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Analysis of the mechanisms underlying the contractile response induced by the hydroalcoholic extract of Phyllanthus urinaria in the guinea-pig urinary bladder in-vitro.

The hydroalcoholic extract of Phyllanthus urinaria (Euphorbiaceae), substance P and substance P methyl ester all caused graded contractions in the guinea-pig urinary bladder. Responses to hydroalcoholic extract and substance P were markedly inhibited in calcium-free Krebs solution, this effect being reversed by reintroduction of calcium in the medium. The contraction in response to hydroalcoholic extract was unaffected by atropine, propranolol, prazosin, yohimbine, tetrodotoxin, w-conotoxin, nicardipine, HOE 140, guanethidine, staurosporine, phorbol ester or indomethacin, excluding the involvement of nervous mediated responses, or action via cholinergic, adrenergic, kinins, cyclo-oxygenase metabolites, protein kinase C or activation of L or N-type calcium channels. The selective NK1 tachykinin antagonist (FK 888), but not NK2 (SR 48968) antagonized substance P-induced contraction, but both drugs failed to effect Phyllanthus urinaria-induced contraction. Prolonged desensitization of guinea pig urinary bladder with capsaicin (10 microM) or preincubation of guinea-pig urinary bladder with capsazepine did not affect contraction caused by hydroalcoholic extract. Ruthenium red almost completely abolished capsaicin-induced contraction, but had no effect on hydroalcoholic extract-mediated contraction. Substance P and the hydroalcoholic extract caused marked potentiation of the twitch response in the preparations field stimulated. The facilitatory effect of substance P, but not that of hydroalcoholic extract, was prevented by the NK1 (FK 888), but not by NK2 (SR 48968) antagonist. We concluded that contraction induced by hydroalcoholic extract of Phyllanthus urinaria in the guinea pig urinary bladder involves direct action on smooth muscle and relies on the mobilization of extracellular calcium influx unrelated to activation of L- and N-type calcium channels or activation of protein kinase C mechanisms. In addition contraction caused by the hydroalcoholic extract of Phyllanthus urinaria in guinea-pig urinary bladder does not involve the activation of tachykinin or vanilloid receptors.

Adrenergic alpha-Antagonists↗

The in vitro activity of geraniin and 1,3,4,6-tetra-O-galloyl-beta-D-glucose isolated from Phyllanthus urinaria against herpes simplex virus type 1 and type 2 infection.

Phyllanthus urinaria Linnea (Euphorbiaceae) is a widely used traditional medicinal plant by oriental countries and has been reported to possess various biological activities. Previously, the acetone extract from Phyllanthus urinaria was found to inhibit herpes simplex virus (HSV) infection. In this study, geraniin and 1,3,4,6-tetra-O-galloyl-beta-D-glucose (1346TOGDG), both of which were isolated from the acetone extract of Phyllanthus urinaria, were examined for their activity against HSV-1 and HSV-2 in vitro. Results showed that geraniin actively suppressed HSV-2 infection, whereas 1346TOGDG effectively inhibited HSV-1 infection. The 50% inhibitory concentration (IC(50)) was 18.4+/-2.0 microM for geraniin against HSV-2 infection, and 19.2+/-4.0 microM for 1346TOGDG against HSV-1. No toxic effect towards the host cell was observed at the antiviral concentrations. In conclusion, geraniin and 1346TOGDG were found to inhibit HSV-1 and HSV-2 multiplication at different magnitudes of potency.

Animals↗

[Microscopy identification of Phyllanthus simplex].

OBJECTIVE: To identify Phyllanthus simplex. METHODS: Morphological and histological identification. RESULTS: There are cluster crystals in cortex and mesophyll of the root and the stem from Phyllanthus simplex. Paracytic type stomata are visible. CONCLUSION: Above-mentioned characters can be used for the identification of Phyllanthus simplex.

