Extract Of Phyllanthus Urinaria Research Articles

The Effect of Various Sterilization Methods and Volume Containers on Phytochemical Content of Methanol Extract of Phyllanthus urinaria

Phyllanthus urinaria is an annual perennial herbal species found in tropical Asia, America, China, and the Indian Ocean islands. Phyllanthus urinaria is used in folk medicine as a cure to treat jaundice, diabetes, malaria, and liver diseases. Sterilizing the substrate is a crucial step in the fermentation process. This process ensures that the inoculated microorganism is entirely single. Autoclave sterilization is widely favored within the scientific community. In autoclaving, pressurized steam is employed to deliver heat, effectively reducing the bioactive compounds present in the substrate. Comparative studies on various sterilization methods have been reported. This study aims to investigate the effects of substrate containers in sterilization methods of the herbal plant on phenol and flavonoid compounds by LC-MS (Liquid Chromatography-Mass Spectrometry) analysis. Three sterilization methods (pasteurization, steam, and autoclave sterilization) were each applied to the meniran herbal plant (Phyllanthus urinaria) for 15 minutes. Using the aluminum chloride colorimetric assay, the sterilization results were measured for total phenol content, the Folin-Ciocalteu test, and total flavonoid content. The LC-MS analysis showed that the methanol extract of Phyllanthus urinaria (APU) sterilized by autoclaving resulted in the most significant reduction in active phenolic and flavonoid compounds. Pasteurization, steaming, and autoclaving in a big container resulted in total flavonoid content of 1.80±0.034, 1.70±0.021, and 1.71±0.029 mg QE/g extract. The total phenolic content was 26.49±0.591, 22.77±0.230, and 22.097±0.155 mg GAE extract/g, respectively. Meanwhile, using a small container, each method produced a total flavonoid content of 1.73±0.024, 1.71±0.051, and 1.62±0.015 mg QE/g extract, respectively. The total phenolic content was 20.56±0.093, 19.79±0.295, and 20.09±0.124 mg GAE/g extract. Furthermore, the LC-MS profile revealed that APU experienced a reduction in ρ-hydroxybenzaldehyde and naringenin compounds, leading to a decrease in rutin, methyl brevifolincarboxylate, and ethyl gallate compounds. From the results of LC-MS analysis, this research determined that pasteurization using a big container is the most effective sterilization method for preserving the highest levels of total flavonoid and phenolic content in Phyllanthus urinaria while minimizing adverse effects on phytochemical compounds.

Determination of Phytochemical Composition and the Effects of Ethanol Extract of Phyllanthus urinaria on the Liver and Kidney Function Parameters in Paracetamol-Administered Albino Rat Models

The current study was aimed at determining the phytochemical composition and the effects of Phyllantus urinaria Ethanol extract on the liver and kidneys by using albino rats as the animal models. All the analyses were carried out following standard laboratory methods and procedures. A total of fifteen rats were randomly distributed into three groups of five rats in each group where group one served as the control while the test groups comprise of 1000mg/kg body weight (b.w) and 500mg/kg b.w administered daily for fourteen days and followed by administration of a high dose of paracetamol (200mg/kg b.w) to challenge the system toxicologically. This was deliberately done to note whether the extract having administered for many days will protect the liver and kidneys against the paracetamol by determining the serum biomarkers. The results showed the presence of saponins (8.84 g/100g), tannins (1.32 g/100g), phenols (0.11 g/100g), flavonoids (0.512 g/100g) and alkaloids (0.038 g/100g) while cardiac glycosides, Resins, terpenoids and steroids were not detected. The activity of ALP was significantly (p < 0.05) higher in both the 1000mg/kg b.w and 500mg/kg b.w. There was no significant (p>0.05) decrease in ALT activity of 1000mg/kg b.w and at 500mg/kg b.w. the activity of ALT significant (p>0.05) increased when compared with the control. There was a significant (p<0.05) decrease in AST activity of 1000mg/kg b.w and 500mg/kg b.w when compared with the control. Bilirubin concentration significantly (p< 0.05) decreased at 1000mg/kg b.w and non-significantly (p>0.05) increased at 500mg/kg b.w. ALB significantly (p > 0.05) increased at 1000mg/kg b.w and 500mg/kg b.w. A non-significant decrease in Serum HCO3- in both 1000mg/kg/b.w and 500mg/kg b.w. Urea non-significantly decreased at both 1000mg/kg b.w and 500mg/kg b.w. Creatinine significantly increased at both 1000mg/kg b.w and 500mg/kg b.w concentration when compared to the control. Na+ showed a non-significant increase at 1000mg/kg b.w and 500mg/kg b.w when compared to the control. The extract may not have protected the liver and kidneys against paracetamol despite the presence of some phytochemicals, a histological examination of the liver and kidneys is therefore recommended so as to have a clearer picture of the effects observed.

