Phyllanthus Virgatus Research Articles

Development and Characterization of Copper Nanoparticle‐Embedded pH‐responsive Polymeric Microbeads for Drug Delivery and Antibacterial Applications

AbstractCopper nanoparticles have been utilized in a variety of therapeutic applications because of their inexpensive manufacturing and unique characteristics. In the current study, novel nanocomposites are developed to carry bioactive agents in a controlled manner. Copper nanoparticles (Cu‐NPs) were synthesized from copper sulfate using a leaf extract of Phyllanthus virgatus as a reducing agent. The obtained Cu‐NPs are loaded into the polymeric matrix of sodium alginate and locust bean gum to generate microbeads. The developed microbeads were characterized by Fourier‐transform infrared (FTIR), X‐ray diffraction (XRD), transmission electron microscopy (TEM), selected area electron diffraction (SAED), and scanning electron microscope (SEM). Swelling and in vitro release studies were performed at pH 2.0 and 7.4 at 37 °C. Metronidazole (MTD) was used as a model agent, and the antibacterial activity of microbeads was tested against Streptococcus mutans, Klebsiella pneumoniae, and Bacillus subtilis. The release kinetics and MTD release pattern were analyzed by employing the MTD release data into Korsmeyer‐Peppas equation and other kinetic models.

Germplasm conservation of Phyllanthus virgatus G. Forst. by encapsulation of In vitro derived leaf segments

The present study describes an efficient plant regeneration system for in vitro propagation of Phyllanthus virgatus through leaf explant. The leaf explants were cultured on Murashige & Skoog (MS) medium supplemented with BAP (1.5 mg L-1) and NAA (0.9 mg L-1 for efficient callus induction (82.26%). The maximum percentage of shoots (77.85%) was developed from BAP (2.0 mg L-1) and GA3 (0.5 mg L-1). The efficient rooting response (76.28%) was noted on a nutrient medium containing NAA (1.5 mg L-1) and IBA (0.2 mg L-1). Regenerated plantlets were successfully acclimatized and hardened off inside the culture room and then transferred to green house with a 72% survival rate.

A taxonomic revision of Phyllanthus subgenus Macraea (Phyllanthaceae)

Within the morphologically diverse pantropical genus Phyllanthus, many subgenera, sections and subsections are recognized. While most taxonomic revisions often focus on local floras, closely related and often resembling species are not always treated in full. Subgenus Macraea is here revised for the first time over its whole distribution, including an identification key and descriptions of its species with distributions, ecology, uses and vernacular names. The currently acknowledged varieties of Phyllanthus distichus are rejected due to inadequate morphological differences. Phyllanthus panayensis is synonymized with P. lancifolius. Phyllanthus alpestris has now become a variety of P. glaucophyllus because of the resemblance in morphology and distribution. The species complex around Phyllanthus virgatus remains taxonomically difficult, but Phyllanthus virgatus var. gardnerianus and Phyllanthus virgatus var. hirtellus are here recognized on the species level as P. gardnerianus, stat nov. and P. tararae, stat & nom. nov. A new species from the Philippines, Phyllanthus ridsdalei, is described.

Hepatoprotective effect of methanolic extracts of Phyllanthus virgatus against carbon tetrachloride-induced liver injury in rats

In this study, the methanolic extract of Phyllanthus virgatus was evaluated against carbon tetrachloride (CCl4) induced hepatotoxicity in rats. The results of this study indicated that Phyllanthus virgatus exhibited moderate protective effect at a dose of 100-200 mg/kg by lowering serum level of liver enzymes such as alanine amino transferees (ALT), glutamate pyruvate transaminases (SGPT), aspartate aminotransferase (AST), glutamate oxaloacetate transaminases (SGOT), and total protein to a significant extent. Further, no significant effects were seen on blood serum level at a dose of 100-200 mg/kg body weight. The highest activity was observed at a dose of 200 mg/kg with a reduction of serum concentration of ALT, AST, total bilirubin and total protein. The methanolic extract of P. virgatus showed significant decrease in the levels of liver enzymes, indicating the protection of hepatic cells thereby protecting against CCl4 induced hepatocellular injury.

Authenticity analyses of Phyllanthus amarus using barcoding coupled with HRM analysis to control its quality for medicinal plant product