Pharmacognosy↗

Modification of metal-induced micronuclei formation in mouse bone marrow erythrocytes by Phyllanthus fruit extract and ascorbic acid.

The comparative efficacy of Phyllanthus emblica fruit extract, and its main constituent, ascorbic acid, in modifying the cytotoxic action of lead and aluminium was evaluated in vivo in bone marrow cells of Mus musculus. Oral administration of Phyllanthus emblica fruit extract for 7 days before exposure to the metals through intraperitoneal injections reduced the frequency of micronuclei induced by all doses of both metals. Priming with comparable doses of synthetic ascorbic acid reduced micronuclei formation induced by both doses of aluminium and only the lower dose of lead. With the higher dose of lead (20 mg/kg body wt.) priming with ascorbic acid increased the frequency of micronuclei when compared with mice administered lead alone. The greater efficacy of Phyllanthus fruit extract in alleviating metal-induced clastogenicity may be due to the combined action of all ingredients in the crude extract, rather than to ascorbic acid alone.

Aluminum↗

Usage and bioassays in Phyllanthus (Euphorbiaceae). IV. Clustering of antiviral uses and other effects.

A number of species of the genus Phyllanthus (Euphorbiaceae) have been tested for their efficacy as antivirals, partly on the basis of references to traditional usage for the treatment of diseases possibly having a viral origin. There are also many references to indigenous uses and to laboratory assays for other biological activities in this large genus (550+ species). These citations have been arranged by subgenus, section, subsection and species and have been published in three previous papers. This paper summarizes selected clustering of usage and effect by subgeneric taxa. Consideration of the data from ethnobotany, in vitro assays and clinical trials supported the presence of some type of biological activity(s) particularly within the subgenus Phyllanthus. Although the herbaceous species of subgenus Phyllanthus have been extensively used to treat jaundice, and have generally inhibited hepadnavirus DNAp, effects on chronic infection with hepatitis B virus (HBV) or related viruses have generally been negative. Alternative explanations for the wide usage have been little explored. Other medical categories suggested possible leads for research, or possibly, herbal or galenic remedies with bona fide effects. In most cases, the data remain suggestive but not conclusive.

Animals↗

[Mechanism of protective action of Phyllanthus urinaria L. against injuries of liver cells].

It has been found out that the carbon tetrachloride (CCl4)-induced increase of serum glutamic-pyruvic transaminase (ALT) and elevation of MDA in liver of mice are significantly lowered by Phyllanthus urinaria in vivo, and the coincubation of isolated rat hepatocytes with Phyllanthus urinaria in vitro significantly inhibits CCl4-induced decrease of mobility of membrane of liver cells and increase of intracellular free Ca2+ ([Ca2+]i) concentrations of liver cells. These results suggest that the anti-lipid peroxidation effect and protective action of membrane of Phyllanthus urinaria may be related to its protective action against CCl4-induced liver injuries.

Alanine Transaminase↗

Analysis and identification of lignans in Phyllanthus urinaria by HPLC-SPE-NMR.

HPLC-SPE-NMR has been applied to the analysis of a number of structurally similar lignans present in Phyllanthus urinaria. A reversed-phase HPLC system, using a C8 column eluted with tetrahydrofuran-water-methanol and detected at 225 nm was developed that gave good resolution for seven lignans from Phyllanthus urinaria. Coupling of this system with SPE-NMR provided clean 1H-NMR spectra for nine lignans present in 4 mg of a lignan-rich fraction, equivalent to ca. 1.0 g of dry plant material. The result demonstrates that HPLC-SPE-NMR is a very efficient and powerful tool for the unambiguous identification of natural products that have been previously characterised.

Chromatography, High Pressure Liquid↗

Quantitative determination of phyllanthin and hypophyllanthin in Phyllanthus species by high-performance thin layer chromatography.