DEVELOPMENT, CHARACTERIZATION AND ANTIBACTERIAL PROPERTIES OF SILVER NANOPARTICLES LOADED SODIUM ALGINATE/XANTHAN GUM MICROBEADS FOR DRUG DELIVERY APPLICATIONS

Objective: The aim of this study is to create pH-responsive drug carriers, which are useful because they have the potential to improve treatment efficacy by controlling the release rate of ofloxacin from the polymer matrix. Methods: In the first step, silver nanoparticles (Ag NPs) were synthesized from silver nitrate using leaf extract of phyllanthus urinaria L as a reducing agent. In the second step, Ag-NPs-loaded polymeric microbeads were synthesized using sodium alginate (SA) and xanthan gum (XG) for controlled release of ofloxacin (OFLX). The developed microbeads were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (X-RD), transition electron microscopy (TEM), Selected Area Electron Diffraction (SAED), and scanning electron microscopy (SEM). Swelling and in vitro release studies were performed at pH 2.0 and 7.4 at 37 °C. The in vitro antibacterial activity of microbeads were tested against S. mutans, K. pneumoniae, and B. subtilis. The release kinetics and mechanism were analyzed by fitting the release data into different kinetic models and the korsmeyer-peppas equation. Results: FTIR confirms the generation of silver nanoparticle and also the generation of polymeric microbeads. SEM studies reveal the developed microbeads are spherical in shape with rough surfaces. TEM studies reveal the size of 20-40 nm. XRD analysis reveals the molecular dispersion of DOX and the presence of silver nanoparticles in the polymeric matrix. Investigations of in vitro release and swelling studies show that the developed microbeads are relatively suitable for intestinal drug delivery because higher release rate was observed at pH 7.4. The developed microbead follows non-Fickian diffusion drug release mechanism. The created samples exhibited antimicrobial activity against S. mutans, K. pneumoniae, and B. subtilis. Conclusion: The results indicate that microbeads containing OFLX and silver nanoparticles are effective drug-delivery vehicles. A further warrant is required for the use of manufactured microbeads in drug delivery applications.

Expression of IL-6 on Breast Cancer Mice Treated by Combination of Phyllanthus Urinaria and Catharanthus roseus Extract

Breast cancer is one kind of cancer that can cause the death in women. Although, many kind of treatment have done to suppress the growth of breast cancer case, but the number of case still increased. Chemotherphy is one kind of treatment used to suppress breast cancer case. But it is known that can give many poor effect. So it need to get an alternative treatment such as using a herbal plant as medicine as Phyllanthus urinaria and Catharanthus roseus. This research aimed to study the effect of Phyllanthus urinaria and Catharanthus roseus on IL-6 expressed by CD11bin breast cancer model.this research using RAL divided into 6 group as Normal group (N), cancer group (K), Cisplatin treatment (C), Dose 1 (D1) (Phyllanthus urinaria 500 mg/kg weight and Catharanthus roseus 15 mg/kg weight), dose 2 (D2) (Phyllanthus urinaria 1000 mg/kg weight and Catharanthus roseus 75 mg/kg weight) and dose 3 (D3) (Phyllanthus urinaria 2000 mg/kg weight and Catharanthus roseus 375 mg/kg weight). Â the mice had inject by DMBA in 0,015 mg/kg weight in six week. The combination of Phyllanthus urinaria and Catharanthus roseus were administrated orally in two weeks. The expression of IL-6 expressed by CD11b ws analysed by flow cytometry. Data analysed by SPPSS in one way ANOVA and tukey test. Based on result, it showed that dose 3 is the optimal dose to suppress the growth of cancer cells based on IL-6 expression. All of the dose used can suppress the growth of IL-6. Â Â Keywords: Breast cancer, Catharanthus roseus, IL-6, machrophage, Phyllanthus urinaria

