blood count, creatinine, uric acid, sodium, potassium, calcium, urine volume and pH; a significant increase in urinary potassium from 50.5±20.4 to 56.2±21.8 mg/24-hour (p=0.017); magnesium/creatinine ratio 58±22.5 to 69.1±28.6mg/gCr24-hour (p=0.013) and potassium/creatinine ratio 39.3±15.1 to 51.3±34.7mg/gCr24-hour (p=0.008) from baseline to wash out. The kidney stones decreased from 3.2±2 to 2.0±2per patient (p<0.001). In hyperoxaluria patients, urinary oxalate reduced from 59.0±11.7 to 28.8±16.0mg/24-hour (p=0.0002), and in hyperuricosuria there was a decrease in urinary uric acid from 0.77±0.22 to 0.54±0.07mg/24-hour (p=0.0057). Conclusions: P.niruri intake is safe and does not cause significant adverse effects on serum metabolic parameters. It increases urinary excretion of magnesium and potassium caused a significant decrease in urinary oxalate and uric acid in patients with hyperoxaluria and hyperuricosuria. The consumption of P.niruri contributed to the elimination of urinary calculi.

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.

A novel effect of geraniin on OPG/RANKL signaling in osteoblasts

In this study, the effects of geraniin on osteoprotegerin/receptor activator of nuclear factor-κB ligand(OPG/RANKL) in regulating the proliferation of osteoblasts and suppression of osteoclast-like cells (OLC) in OLC-osteoblast co-cultured system in vitro were investigated. Osteoblasts were cultured and identified with alkaline phosphatase (ALP), gomori stain, and mineralized nodule stain. OLCs were isolated from long bones of Sprague-Dawley (SD) rats and identified with tartrate-resistant acid phosphatase(TRAP) stain. Methyl thiazolyl tetrazolium assay was used to examine the proliferation of osteoblasts, and immunocytochemistry and in situ hybridization to analyze the expression OPG/RANKL in osteoblasts co-cultured with osteoclasts under the action of geraniin, respectively. Geraniin could regulate the proliferation of osteoblasts MC3T3-E1, decrease the number of OLC in OLC-osteoblast co-cultured system, and inhibit the bone resorption areas and resorption pits of OLC in vitro experiments. Geraniin could promote the mRNA and protein expression levels of OPG and suppress those of RANKL in osteoblasts. These results indicate that geraniin has a promoting effect on the proliferation of osteoblasts and an inhibitory effect on the osteoclastic bone-resorption through regulating OPG/RANKL signaling pathway in OLC-OB co-cultured system.

Liver metabolomics study reveals protective function of Phyllanthus urinaria against CCl-induced liver injury / 中国天然药物

Phyllanthus Urinaria L. (PUL) is a traditional Chinese medicine used to treat hepatic and renal disorders. However, the mechanism of its hepatoprotective action is not fully understood. In the present study, blood biochemical indexes and liver histopathological changes were used to estimate the extent of hepatic injury. GC/MS and LC/MS-based untargeted metabolomics were used in combination to characterize the potential biomarkers associated with the protective activity of PUL against CCl-induced liver injury in rats. PUL treatment could reverse the increase in ALT, AST and ALP induced by CCl and attenuate the pathological changes in rat liver. Significant changes in liver metabolic profiling were observed in PUL-treated group compared with liver injury model group. Seventeen biomarkers related to the hepatoprotective effects of PUL against CCl-induced liver injury were screened out using nonparametric test and Pearson's correlation analysis (OPLS-DA). The results suggested that the potential hepatoprotective effects of PUL in attenuating CCl-induced hepatotoxicity could be partially attributed to regulating L-carnitine, taurocholic acid, and amino acids metabolism, which may become promising targets for treatment of liver toxicity. In conclusion, this study provides new insights into the mechanism of the hepatoprotection of Phyllanthus Urinaria.