A simple, precise and rapid high-performance thin-layer chromatographic method has been developed for the estimation of phyllanthin (1) and hypophyllanthin (2), the important lignans of Phyllanthus species, especially Phyllanthus amarus. Separation of 1 and 2 was carried out on silica gel 60 F254 layers eluted with hexane:acetone:ethyl acetate (74:12:8), and the analytes were visualised through colour development with vanillin in concentrated sulphuric acid and ethanol. Scanning and quantification of spots was performed at 580 nm. Recoveries of 1 and 2 were 98.7 and 97.3%, respectively. The method was validated and the peak purities and limits of detection and quantification were determined.

Chromatography, Thin Layer↗

Some clinico-pathological changes associated with the aqueous extract of the leaves of Phyllanthus amarus in rats.

The pathological changes of the aqueous crude extract of the leaves of Phyllanthus amarus were studied in 32 male rats over a period of 30 days. The animals were divided into four groups of eight animals per group. The aqueous crude extract was prepared and administered orally using a cannula to rats in three groups receiving doses of 400 mg/kg, 800 mg/kg and 1,000 mg/kg but the fourth group served as a control and received distilled water only. Blood samples were collected for haematological and serum biochemical analysis. Organs such as the liver, kidney, testes and pancreas were also assessed for histopathological changes. The study showed that the extract caused a decrease in the red blood cell (RBC) count, packed cell volume (PCV), haemoglobin concentration (Hb), but an increase in the white blood cell (WBC) count. The extract also resulted in an increase in the levels of aspartate amino transferase (AST), total and conjugated bilirubin, total protein and albumin. The study, however, caused a decrease in the level of alanine amino transferase (ALT). Histopathologically, there were cases of protein casts in the kidney tubules with tubular nephrosis, foci of lymphocytic infiltration at the portal areas of the liver as well as marked testicular degeneration with severe disorganization of seminiferous tubules, which were devoid of spermatic cells. A reduction in the weight of the experimental animals was also noted in this study. It thus shows that Phyllanthus amarus has potential toxic properties.

Animals↗

Anti-mutagenic activity of Phyllanthus amarus Schum & Thonn in vitro as well as in vivo.

Methanolic extract of Phyllanthus amarus was tested for its anti-mutagenic activity in Salmonella typhimurium strains TA1535, TA100, and TA102 (Ames test). P. amarus extract was able to inhibit the activation and mutagenicity of 2-acetaminofluorene (2-AAF) and aflatoxinB(1) at concentrations of 0.25-2 mg/plate. It was also found to inhibit mutagenicity induced by direct acting mutagens sodium azide (NaN(3)), N-methyl-N-nitro-N-nitrosoguanidine (MNNG), and 4-nitro-0-phenylenediamine (NPD), at concentrations of 1 mg to 0.25 mg/plate. Urinary mutagenicity produced in rats by benzo[a] pyrene was found to be significantly inhibited by the oral administration of Phyllanthus extract. These results indicate significant anti-mutagenicity of the extract in vitro as well as in vivo.

2-Acetylaminofluorene↗

Modulation of rat and human cytochromes P450 involved in PhIP and 4-ABP activation by an aqueous extract of Phyllanthus orbicularis.

Phyllanthus orbicularis HBK (Euphorbiaceae) is a medicinal plant, endemic to Cuba, whose aqueous extract has proven antimutagenic effects against hydrogen peroxide and some promutagenic aromatic amines (AAs), in addition to its antiviral properties. In this paper, antimutagenesis of this extract against two carcinogenic AAs, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 4-aminobiphenyl (4-ABP) has been studied. Liver microsomal fractions from both induced rats and humans were used to metabolise both procarcinogenic compounds in the Salmonella assay. The plant extract was effective in reducing the mutagenesis of these AAs, activated by both kinds of fractions. The optimal antimutagenic effect was obtained when both AAs were metabolised by human enzymes, with an almost total reduction of 4-ABP mutagenesis and a decrease of about 75% of PhIP mutagenicity. Mutagenicity of both AAs, activated by induced rat fraction, was only decreased by about 50%. Inhibition by plant extract of alkoxyresorufin O-dealkylation activities, dependent on CYP1A, of both fractions was determined. In accordance with the results obtained, the inhibition or modulation of CYP1A subfamily activities, and possibly of CYP1A2, is thought to be the main mechanism of antimutagenesis of the aqueous extract of Phyllanthus orbicularis against 4-ABP and PhIP.