Expression of IL-17 on Breast Cancer Mice Treated by Combination of Phyllanthus Urinaria and Catharanthus roseus Extract

This study aimed to evaluate the effect of combination Phyllanthus urinaria and Catharanthus roseus in breast cancer mice based on the expression of IL-17. 7,12-Dimethylbenz(α)anthracene (DMBA) was injected intraperitoneally into normal mice at dose 1.5 mg.kg-1 weight to obtain breast cancer mice. Total of 24 experimental mice divided into normal mice (N), breast cancer mice (K), breast cancer mice with cisplatin (C) treatment (5 mg.kg-1 weight), breast cancer mice with combination extract Dose 1 (P. urinaria 500 mg.kg-1 weigt + C. roseus 15mg.kg-1 weight), breast cancer mice with combination extract Dose 2 (P. urinaria 1000 mg.kg-1 weight + C. roseus 75mg.kg-1 weight), and breast cancer with combination extract Dose 3 (P. urinaria 2000 mg.kg-1 weight + C. roseus 375 mg.kg-1 weight). Cheral was given orally for 14 days. The level of IL-17 was evaluated by flow cytometry analysis. The combination can suppress the expression of IL-17 which down regulation of IL-17 indicate a good prognosis for the breast cancer mice, for 6.17% in breast cancer condition to 0.93% with Dose 3 treatment. The combination can be used as immunomodulatory agent in humoral immunity through the regulation of IL-17. Keywords: Breast cancer, Catharanthus roseus, IL-17, Phyllanthus urinaria

Effect Combination of Catharanthus roseus and Phyllanthus urinaria Extract on Mice BALB/c Breast Cancer Models Based on T-regulator Cells Expression

CD4+CD25+FOXP3+ is a regulatory T cell that plays a role in the body's tolerant system to control activated effector cells. T regulatory plays an important role as a trigger for tumor progressiveness, where an increase in the amount of T regulatory FOXP3 in the tumor state correlates with a poor prognosis. Breast cancer is a type of cancer that tends to increase every year. The purpose of this study was to study the influence of a combination of Phyllanthus urinaria and Catharanthus roseus plant extracts on CD4+CD25+FOXP3+ modulation. Mice was injected subcutaneously with the carcinogen compound DMBA for six weeks at 0.015 mg/kg weight. The study design was conducted using a complete random design (RAL) which was divided into six groups namely healthy (normal) control, cancer control (DMBA injection), Cisplatin and a combination of extracts with three different doses (dose 1 (Phyllanthus urinaria 500 weight and Catharanthus roseus 75mg/kg weight) and dose 3 (Phyllanthus urinary 2000mg/kg weight and Catharanthus roseus 375mg/kg weight). The relative number of T regulatory cells is measured using flow cytometry analysis. The measurements were made using the Cell Quest ProTM program. Statistics analyze using one way ANOVA (p<0.05). Tukey HSD analysis with SPSS 26 software used to analyze real differences between treatments. A relative increase in CD4+CD25+FOXP3+ occurs in cancer control. While the decrease occurred in every treatment except in the dose group 1. The optimal dose to reduce CD4+CD25+FOXP3+ amounts significantly occurred in the dose group 3. This indicates the influence of a combination of Phyllanthus urinaria and Catharanthus roseus on CD4+CD25+FOXP3+ differentiation. Keywords: breast cancer, Catharanthus roseus, CD4+CD25+FOXP3+, Phyllanthus urinaria,