Anti-inflammatory, Anti-arthritic and Analgesic Effect of the Herbal Extract Made from Bacopa monnieriis, Cassia fistula and Phyllanthus polyphyllus

Anti-inflammatory, anti-arthritic and analgesic activity of each herbal extract, which is extracted from Bacopa monnieriis, Cassia fistula and Phyllanthus polyphyllus, respectively. The treatment of herbal extract exhibited anti-inflammatory effect as a dose-dependent manner, from 1.25mg/kg to 12.5mg/kg, in acute inflammatory models (carrageen and egg-albumin induced rat hind paw edema). It also elicited significant anti-inflammatory activity in chronic inflammatory models (cotton pellet granuloma and Freund's adjuvant induced polyarthritis in rat). In cotton pellet granuloma test, the extract exhibited the inhibitory effect of 23 and 57% at the dose of 6.25 and 12.5 mg/kg, respectively. In Freund's adjuvant induced model, the treatment of the extract of 1.25, 6.25 and 12.5 mg/kg showed the inhibitory effect of 23, 56 and 66% at 8 days, respectively. In the acetic acid-induced model, the extract significantly reduced abdominal writhing in mice when compared to the control group, reducing the mean number of writhing from 41 ± 2 in the control group to 17 ± 3 and 15 ± 2 at the dose of 6.25 and 12.5 mg/kg. From these experiments, the extract, which was extracted from the combination of Bacopa monnieriis, Cassia fistula and Phyllanthus polyphyllus, (w/w/w = 1/2/1) is surprisingly found a significant analgesic and anti-inflammatory activity.

Anatomia e histoquímica da lâmina foliar de espécies conhecidas por quebra-pedra (Euphorbia prostrata Aiton, Euphorbia hyssopifolia L., Phyllanthus amarus Schumach. & Thonn e Phyllanthus tenellus Roxb.) / Leaf anatomy and histochemistry of species known as stonebreaker (Euphorbia prostrata Aiton, Euphorbia hyssopifolia L., Phyllanthus amarus Schumach. & Thonn, and Phyllanthus tenellus Roxb.)

RESUMO O presente trabalho teve por objetivo realizar estudo anatômico comparativo entre duas espécies de Euphorbia L. e duas de Phyllanthus L. conhecidas popularmente por quebra-pedra, a fim de contribuir com a identificação das espécies, visando o controle de qualidade e testes histoquímicos para conhecimento e localização de substâncias, com intuito de fornecer subsídios para futuros estudos. As quatro espécies estudadas foram coletadas na cidade de São Mateus-ES. Foram realizadas análises anatômicas e histoquímicas da lâmina foliar. Em vista frontal, E. prostrata apresenta paredes celulares poligonais, enquanto em E. hyssopifolia as paredes são sinuosas. As espécies de Euphorbia se diferenciam pela presença de canais laticíferos e estrutura Kranz. Somente nas espécies de Phyllanthus foram observados cristais de oxalato de cálcio. P. tenellus difere de P. amarus pela presença de cristais romboédricos e P. amarus por exibir cristais prismáticos no mesofilo. O estudo anatômico foliar foi de grande relevância para a diferenciação das espécies conhecidas por quebra-pedra, revelando ser um parâmetro eficaz e que pode auxiliar no controle de qualidade de plantas medicinais, bem como os testes histoquímicos podem fornecer informações para futuros estudos com quebra-pedra.

ABSTRACT This study aimed to compare the anatomical and histochemical perspective between two species of Euphorbia L. and two of Phyllanthus L., to contribute to their identification, aiming the quality control of medicinal plans. The four studied species were collected in the city of São Mateus, ES, Brazil. We performed anatomical and histochemical analyses of leaves, and all parameters were effective for species differentiation. In the anatomical analyses, we observed crystals only in the Phyllanthus species. Euphorbia species differed by the presence of laticifer channels and Kranz structure. In frontal and cross-sectional view, P. tenellus differs from P. amarus by the presence of rhombohedral crystals. In front view, E. prostate presents polygonal cell walls, while in E. hyssopifolia the walls are winding. In the histochemical tests conducted on the leaf blade, P. amarus stands out by presenting negative reaction to ferric chloride. The anatomical and histochemical studies of leaves are really important to the knowledge and differentiation of species known as stonebreaker. However, phytochemical studies are needed for more detailed knowledge of substances from the secondary metabolism of the plant.

Extract from Phyllanthus urinaria L. inhibits hepatitis B virus replication and expression in hepatitis B virus transfection model in vitro / 中国结合医学杂志