Aminobiphenyl Compounds↗

Protective effect of Phyllanthus fraternus against allyl alcohol-induced oxidative stress in liver mitochondria.

The effect of administration of allyl alcohol on the oxidative stress and the protective effect due to administration of an aqueous extract of Phyllanthus fraternus against allyl alcohol-induced damage in liver mitochondria were studied. When rats were treated with allyl alcohol, the rate of mitochondrial respiration was decreased significantly with both NAD(+)- and FAD-linked substrates. The respiratory control ratio, an index of membrane integrity and the P/O ratio, a measure of phosphorylation efficiency also decreased significantly. There was a significant increase in the lipid peroxide level and the protein carbonyl content. A significant decrease was observed in the total sulphydryl groups and a significant increase in the generation of superoxide radicals. Administration of rats with an aqueous extract of Phyllanthus fraternus (100 mg/kg) prior to allyl alcohol administration showed protection of 72, 40 and 80% using glutamate+malate (NADH oxidation) and 77, 54 and 20% using succinate as substrate on state 3, RCR and P/O ratio, respectively. The protection on lipid peroxide level was 88 and 91% in homogenate and mitochondria, respectively. In case of protein carbonyls, total sulphydryl groups and on the generation of superoxide radicals the protection was 99, 59 and 53%, respectively.

Animals↗

alpha-Amylase inhibitory activity of some Malaysian plants used to treat diabetes; with particular reference to Phyllanthus amarus.

Extracts of six selected Malaysian plants with a reputation of usefulness in treating diabetes were examined for alpha-amylase inhibition using an in vitro model. Inhibitory activity studied by two different protocols (with and without pre-incubation) showed that Phyllanthus amarus hexane extract had alpha-amylase inhibitory properties. Hexane and dichloromethane extracts of Anacardium occidentale, Lagerstroemia speciosa, Averrhoa bilimbiPithecellobium jiringa and Parkia speciosa were not active when tested without pre-incubation. Extraction and fractionation of Phyllanthus amarus hexane extract led to the isolation of dotriacontanyl docosanoate, triacontanol and a mixture of oleanolic acid and ursolic acid. Dotriacontanyl docosanoate and the mixture of oleanolic acid and ursolic acid are reported from this plant species for the first time. All compounds were tested in the alpha-amylase inhibition assay and the results revealed that the oleanolic acid and ursolic acid (2:1) mixture was a potent alpha-amylase inhibitor with IC(50)=2.01 microg/ml (4.41 microM) and that it contributes significantly to the alpha-amylase inhibition activity of the extract. Three pure pentacyclic triterpenoids, oleanolic acid, ursolic acid and lupeol were shown to inhibit alpha-amylase.

Drugs, Chinese Herbal↗

Phylogeny of medicinal Phyllanthus species in China based on nuclear ITS and chloroplast atpB-rbcL sequences and multiplex PCR detection assay analysis.

Plants of the genus Phyllanthus are commonly used in India, China and Korea for medicinal purposes. Although they are widely cultivated and marketed, there has been uncertainty about the efficacy of different species. A prerequisite of Good Agricultural Practice (GAP) is the authentication of relevant species, and this is now made unequivocal by applying DNA sequence tools. In this study the phylogenetic relationships among 18 Phyllanthus species found in China were investigated by DNA sequence analyses of the nuclear internal transcribed spacers (ITS1 and ITS2) along with the combined chloroplast ATPB and RBCL sequences. Cladistic analysis indicated that this genus is paraphyletic and strongly supports the notion that two problematic and confusing species, P. niruri and P. amarus, are two individual, albeit closely related, species. We have also developed an ITS rDNA-based multiplex PCR assay to differentiate three medicinally important species, P. amarus, P. niruri and P. urinaria.