Extraction, Preparation and Evaluation of Phyllanthus urinaria extract as Antifungal Cream

The aim of research work is to formulate an antifungal cream. The cream which is prepared is isolated from natural source. The leaves of Phyllanthus urinaria have active antifungal activity. The genus Phyllanthus is one of the most important groups of plants belonging to the Phyllantaceaceae family. Phyllanthus urinaria is an annual perennial herbal species found in tropical Asia, America, China ad Indian ocean island. P. urinaria is used in folk medicine as a cure to treat jaundice, diabetes, malaria, cancer, bacterial infection, fungal infection. This research gives the therapeutical study for the P. urinaria. The extract is isolated with help of ethanol. The cream is prepared with various pharmaceutical ingredient. Antifungal activity is done with the Aspergillus niger, Candida albicans. This cream is then examined and stability test is performed, physicochemical reactions are done to determine chemical constituents present in the extract.

Determination of Main Compositions in Phyllanthus Urinaria and its Effects on Cyp450 in Rats

<P>Background: Phyllanthus urinaria, a traditional herbal medicine, has aroused widespread concern at home and abroad. However, there are few studies on the effects of Phyllanthus urinaria on CYP450. Therefore, this study aims to explore the main chemical compositions of Phyllanthus urinaria and its effect on the activity of CYP450 enzyme in rats. </P><P> Methods: Acetonitrile and 0.1% Trifluoroacetic Acid (TFA) were used as mobile phase, along with the application of gradient elution to simultaneously determine the main chemical constituents in Phyllanthus urinaria by HPLC (r2>0.999). Sprague-Dawley (SD) rats, randomly divided into control group, low-dose group and high-dose group, were treated with normal saline and different doses of Phyllanthus urinaria extract solution, respectively. Additionally, the rats were given intragastric administration of cocktail probe (specific substrates of CYP450 isoenzyme) at 15th day; the plasma was collected by tail vein at various times. Furthermore, the UPLC-MS/MS method (r2>0.99) was used to detect the probe concentration, along with the evaluation of the activity of CYP450 enzyme according to the pharmacokinetic parameters of the probe. </P><P> Results: Gallic acid, 3, 4-dihydroxybenzoic acid, caffeic acid, corilagin and ellagic acid were found in the Phyllanthus urinaria extract solution by HPLC. Compared with the control group, the metabolism of bupropion, metoprolol, midazolam and tolbutamide slowed down significantly in the Phyllanthus urinaria group, with no significant metabolic changes in phenacetin. </P><P> Conclusion: Phyllanthus urinaria could induce activity of CYP2D6, CYP2B1, CYP3A4 and CYP2C9, without exerting a significant effect on CYP1A2.</P>

Sử dụng thức ăn bổ sung diệp hạ châu (Phyllanthus urinaria) phòng bệnh gan thận mủ trên cá tra (Pangasianodon hypophthalmus)

This study aimed to evaluate the effect of dietary chamber bitter (Phyllanthus urinaria) extract on the resistance to Bacillary Necrosis in Pangasius (BNP) in striped catfish (Pangasianodon hypophthalmus). The experiment was in the completely randomized design with 3 triplicated treatments including 0% (control); 1% and 2% of P. urinaria extract. After 4 weeks of experimentation, fish were injected with Edwardsiella ictaluri. Samples were collected in the week 2nd and 4th of experiment and day 3rd of challenge test for immune parameter test. Fish mortalities among treatments were also recorded. The results showed that ecrythrocytes, leukocytes, monocytes, lymphocytes and lysozyme activity were significantly increased in supplemented P. urinaria treatments if compared those of the control (p