<p><b>OBJECTIVE</b>To explore the effects of the extract from Phyllanthus urinaria L. on hepatitis B virus (HBV) replication and expression in HBV transient transfection model in vitro.</p><p><b>METHODS</b>The eukaryotic expression plasmid pHBV1.1, which contains 1.1-fold-overlength genome of HBV, was transfected into the human hepatoma cell line, HepG2, to establish and assess the HBV transient transfection model. The extract from Phyllanthus urinaria L. was prepared in different concentrations and methyl thiazolyl tetrazolium was used to detect the maximum nontoxic concentration of the drug. The extract from Phyllanthus urinaria L. were added into the transfected cell, at the concentrations of 0.8, 0.2 and 0.05 g/L, respectively. Four days after drug application, enzyme-linked immuno sorbent assay was used to detect the concentration of HBsAg in the supernatants, Southern blot was applied to analyze HBV DNA level, and Western blot was used to detect the expression of HBcAg in cells.</p><p><b>RESULTS</b>After the transfection of plasmid pHBV1.1 into HepG2 cells, the concentration of HBsAg in supernatants was increased obviously as compared with that of the normal cells (P<0.05), and all expected HBV replicative intermediates were confirmed by Southern blot analysis, which ensured the successful establishment of the HBV transient transfection model. After the application of drugs at the concentrations of 0.8 and 0.2 g/L, the level of HBsAg was obviously decreased in the supernatants, as compared with that of the virus group (P<0.05); Southern blot showed that the level of HBV rc DNA, ds DNA, ss DNA was obviously reduced compared with that of the virus group (P<0.01); Western blot revealed that the expression of HBcAg in the drug group was obviously inhibited, as compared with that of the virus group (P<0.01).</p><p><b>CONCLUSIONS</b>The extract from Phyllanthus urinaria L. obviously inhibited replication and expression of HBV in HBV transfected cell lines in vitro, thus exerting distinctive anti-HBV effects.</p>

Avaliação de extratos de quebra- pedra (Phyllanthus sp) frente à patógenos causadores de infecções no trato urinário / Evaluation of Phyllanthus sp extracts in face of Pathogen Causers of Urinary Treat Infections

RESUMO O trato urinário normalmente é estéril, no entanto, pode ser contaminado por agentes etiológicos provenientes da microbiota intestinal, dentre os mais comuns pode-se destacar a Escherichia coli. Os microrganismos estão se tornando cada vez mais resistentes a múltiplos antimicrobianos. É notória, portanto, a necessidade de encontrar novas substâncias com propriedades antimicrobianas. Portanto, foram avaliados diferentes extratos de Phyllanthus sp, frente a microrganismos causadores de infecções no trato urinário e comparadas as técnicas de hole-plate e disco difusão. Para ambas as técnicas avaliadas, o extrato de 72 horas mostrou melhor atividade antimicrobiana, na técnica de disco difusão, a bactéria mais sensível foi o Staphylococcus saprophyticcus, que apresentou CMI (Concentração Mínima Inibitória) de 15,84 mg/mL. Com a utilização da técnica de hole-plate, a bactéria mais sensível observada foi Staphylococcus aureus, com valor de CMI igual ao reportado anteriormente. Foi observado que os extratos alcoólicos obtiveram maior eficiência em relação às infusões, que a técnica de hole-plate revelou ser mais eficiente que a técnica de disco difusão e que os cocos Gram positivos foram mais susceptíveis quando comparadas aos bacilos Gram negativos e fungos.

ABSTRACT Commonly, the urinary treat is sterile, but it can also be contaminated by etiological agents from the intestinal treat, of which the Escherichiacoli is the most common one. These microorganisms are increasingly becoming more resistant to multiple antibiotics. It became necessary to find new substances with antimicrobial properties, so the purpose of this paper is to evaluate different Phyllanthus sp extracts- in face of microorganisms which cause the urinary treat infections- and compare it to the hole-plate and disk diffusion techniques. The 72 hours extraction showed better antimicrobial activity in both methods. Using disk diffusion, the most sensitive bacterium was the Staphylococcus saprophyticcus, with the MIC of 15,84 mg/mL. While using the technique of hole-plate, the most sensitive bacterium was the Staphylococcus aureus, with the same MIC of the previous cited bacterium. This study showed that the alcoholic extracts were more efficient than the infusions. It can also be observed that the hole-plate technique seems to be more efficient than the disk diffusion one, and the Gram positive cocci bacteria were more sensitive than the Gram negative bacilli and fungi.