DNA, Chloroplast↗

[Studies on chemical constituents of Phyllanthus urinaria L].

OBJECTIVE: To investigate the chemical constituents of Phyllanthus urinaria. METHOD: Various chromatographic techniques were employed for isolation and purification of the constituents. The structures were elucidated by chemical and spectral analyses. RESULT AND CONCLUSION: Fourteen compounds were isolated and seven of them were identified as corilagin(I), rutin(II), brevifolincarboxylic acid(VI), isostrictiniin(IX), geraniin (X), gallic acid(XI) and ellagic acid (XII). Compound VI was found from P. urinaria for the first time and compound IX was found from genus Phyllanthus for the first time.

Carboxylic Acids↗

[Preliminary evaluation of the antiviral activity of the aqueous extract of Phyllanthus orbicularis vs HIV-1 infection].

The antiviral activity of the aqueous extract of Phyllanthus orbicularis, a member of the Euohorbiacea family, against the simple herpes virus typel was evaluated in cellular culture, specifically in fibroblasts of human prepucium, and in an animal model, Balb-c mice. The extracelullar effect of the extract on HIV-1 proved to be effective. A selective index of 44 was obtained, which shows a possible virucidal action on the particle. In the trial in vivo, the topical administration of the aqueous extract (12 mg/kg) reduced significantly the development of lesions in mice subcutaneously infected with HIV-1 (1 x 106 UFP). These results suggest the consideration of Phyllanthus orbicularis as a possible anti-HIV-1 candidate

Animals↗

Effect of Phyllanthus amarus on duck hepatitis B virus replication in vivo.

Nine ducks congenitally infected with the duck hepatitis B virus (DHBV) were treated either orally (four ducks for 10 weeks) or intraperitoneally (five ducks for 12 weeks) with the Indian traditional herbal remedy Phyllanthus amarus. Compared to placebo-treated control ducks, these treatments did not result in a reduction of circulating viral DNA in the serum or in the level of viral DNA replication in the liver. In two of the five intraperitoneal-treated ducks, a reduction in the levels of duck hepatitis B surface antigenaemia (DHBsAg) was observed. The data strongly suggest that Phyllanthus amarus has no significant inhibitory effect on DHBV DNA replication and only a minor effect on DHBsAg production.

Animals↗

The effect of Phyllanthus amarus aqueous extract on blood glucose in non-insulin dependent diabetic patients.

The glycaemic response to 124.5 +/- 9.3 (mean +/- SD) g of pancakes was monitored in 21 non-insulin dependent diabetic (NIDDM) patients while on oral hypoglycaemics, after a 1-week washout period and after a 1-week twice daily treatment with 100 mL of an aqueous extract from 12.5 g of powdered aerial parts of Phyllanthus amarus. After the 1-week washout period, the fasting blood glucose (FBG) and postprandial blood glucose increased significantly compared with treatment on oral hypoglycaemics ( p < 0.05). After a 1-week herbal treatment no hypoglycaemic activity was observed. Both FBG and postprandial blood glucose remained very similar to that recorded after the washout period ( p > 0.05). Both liver and renal functions based on alanine transaminase (ALAT) and serum creatinine, respectively, were not significantly affected by the use of the extract. Although the lymphocyte and monocyte levels were significantly decreased ( p < 0.05) and the granulocyte level was significantly increased after treatment ( p < 0.05) the overall total white blood cell (WBC) count and haemoglobin (Hb) were not significantly affected by the 1 week herbal treatment. We conclude that 1 week treatment with the aqueous extract of Phyllanthus amarus was incapable of lowering both FBG and postprandial blood glucose in untreated NIDDM patients.

Adult↗