EVALUATION OF ANTI-ULCER ACTIVITY OF THE ETHANOLIC EXTRACT OF PHYLLANTHUS URINARIA IN EXPERIMENTAL ANIMALS

Objective: The objective of this study is to investigate the antiulcer activity of the ethanolic extract of Phyllanthus urinaria (EEPU). Methods: In vivo anti-ulcer activity of EEPU was evaluated in the present study at 500 mg/kg body weight by pyloric ligation, ethanol-induced ulcer, aspirin-induced ulcer, and cold restraint-induced ulcer model. The anti-ulcer activity was assessed by determining and comparing the gastric volume, pH, free and total acidity, ulcer number and its inhibition, and ulcer severity.Result: EEPU at 500 mg/kg body weight dose showed significant antiulcer activity by decreasing ulcer index, gastric volume, pH, and free and total acidity. Gastroprotective effect of EEPU was substantiated by histopathological studies of the stomach in ulcer and treated groups.Conclusion: It can be concluded from the results that EEPU has potential antiulcer activity.

Screening of Euphorbiaceae Plant Extracts for Anti-5α-reductase.

In our research program to find novel agents for alopecia from natural plant resources, we screened Euphorbiaceae plant extracts using an anti-5α-reductase assay. Among the samples tested, the extract of Phyllanthus urinaria showed the most potent activity with 24.3 and 64.6% inhibition at 50 and 200 µg/mL against the enzyme, respectively. The extract also suppressed the androgen activity of dihydrotestosterone in LNCaP cell line. These results show that the extract of P. urinaria may be a multi-potent agent for androgen-derived alopecia. We tested for activity on a hair regrowth model using mice. The extract of P. urinaria showed hair regrowth activity at 5 mg/mouse/d administration. Furthermore, the active principle for anti-5α-reductase activity was determined as stigmasterol glucoside from activity-guided fractionation and the IC50 was 27.2 µM. These results suggest that extract of P. urinaria may be a promising candidate anti-alopecia agent.

Corilagin inhibits breast cancer growth via reactive oxygen species-dependent apoptosis and autophagy.

Corilagin is a component of Phyllanthus urinaria extract and has been found of possessing anti‐inflammatory, anti‐oxidative, and anti‐tumour properties in clinic treatments. However, the underlying mechanisms in anti‐cancer particularly of its induction of cell death in human breast cancer remain undefined. Our research found that corilagin‐induced apoptotic and autophagic cell death depending on reactive oxygen species (ROS) in human breast cancer cell, and it occurred in human breast cancer cell (MCF‐7) only comparing with normal cells. The expression of procaspase‐8, procaspase‐3, PARP, Bcl‐2 and procaspase‐9 was down‐regulated while caspase‐8, cleaved PARP, caspase‐9 and Bax were up‐regulated after corilagin treatment, indicating apoptosis mediated by extrinsic and mitochondrial pathways occurred in MCF‐7 cell. Meanwhile, autophagy mediated by suppressing Akt/mTOR/p70S6K pathway was detected with an increase in autophagic vacuoles and LC3‐II conversion. More significantly, inhibition of autophagy by chloroquine diphosphate salt (CQ) remarkably enhanced apoptosis, while the caspase inhibitor z‐VAD‐fmk failed in affecting autophagy, suggesting that corilagin‐induced autophagy functioned as a survival mechanism in MCF‐7 cells. In addition, corilagin induced intracellular reactive oxygen species (ROS) generation, when reduced by ROS scavenger NAC, apoptosis and autophagy were both down‐regulated. Nevertheless, in SK‐BR3 cell which expressed RIP3, necroptosis inhibitor Nec‐1 could not alleviate cell death induced by corilagin, indicating necroptosis was not triggered. Subcutaneous tumour growth in nude mice was attenuated by corilagin, consisting with the results in vitro. These results imply that corilagin inhibits cancer cell proliferation through inducing apoptosis and autophagy which regulated by ROS release.

Hepatoprotective Effects of Corilagin Following Hemorrhagic Shock are Through Akt-Dependent Pathway.