Efficacy of early treatment on 52 patients with preneoplastic hepatitis B virus-associated hepatocellular carcinoma by compound Phyllanthus Urinaria L / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the change in the number of antibodies of preneoplastic hepatocellular carcinoma (HCC) using early treatment by Compound Phyllanthus Urinaria L. (CPUL) on patients with preneoplastic hepatitis B virus (HBV)-associated HCC.</p><p><b>METHODS</b>A total of 102 cirrhosis patients with regenerative or dysplastic nodules whose sera were tested positive for at least one of these six proteins (five up-regulated genes URG4, URG7, URG11, URG12 and URG19, and one down-regulated gene DRG2) were assigned randomly to two groups using continual random codes by SPSS software. Fifty-two patients were in the treatment group and 50 patients were in the control group. CPUL was used in the treatment group for 3 years, while the control group did not receive any treatment. The changes in HBV-DNA level, number of antibodies, and hepatocarcinogenesis occurred were observed. Patients who did not develop HCC were followed up for another 2 years.</p><p><b>RESULTS</b>HBV-DNA levels decreased ⩾2log in 22.2% (10/45) of patients in the treatment group in contrast to only 5.0% (2/40) of patients in the control group (P=0.0228). The number of antibodies that were tested positive in the treatment group (1.08±1.01) was significantly lower compared with the control group (2.11±1.12) after 24 months of drug treatment (P<0.01). Both the positive rates of anti-URG11 (33/52) and anti-URG19 (31/52) were over 60% at baseline in the two groups, and were decreased to 48.1% (25/52) and 46.2% (24/52) respectively at 36 months of drug treatment, while the rates increased to 68.0% (34/50) and 66.0% (33/50) respectively (P=0.0417, P=0.0436) in the control group. The positive rate of anti-DRG2 was increased to 55.8% (29/52) at 36 months of drug treatment, while in the control group was decreased to 36.0% (18/50, P=0.0452). Among the 102 patients who developed HCC, 2 were in the treatment group and 9 were in the control group, meaning that a significant difference between the two groups (P=0.0212). In 11 patients who developed HCC, anti-URG11 and anti-URG19 were always positive, while anti-DRG2 was negative. Patients newly developing HCC were 6 (20.0%) in the control group, and only one (2.5%) in the treatment group (P=0.0441) during 2-year follow-up after the end of the treatment.</p><p><b>CONCLUSIONS</b>Anti-URG11, anti-URG19 and anti-DRG2 could be used as early markers in the prediction of the therapeutic efficacy of CPUL in treating preneoplastic HCC. CPUL is useful in preventing or delaying the development of HBV-associated cirrhosis to HCC.</p>

Antioxidant effects of Phyllanthus niruri tea on healthy subjects

OBJECTIVE@#To investigate the potential antioxidant effects of Phyllanthus niruri (P. niruri, Euphorbiaceae) tea on healthy subjects.@*METHODS@#Five non-smoking, male healthy volunteers, 20 to 31 years old, were enrolled. Each subject was treated twice, following a randomized crossover fashion regarding the ingestion of P. niruri infusion (5 g/750 mL) (tea group) or 750 mL of water (control group). Fasting venous blood samples were collected prior to and at 1, 2 and 4 h after infusion drinking. Samples were tested for plasmatic gallic acid and ascorbic acid levels, erythrocytic catalase and superoxide dismutase activities, and intracellular DCFH fluorescence in granulocytes, monocytes and lymphocytes.@*RESULTS@#Catalase and superoxide dismutase activities were not altered by tea ingestion. Plasma levels of gallic acid were significantly increased at 1, 2 and 4 h after P. niruri ingestion and plasma ascorbic acid at 1 h after P. niruri ingestion.@*CONCLUSIONS@#Ingestion of P. niruri tea is associated with a slight increase in antioxidant markers in human blood (ascorbic acid and gallic acid), which may contribute to its pharmacological effects.

Atividade antibacteriana in vitro de quatro espécies vegetais em diferentes graduações alcoólicas / In vitro antibacterial activity of four plant species at different alcoholic contents

Neste trabalho foi realizada a caracterização fitoquímica e avaliada a atividade antibacteriana in vitro dos extratos de Ageratum conyzoides L. (mentrasto), Gossypium hirsutum (algodão), Phyllanthus tenellus (quebra pedra), e Polygonum hydropiperoides (erva de bicho) frente à Staphylococcus aureus e Escherichia coli. Para a avaliação da atividade antibacteriana foi utilizado o método de difusão em ágar. Os testes foram realizados com o extrato nas graduações alcoólicas de 0 a 100% (v/v), na proporção de 20% (m/v - massa/extrator). Os testes fitoquímicos constataram a presença de açucares redutores, compostos fenólicos, flavonoides, taninos, triterpenos, e esteróides nas quatro espécies. O crescimento das culturas de S. aureus foi inibido por todos os extratos, com exceção do extrato de Mentrasto. A maior atividade de inibição foi observada pelo extrato de quebra pedra. Entretanto, nenhum dos extratos foi capaz de inibir o crescimento das cepas de E. coli. Os resultados são promissores, visto que três das quatro plantas selecionadas demonstraram possuir substâncias antibacterianas, o que motiva estudos subsequentes para o isolamento e identificação dos princípios ativos responsáveis por essa atividade, com potencial de uso na indústria farmacêutica.