The Phyllanthus genus, a plant used in traditional Thai medicine, has according to several pharmacopeias hepatoprotective properties. Not only is the anatomical morphology of these species relatively similar but they also share the Thai common names Look-Tai-Bai (ลูกใต้ใบ) and Yah-Tai-Bai (หญ้าใต้ใบ), which might cause confusion for laypersons. This study attempted to develop a method for accurate identification of Phyllanthus species, especially Phyllanthus amarus, and to detect contaminants in P. amarus products by using DNA barcoding coupled with high resolution melting (HRM) analysis (bar-HRM). Two plastid loci (rbcL and trnL) were chosen for DNA barcoding to generate a suitable primer for distinguishing Phyllanthus species by HRM analysis. The five species of Phyllanthus were subjected to amplification for testing the specificity and discrimination power of the designed primers derived from rbcL and trnL regions. Sensitivity of the method (DNA barcoding conjugated with HRM) to detect adulterant in P. amarus samples was evaluated. The commercial P. amarus products obtained from a local market were authenticated. The primer pair derived from trnL DNA barcoding (PhylltrnL) had more specificity and power of discrimination for Phyllanthus species than that derived from rbcL DNA barcoding (PhyllrbcL). The result showed that Tm of P. amarus, Phyllanthus urinaria, Phyllanthus debilis, Phyllanthus airy-shawii, and Phyllanthus virgatus was 74.3±0.08, 73.04±0.07, 73.36±0.05, 72.21±0.06, 72.77±0.15°C, respectively. This method proved to be a very sensitive tool that can be used for rapid detection of contamination as low as 1% of other Phyllanthus species in P. amarus admixtures. All commercial products of P. amarus obtained from a local market in Thailand were found to contain pure raw materials of P. amarus without any substitution or contamination. Our results indicated that the use of DNA barcoding coupled with HRM was an efficient molecular tool for correct species identification. This molecular tool provides a noteworthy benefit for quality control of medicinal plants and industry plants for pharmacological prospects.

Alleviation of hyperglycemia and hyperlipidemia by Phyllanthus virgatus forst extract and its partially purified fraction in streptozotocin induced diabetic rats.

Since, we previously demonstrated that sequentially extracted methanolic fraction showed marked antioxidant and antidiabetic property in vitro, the present study was design to evaluate the beneficial effects of Phyllanthus virgatus methanolic extract and its partially purified fraction on hyperglycemia and hyperlipidemia in streptozotocin (STZ) induced diabetic rats. The plant extract was subjected to repeated thin layer chromatographic fractionation followed by GC-MS analysis of active fraction. TLC data illustrated the presence of six prominent bands and the prelimnary screening of these bands against α-amylase inhibitory activity showed that the band with Rf value 0.514 has marked inhibitory property (IC50, 48 µg/ml). The diabetic rats were treated for four weeks with methanolic extract of P. virgatus (50 and 10 mg/rat/day), partially isolated active fraction (0.5 and 0.1 mg/rat/day) and glibenclamide (0.1 mg/rat/day). The level of fasting blood glucose (FBG), hemoglobin, glycated hemoglobin (HbA1c) and insulin were significantly alleviated in plant extract and partially purified fraction treated group after 28 days of administration. Moreover, total cholesterol (TC), triglycerides (TG), low density lipoprotein-cholesterol (LDL-C), very low density lipoprotein-cholesterol (VLDL-C) and high density lipoprotein cholesterol (HDL-C) were also markedly ameliorated in the entire treatment group, with a maximum restoration observed in group treated with partially purified fraction (0.5 mg/rat/day). The results demonstrate a strong antidiabetic and hypolipidemic impact of plant extract and its partially purified fraction coupled with their potent antioxidative property, which can provide additional benefits in the inhibition of oxidative stress and hence in the prevention and treatment of diabetes as well as diabetes linked hyperlipidemia.

Comparison of the Antioxidant and Cytotoxic Activities of Phyllanthus virgatus and Phyllanthus amarus Extracts

Objective: To determine the antioxidant activity and cytotoxicity of Phyllanthus virgatus crude extract compared to Phyllanthus amarus.Methods: Phenolic contents of the hydromethanolic extracts were measured using Folin-Ciocalteu reagent. Antioxidant activity was evaluated by the 2,2-diphenyl-1-picrylhydrazyl hydrate free radical scavenging and antilipid peroxidation assays. Cytotoxicity on human hepatoma HepG2 cells was assessed by trypan blue and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide viability assays. A stereomicroscope was used to observe and photograph the morphology of the cells. Oxygen consumption of the HepG2 cells was measured using a Clark oxygen electrode. Results: The extract of P. virgatus, which contained more phenolic compounds than P. amarus extract, had higher cytotoxicity and showed higher free radical scavenging activity and more inhibition of peroxidation in a linoleic acid system. P. virgatus extract conspicuously increased the oxygen consumption of HepG2 cells, while P. amarus extract had little stimulatory effect. Conclusion: The hydromethanolic extract of P. virgatus had stronger antioxidant and cytotoxic action than P. amarus extract. The stimulation of HepG2 cell respiration by P. virgatus extract suggests the extract alters mitochondrial function; this action could play a role in the cytotoxicity of this plant.