ABSTRACTCorilagin, a component of Phyllanthus urinaria extract, possesses antioxidant, thrombolytic, antiatherogenic, and hepatoprotective properties, but the mechanism underlying these effects remains unclear. Previous studies showed that the Akt (protein kinase B) signaling pathway exerts anti-inflammatory and organ protective effects. The aim of this study was to investigate the mechanism of action of corilagin and determine whether these effects are mediated through the Akt-dependent pathway in a trauma-hemorrhagic shock-induced liver injury rodent model. Hemorrhagic shock was induced in male Sprague–Dawley rats; mean blood pressure was maintained at 35 mm Hg to 40 mm Hg for 90 min, followed by fluid resuscitation. During resuscitation, three doses of corilagin alone (1 mg/kg, 5 mg/kg, or 10 mg/kg, intravenously) were administered. Furthermore, a single dose of corilagin (5 mg/kg) with and without Wortmannin (1 mg/kg, PI3K inhibitor), Wortmannin alone, or vehicle was administered. Twenty-four hours after resuscitation, plasma alanine aminotransferase and aspartate aminotransferase concentration and hepatic parameters were measured. One-way ANOVA was used for statistical analysis. Hepatic myeloperoxidase activity and the concentrations of plasma alanine aminotransferase and aspartate aminotransferase, interleukin-6, tumor necrosis factor-α, intercellular adhesion molecule-1, and cytokine-induced neutrophil chemoattractant-1 (CINC-1) and CINC-3 increased following hemorrhagic shock. These parameters were significantly attenuated in corilagin-treated rats following hemorrhagic shock. Hepatic phospho-Akt expression was also higher in corilagin-treated rats than in vehicle-treated rats. The elevation of phospho-Akt was abolished by combined treatment with Wortmannin and corilagin. Our results suggest that corilagin exerts its protective effects on hemorrhagic shock-induced liver injury, at least, via the Akt-dependent pathway.

Preparation of ionic liquid-mediated imprinted monolith for selective capture and purification of corilagin.

A method for solid-phase extraction (SPE) of corilagin from natural plant extracts based on molecularly imprinted polymers (MIPs) was developed. For the preparation of corilagin-MIP monoliths, 4-vinylpyridine was used as functional monomer, and ethylene glycol dimethacrylate was used as cross-linking monomer, using a mixture of 1-butyl-3-methylimidazoliumtetrafluoroborate (ionic liquid)-N,N-dimethylformamide-dimethyl sulfoxide as a porogen. A morphological characteristic of the corilagin imprinted monolith was further studied by scanning electron microscopy and nitrogen sorption method. The greatest imprinting factor of COR was up to 9. The MIPs were used as solid-phase extraction (SPE) sorbents for purification of COR and the mean recoveries of corilagin was 78.0% with COR purity of 98.0% from the crude extract of phyllanthus urinaria L. The resulting COR-imprinted polymer also displayed the good performance of fragment imprinting polymer for gallic acid with the mean recoveries of 94.0% and purity of 99.7%.

Antiviral effects of Phyllanthus urinaria containing corilagin against human enterovirus 71 and Coxsackievirus A16 in vitro.

Human enterovirus 71 (EV71) and Coxsackievirus A16 (CA16) are major causative agents of hand, foot, and mouth disease (HFMD) especially in infants and children under 5 years of age. Despite recent outbreaks of HFMD, there are no approved therapeutics against EV71 and CA16 infection. Moreover, in a small percentage of cases, the disease progression can lead to serious complications of the central nervous system. In this study, we investigated the antiviral effect of corilagin and Phyllanthus urinaria extract, which contains corilagin as a major component, on EV71 and CA16 infection in vitro. Our results indicate that corilagin reduces the cytotoxicity induced by EV71 or CA16 on Vero cells with and IC50 value of 5.6 and 32.33 μg/mL, respectively. We confirmed the presence of corilagin in EtOAc and BuOH fractions from P. urinaria extract and this correlated with antiviral activity of the fractions against EV71 or CA16. Future studies will be required to confirm the antiviral activity of corilagin and P. urinaria extract in vivo. Challenging a model with a lethal dose of viral infection will be required to test this. Collectively, our work provides potential candidates for the development of novel drugs to treat HFMD.