In this study, phytochemical characterization was conducted and the in vitro antibacterial activity of extracts of Ageratum conyzoides L. (whiteweed), Gossypium hirsutum (cotton), Phyllanthus tenellus (shatterstone) and Polygonum hydropiperoides (swamp smartweed) was evaluated against Staphylococcus aureus and Escherichia coli. To assess the antibacterial activity, the agar diffusion method was used. Tests were performed with the extract at alcoholic contents from 0 to 100% (v/v), at 20% proportion (m/v - mass/extractor). Phytochemical tests indicated the presence of reducing sugars, phenolic compounds, flavonoids, tannins, triterpenes and steroids in all four species. The growth of S. aureus cultures was inhibited by all extracts, except for whiteweed extract. The highest inhibitory activity was observed for shatterstone. However, none of the extracts was capable of inhibiting the growth of E. coli strains. Results are promising since three of the four selected plants showed to have antibacterial substances, which stimulates further studies for the isolation and the identification of active principles responsible for this activity, with potential to be used in the pharmaceutical industry.

Isolation, fractionation and evaluation of the antiplasmodial properties of Phyllanthus niruri resident in its chloroform fraction

OBJECTIVE@#To investigate the antiplasmodial activity of Phyllanthus niruri (P. niruri) methanol extract (ME) and its fractions in mice.@*METHODS@#P. niruri methanol extract and its chloroform, ethanol and aqueous portions were tested against chloroquine-sensitive Plasmodium berghei berghei in early, established and repository models of infection using Knight and Peter's 4-day suppressive model, Ryley and Peters curative model and Peters prophylactic model respectively.@*RESULTS@#Chemosuppression of parasitaemia (37.65%-50.53 %) was elicited by 100-400 mg/kg (b.w.) of ME. At doses of 100 mg/kg b.w., the chloroform fraction (F1) significantly (P<0.01) suppressed parasitaemia by 85.29%, while ethanol and aqueous fractions (F2 and F3, respectively) elicited 67.06% and 51.18% chemosuppression. The most active fraction, F1 was selected for further antiplasmodial screening. In established infection, ME reduced parasitaemia (15.81%-62.96%) while F1 significantly (P<0.01) reduced parasitaemia (44.36%-90.48%), with effects comparable to that of chloroquine (96.48%). The prophylactic antiplasmodial activity of ME (92.50% suppression) was also significant (P<0.01) and was more effective than pyrimethamine (85.00%). Additionally, cell membrane integrity of non-parasitized erythrocytes incubated with 125-500 mg/mL F1 was maintained.@*CONCLUSIONS@#These findings indicate the antiplasmodial efficacy of P. niruri methanol extract, and the localization of this effect in its chloroform fraction.

Phytochemical analysis and anticancer capacity of Shemamruthaa, a herbal formulation against DMBA- induced mammary carcinoma in rats

OBJECTIVE@#To investigate the bioactive constituents of Shemamruthaa (SM), a herbal combination and its therapeutic effects on the mitochondrial functions with reference to lipid peroxidation (LPO), antioxidant status, citric acid cycle enzymes and electron transport chain enzymes in mammary tissues of 7,12-dimethylbenz(a)-anthracene (DMBA) induced mammary carcinoma in rat model.@*METHODS@#Adult female Sprague-Dawley rats were used for the study and were divided into four groups. Group I served as control and Group II rats were induced mammary carcinoma by administration of DMBA (25 mg/kg b.w.) orally. The normal and cancer-induced rats (Group III) were treated with SM (400 mg/kg b.w./day) orally by gastric incubation for 14 days. Group IV rats served as SM-treated control animals.@*RESULTS@#Cancer-induced rats showed a considerably increased level of LPO with concomitant decreased levels of antioxidants, citric acid cycle enzymes, electron transport chain enzymes and cytochrome contents in the mammary tissue. Treatment with SM brought back the aforementioned biochemical parameters to near normal.@*CONCLUSIONS@#From the results, it can be inferred that Shemamruthaa possesses significant anticancer effect through its role in attenuation of LPO, prevention of membrane damage and restoring membrane integrity.