Cell death and impairment of mitochondrial functions induced by Phyllanthus virgatus: a comparison study

Cell death and impairment of mitochondrial functions induced by Phyllanthus virgatus: a comparison study

Protective effect of Phyllanthus virgatus extract on Cd 2+ ‐induced loss of mitochondrial membrane potential

Phyllanthus virgatus, family Euphorbiaceae, is a plant traditionally employed for the treatment of several diseases in Thailand, especially for liver ailments. The mechanisms responsible for the hepatoprotective action of P. virgatus, however, are not well defined. In this study, the protection of P. virgatus extract on mitochondrial dysfunction induced by Cd2+ was examined with isolated rat liver mitochondria. It was found that the extract of P. virgatus at 80 to 240 μg/ml blocked the collapse of mitochondrial transmembrane potential induced by 5 μM CdCl2 in a dose-dependent manner. Similar results were also obtained with Ca2+-induced loss of mitochondrial membrane potential. Increasing the cation concentrations did not diminish this protective effect of the extract. The extract also inhibited Cd2+-induced cytochrome c release from mitochondria. Cd2+ has been suggested to cause mitochondrial damage by opening the permeability transition pore (PTP) after entering the mitochondria via Ca2+ uniporter. We therefore postulated that P. virgatus extract may act by blocking Ca2+ uniporter to impede Cd2+ influx into the mitochondria and/or by inhibiting the Cd2+-induced PTP opening.

In vitro antioxidant activities of methanol extracts of five Phyllanthus species from India

In this study, the antioxidant activity of methanol extracts of five plants from the genus Phyllanthus was evaluated by various antioxidant assays, including total antioxidant, free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, nitric oxide scavenging, reducing power and metal ion chelating activities. The various antioxidant activities were compared to standard antioxidants such as butylated hydroxytoluene and ascorbic acid. All the extracts showed strong antioxidant activity in all the tested methods. Among the five plants Phyllanthus debilis has been found to possess the highest activity in all tested models, the activity decreased in the order Phyllanthus debilis> Phyllanthus urinaria> Phyllanthus virgatus> Phyllanthus maderaspatensis> Phyllanthus amarus. In addition to the antioxidant activity of these plants, the total phenolic compounds, flavonoids and flavonols were measured in the extracts. A correlation between the antioxidant activity and total phenolic content was observed.

Heavy Metal and Nutrient Concentration in Soil and Plants Growing on a Metalliferous Chromite Minespoil

Metal contamination in soil and plant samples from a chromite mine and its adjoining regions was determined. The metal concentration varied in stem, leaf and root of different tree species. In the case of shrubs, the highest concentration of iron (18.5 mg kg−1) was detected in the stem of Combretum roxburghii. The concentration of aluminium varied from 1.8 - 5.3 mg kg−1 dry weight, whereas the nickel content was found to be the highest in the stem of Calotropis gigantea. In the case of herbs, chromium concentration was highest (60.9 mg kg−1 dry weight) in Evolvulus alsenoides and the lowest (18.8 mg kg−1 dry weight) in Andrographis paniculata. There was a significant correlation observed between chromium in soil with the root of tree species like Lagerstroemia parviflora, Madhuca longifolia, Anogeissus latifolia and Haldina cordyfolia. Nickel in soil was significantly correlated with the stem and leaf of all the tree species except Chloroxylon sweitenia. Iron in soil showed correlation with the stem and leaf of Chloroxylon sweitenia. Among the shrubs (Calotropis gigantea, Combretum roxburghii and Smilax zeylanica), chromium in soil showed a correlation with the root. Nickel in soil was positively correlated with the stem and leaf of Calotropis gigantea and Combretum roxburghii. Among the herbs, chromium in the whole plant of Evolvulus alsenoids, Solanum surattense and Phyllanthus fraternus showed significant positive correlation with soil; nickel in Solanum surattense showed significant positive correlation with soil. The positive correlation coefficient was observed between iron in the whole plant and soil on Phyllanthus virgatus, Phyllanthus fraternus and Andrographis paniculata. The above information would be useful for the establishment of a vegetation cover on the minewaste heaps.

First total synthesis of tetrasubstituted tetrahydrofuran lignan, (−)-virgatusin

The first asymmetric total synthesis of (−)-virgatusin, a new furanolignan, isolated from phyllanthus virgatus, was accomplished in a stereoselective manner by nucleophilic addition of organolithium reagent to the functionalized lactone elaborated from dihydroxyacetone dimer followed by asymmetric deoxygenation of the hemiketal intermediate.

Tannins, flavonol sulfonates, and a norlignan from phyllanthus virgatus

Investigation of the constituents of Phyllanthus virgatus has led to the isolation of five new compounds, including a norlignan, 2-(3, 4-methylenedioxybenzyl)-4-(3,4-methylenedioxyphenyl)-3-butyne-1, 2-diol named virgatyne (1); a hydrolyzable tannin, virganin (2); and three flavonoid sulfonates, galangin-8-sulfonate (4), galangin-3-O-beta-D-glucoside-8-sulfonate (5), and kaempferol-8-sulfonate (6). Their structures were established by spectral and chemical methods.

A New Lignan from Phyllanthus virgatus

A new lignan, (−)-7,8-cis-8,8‘-trans-7‘,8‘-trans-7-(3,4-(methylenedioxy)phenyl)-7‘-(3‘,4‘-dimethoxyphenyl)-8,8‘-bis(methoxymethyl)tetrahydrofuran, named virgatusin (1), was isolated from the EtOH extract of the whole herb of Phyllanthus virgatus, together with seven known lignans and indole-3-carboxylic acid.