A new flavone sulfonic acid from Phyllanthus urinaria

One new flavone derivative, urinariaflavone (1), and eight known compounds (2–9) were isolated from the methanolic extract of Phyllanthus urinaria L. leaves. Their structures were established by MS, 1D and 2D NMR experiments, and comparison with literature data. All isolated compounds were tested for their cytotoxic activity against CHO (Chinese hamster ovary) and J774 (Murine macrophage) cell lines. Cytotoxic effects (IC50s∼6.00–41.30μM) on both cell lines were observed for compounds 4, 5, and 7.

Two new acetylated flavonoid glycosides from Phyllanthus urinaria

Two new acetylated flavonoid glycosides, quercetin 3-O-α-l-(2,4-di-O-acetyl) rhamnopyranoside-7-O-α-l-rhamnopyranoside (1) and quercetin 3-O-α-l-(3,4-di-O-acetyl) rhamnopyranoside-7-O-α-l-rhamnopyranoside (2), together with two known compounds, quercetin (3) and quercetin 3-O-α-l-rhamnopyranoside (4), were isolated from the ethanol extract of Phyllanthus urinaria. The structures of the new compounds were determined on the basis of extensive spectroscopic data including IR, HR-ESI-MS, 1D NMR, and 2D NMR.

A novel green gelatin–agar microencapsulation system with P. urinaria as an improved anti-A. niger model

In this study, a novel green microencapsulation system was used to develop Phyllanthus urinaria (PU) extract containing microcapsules. Agar was used with gelatin as the wall matrix materials of microcapsules to prevent the use of toxic crosslinker formaldehyde. Microencapsulated PU extract was developed to improve the potential antifungal activities of PU water extracts. The active components and surface morphology of PU extract containing microcapsules were analyzed by liquid chromatography/mass spectrometry and scanning electron microscopy, respectively. The in vitro release study demonstrated that approximately 80% of drug was released after 120h. PU loaded microcapsules were shown to have a stronger anti-Aspergillus niger activity than the free drug.

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.

The relaxant effect of extract of Phyllanthus urinaria in the guinea-pig isolated trachea. Evidence for involvement of ATP-sensitive potassium channels.

This study analyses the relaxation induced by the hydroalcoholic extract of stems, leaves and roots from Phyllanthus urinaria (Euphorbiaceae) in the guinea-pig trachea (GPT) pre-contracted by carbachol. The hydroalcoholic extract of P. urinaria (0.1-10 mg mL-1) caused a graded relaxation in GPT with or without epithelium, with mean EC50 values of 1.94 (1.41-2.67) and 2.00 (1.47-2.78) mg mL-1 and Emax of 717 mg (+/- 16) and 627 mg (+/- 12), respectively. The relaxation in response to hydroalcoholic extract, like that to cromakalim (EC50 3.57 (2.75-4.64 microM) in GPT without epithelium, was fully abolished in the presence of high KCl concentrations (80 mM), and was significantly attenuated by tetraethylammonium (10 or 30 mM) or glibenclamide (0.1 or 3 microM). However, the relaxation caused by the hydroalcoholic extract was unaffected by apamin (0.1 or 1.0 microM), nitro-L-arginine (L-NOARG, 100 microM), methylene blue (10 microM) or by calcitonin gene-related peptide (CGRP) (8-37) (a CGRP antagonist, 0.1 microM). Both propranolol (1 or 3 microM) and [D-p-Cl-Phe6,Leu17]VIP (a vasoactive intestinal peptide (VIP) receptor antagonist, 0.1 microM) produced a significant displacement to the right (about 2-fold) of the relaxation response to hydroalcoholic extract of P. urinaria. Thus, the present results indicate that the ATP-activated potassium channels sensitive to glibenclamide, but not the small conductance calcium-activated potassium channels sensitive to apamin, largely contribute to the relaxation effect of the hydroalcoholic extract of P. urinaria in GPT. In addition, both beta 2 and VIP-mediated responses seem to account, at least in part, for the relaxation effect of the hydroalcoholic extract, as its relaxant response was partially attenuated by both propranolol and VIP receptor antagonist.