Acute toxicity study of stone-breaker (Phyllanthus tenellus Roxb)

Phyllanthus tenellus Roxb. é nativa do Brasil, mais frequentemente em ambientes úmidos, e usada para o tratamento de litíase urinária, doença inflamatória intestinal, diabetes e hepatite B. Neste trabalho objetiva-se determinar a toxicidade aguda e DL50 do extrato aquoso de P. tenellus em animais de laboratório e avaliar o seu comportamento. A DL50 por via intraperitoneal foi calculada pelo método de Karber e Behrens (1964), em que o extrato alcoólico a 96% foi concentrado em evaporador rotativo. Utilizou-se camundongos albinos (Mus musculus) machos, divididos em 3 lotes de seis animais. Eles foram observados por 24 horas a partir da administração do extrato diluído em solução fisiológica a 0,9% nas dosagens de 500; 1000; 1500; 2000; 2500 mg/kg. Estudos de curto prazo têm demonstrado que esta planta não é considerada tóxica, porém, constatamos que esta espécie provoca agitação nos animais por movimentos estereotipados, espasmos, e um aumento da frequência respiratória, bem como ações de depressão, tais como: sonolência, prostração, dispneia e diminuição da frequência respiratória.

Phyllanthus tenellus Roxb. is a herbaceous plant native to Brazil and appears frequently in humid environments. This plant is used to treat urolithiasis, inflammatory bowel disease, diabetes and hepatitis B. The acute toxicity and LD50 of an aqueous extract of P. tenellus were determined in laboratory mice and their behavior was analyzed. The intraperitoneal LD50 was calculated by the Karber & Behrens (1964) method, for which a 96% alcoholic extract was concentrated in a rotary evaporator. Male albino mice (Mus musculus) were divided into three batches of six animals and observed for 24 hours after administration of the extract, diluted in 0.9% saline, at doses of 500, 1000, 1500, 2000 and 2500 mg / kg. Short-term studies have demonstrated this plant to be non-toxic; however, we found that this species induced agitation in animals, with stereotyped movements, spasms and increased respiratory frequency, as well as signs of depression, such as sleepiness, prostration, dyspnea and a reduction in respiratory frequency.

Efecto de Phyllanthus orbicularis sobre la viabilidad celular y el antígeno de superficie de la hepatitis B en células PLC/PRF/5 / Effect of Phyllanthus orbicularis on the cell viability and the hepatitis B surface antigen in PLC/PRF/5 cells

La actividad antiviral de Phyllanthus orbicularis Kunth, planta endémica de Cuba, frente al virus de la hepatitis B, fue estudiada in vitro. Se determinó la capacidad del extracto acuoso y 2 fracciones químicas ricas en taninos y flavonoides (acético-acuosa y butanólica) para inhibir la producción o bloquear la detección del antígeno de superficie de la hepatitis B (AgsHB) en las células PLC/PRF/5, una línea celular de hepatoma humano que expresa este marcador constitutivamente. El extracto acuoso fue efectivo al inhibir el AgsHB producido a las 24 y 48 h con índices selectivos superiores a 10, con una acción inhibitoria dependiente de la concentración. La evaluación de la actividad de las fracciones obtenidas a partir de esta planta mostró que ambas eran efectivas con índices selectivos superiores a 10. Estos resultados sugieren que existe una actividad inhibitoria del antígeno estudiado y que esta planta posee algunos compuestos fenólicos que podrían actuar sobre el AgsHB y reducir su producción en las células, lo que indica una posible actividad in vivo. Este trabajo corrobora el potencial de acción antiviral del extracto acuoso de P orbicularis, además de avalar el empleo de la línea PLC/PRF/5 para futuras evaluaciones o ensayos encaminados a valorar la actividad antihepatitis B de diferentes productos. De igual forma abre el camino para la determinación de aquel o aquellos componentes químicos responsables de la actividad de la planta frente al virus de la hepatitis B

The antiviral activity of Phyllanthus orbicularis, an endemic Cuban plant, against the hepatitis B virus was evaluated in vitro. The ability of the aqueous extract and 2 tannin- and flavonoid-rich chemical fractions (aqueous-acetic and buthanolic) to inhibit the hepatitis B surface antigen (HBsAg) production was determined in PLF/PRC/5 cellas, a human hepatoma cell line that constitutively expresses this antigen. The aqueous extract was effective in inhibiting the HBsAg at 24 and 48 hours of treatment, being the selective index (SI) above 10, with concentration-dependent inhibitory activity. The evaluation of the inhibitory action of the two fractions from the plant proved that both were effective and their selective indexes were also higher than 10. These results indicated the inhibitory activity against HBsAg of the plant and the presence of phenolic compounds that could reduce the antigen production. This may contribute to an in vivo antiviral action. This paper also corroborated the antiviral potential of the aqueous extract from Phyllanthus orbicularis Kunth, in addition to supporting the use of PLC/PRF/5 for future evaluations or trials directed towards assessing the anti-hepatitis B activity of various products. Likewise, it paved the way for the detection of the chemical component(s) responsible for the plant's action against hepatitis B virus

Caracterización química por cromatografía de gases/espectrometría de masas de dos extractos obtenidos de Phyllanthus orbicularis HBK / Chemical characterization using gas chromatography/mass spectrometry of two extracts from Phyllanthus orbicularis HBK

El objetivo del presente trabajo consistió en la caracterización química por cromatografía de gases/espectrometría de masas de 2 extractos obtenidos de Phyllanthus orbicularis HBK, para lo cual se llevó a cabo un método de extracción por maceración con n-hexano y acetato de etilo, a fin de obtener los extractos correspondientes. El estudio del extracto hexánico permitió identificar 17 componentes y resultaron predominantes estructuras del tipo hidrocarbonada, entre ellas, el ciclooctacosano como componente mayoritario. En el extracto de acetato de etilo se identificaron 19 compuestos y constituyeron los terpenoides las estructuras mayoritarias, aunque el más abundante resultó ser el esterol ã-sitosterol. Los compuestos identificados se informan por primera vez para la especie

The objective of this paper was the chemical characterization of two extracts from Phyllanthus orbicularis HBK through gas chromatography/mass spectrometry. To this end, maceration with N-hexane and ethyl acetate was used to obtain the respective extracts. The study of the hexane extract identified 17 components in which hydrocarbonate structures prevailed, mainly cyclooctacosane. In the ethyl acetate extract, 19 compounds were detected, being the terpenoids the predominant, although the most abundant was sterol ã-sitosterol. For the first time, the identified compounds are reported for this species

Densidade de plantio e idade de colheita de quebra-pedra [Phyllanthus amarus (Schumach. & Thonning) genótipo Unicamp-CPQBA 14] na produtividade de filantina / Density of planting and age of Phillanthus amarus genotype UNICAMP/CPQBA-14 on the productivity of phyllanthin

Avaliou-se o efeito da densidade de plantio e da idade de Phyllanthus amarus CPQBA-14, sobre o teor e produtividade de filantina. O delineamento experimental foi em blocos ao acaso, em esquema fatorial 4 x 6, com cinco repetições, envolvendo 4 tratamentos de densidade de plantio (400.000 pl ha -1; 200.000 pl ha -1; 100.000 pl ha -1 e 66.667,68 pl ha -1) com 6 colheitas (30, 45, 60, 75, 90 e 105 dias após o transplante (DAT)). Os dados foram submetidos a Análise de Variâncias e Regressão (P > 0.001). Para o teor de Filantina, foram observados efeitos significativos e independentes dos tratamentos de idade da planta na colheita e densidade de plantio. O teor de filantina aumentou linearmente com a idade da planta, atingindo o máximo aos 105 DAT, com teor de 11,52 g Kg-1. O adensamento populacional estimado que proporcionou máximo teor de filantina (8,66 g kg-1) foi de 299.860 pl ha-1. A produtividade de filantina apresentou interação significativa entre densidade de plantio e idade da planta na colheita. Observou-se que ao longo do crescimento da planta, os tratamentos com 200 e 400.000 pl ha -1 apresentaram as maiores produtividades de filantina (39,3 e 37,8 kg ha-1) aos 97,12 e 95,17 DAT, respectivamente. Conclui-se que o espaçamento apropriado para o melhor arranjo espacial de cultivo é de 299.860 pl ha-1 para as produtividades de 3.974,19 Kg (massa de matéria seca foliar) x 0,00866 kg (teor de filantina) = 34,416 kg ha-1 de filantina, com colheita programada aos 97 DAT.

The effect of planting density and age of Phyllanthus amarus- genotype CPQBA-14, on the content and productivity of phyllanthin were evaluated. The experimental design was randomized blocks in factorial scheme 4 x 6, with five replicates, consisting of 4 treatments of planting density (400.000 pl ha-1; 200.000 pl ha-1; 100.000 pl ha-1 and 66667.68 pl ha-1) with six ages of plants (30, 45, 60, 75, 90 and 105 days after transplanting (DAT)). Data were subjected to analysis of variance and regression (P>0.005). For phyllanthin content independent and significant effects of harvest and planting density were observed. The phyllanthin contents increased with plant age, peaking at 105 DAT, with a content of 11.52 g kg-1. The estimated population density that provided the high phyllanthin contents (8.66 g kg-1) was at 299.860 pl ha-1. The productivity of phyllanthin showed significant interaction between planting density and plants ages. It was observed during the plant growth, the treatments with 200 and 400.000 pl ha -1 had the highest yield of phyllanthin (39.3 and 37.8 kg ha-1) with 95.17 and 97.12 DAT, respectively. We conclude that the proper spacing for best spatial arrangement of the cultivation is 299.860 pl ha-1 for the yield of 3974.19 kg (dry weight leaf) x 0.00866 kg (phyllanthin contents) = 34.416 kg ha-1 of phyllanthin with harvest scheduled at 97 DAT.

The antiplasmodial effect of the extracts and formulated capsules of Phyllanthus amarus on Plasmodium yoelii infection in mice

OBJECTIVE@#To investigate the antiplasmodial activity of the extracts of Phyllanthus amarus (P. amarus) on Plasmodium yoelii (P. yoelii) (a resistant malaria parasite strain used in animal studies) infection in mice.@*METHODS@#The aqueous and ethanol extracts of the whole plant of Phyllanthus amarus was administered to Swiss albino mice at doses of 200 mg/kg/day, 400 mg/kg/day, 800 mg/kg/day and 1600 mg/kg/day and the prophylactic and chemotherapeutic effect of the extracts against P. yoelii infection in mice was investigated and compared with those of standard antimalaria drugs used in the treatment of malaria parasite infection. Acute toxicity test was carried out in mice to determine the safety of the plant extract when administered orally.@*RESULTS@#The results showed that the extracts demonstrated a dose-dependent prophylactic and chemotherapeutic activity with the aqueous extracts showing slightly higher effect than the ethanol extract. The antiplasmodial effects of the extracts were comparable to the standard prophylactic and chemotherapeutic drugs used in chloroquine resistant Plasmodium infection although the activity depended on the dose of the extract administered. The extracts showed prophylactic effect by significantly delaying the onset of infection with the suppression of 79% at a dose of 1600 mg/kg/day.@*CONCLUSIONS@#The results obtained indicate that the extracts of the whole plant of P. amarus possess repository and chemotherapeutic effects against resistant strains of P. yoelii in Swiss albino mice. The findings justify the use of the extract of P. amarus in traditional medicine practice, for the treatment of malaria infections.

Protective effects of Phyllanthus acidus (L.) Skeels leaf extracts on acetaminophen and thioacetamide induced hepatic injuries in Wistar rats

OBJECTIVE@#To investigate and compare the hepatoprotective effects of crude ethanolic and aqueous extracts of Phyllanthus acidus (L.) Skeels (P. acidus) leaves on acetaminophen (APAP) and thioacetamide (TAA) induced liver toxicity in wistar rats. Silymarin was the reference hepatoprotective agent.@*METHODS@#In two different sets of experiments, the P. acidus extracts (200 and 400 mg/kg, body weight) and silymarin (100 mg/kg, body weight) were given orally for 7 days and a single dose of APAP (2 g/kg, per oral) or TAA (100 mg/kg, subcutaneous) were given to rats. The level of serum aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total bilirubin and total protein were monitored to assess hepatotoxicity and hepatoprotection.@*RESULTS@#APAP or TAA administration caused severe hepatic damage in rats as evident from significant rise in serum AST, ALT, ALP, total bilirubin and concurrent depletion in total serum protein. The P. acidus extracts and silymarin prevented the toxic effects of APAP or TAA on the above serum parameters indicating the hepatoprotective action. The aqueous extract was found to be more potent than the corresponding ethanolic extract against both toxicants. The phenolic and flavonoid content (175.02±4.35 and 74.68±1.28, respectively) and 2,2-diphenyl-1-picrylhydrazil (DPPH) [IC(50) = (33.2±0.31)μg/mL] scavenging potential was found maximum with aqueous extract as compared to ethanolic extract.@*CONCLUSIONS@#The results of present study suggests that the aqueous extract of P. acidus leaves has significant hepatoprotective activity on APAP and TAA induced hepatotoxicity, which might be associate with its high phenolic and flavonoid content and antioxidant properties.