Amarus Extract Research Articles

Effects of Polyphenols and Lignans of Phyllanthus amarus Schumach. and Thonn. on IL-1β and TNF-α Secretions from LPS-induced THP-1-derived Macrophages

Background: Phyllanthus amarus exhibited immunosuppressive and anti-inflammatory effects in several in vitro and in vivo studies. However, there is no report on the inhibitory effects of its secondary metabolites on pro-inflammatory cytokines secretion from human THP-1-derived macrophages. background: Phyllanthus amarus exhibited immunosuppressive and anti-inflammatory effects in several in vitro and in vivo studies. However, there is no report on the inhibitory effects of its secondary metabolites on pro-inflammatory cytokines secretion from human THP-1-derived macrophages. Objective: The objective of this study was to correlate the polyphenols (ellagic acid (EA), gallic acid (GA), geraniin (GER), and corilagin (COR)) and lignans ((phyllanthin (PHY), hypophyllanthin (HYPO), niranthin (NIR), phyltetralin (PHYLT), and isolintetralin (ISO)) of 80% ethanol extract of P. amarus with their inhibitory effect against IL-1β and TNF-α secretions from LPS-induced THP-1- derived macrophages. Methods: Chemical profiling of P. amarus was carried out by LC-MS/MS analysis. Validated qualitative and quantitative reversed-phase HPLC analyses of the P. amarus extract were performed for the determination of lignan and polyphenol contents. Human THP-1-derived macrophages were prepared by treatment of THP-1 cells with phorbol 12-myristate 13-acetate (PMA). The inhibition of cytokines released by the extract, lignans and polyphenols in the cells was investigated using ELISA assay. method: Chemical profiling of P. amarus was carried out by LC-MS/MS analysis. Validated qualitative and quantitative reversed phase HPLC analyses of the P. amarus extract were performed for determination of lignan and polyphenol contents. Human THP-1 derived macrophages were prepared by treatment of THP-1 cells with phorbol 12-myristate 13-acetate (PMA). The inhibition of cytokines release by the extract, lignans and polyphenols in the cells were investigated using ELISA assay. Results: P. amarus extract and its chemical constituents significantly reduced the levels of IL-1β and TNF-α in a dose-dependent manner. At a dose of 50 μM, COR exhibited a maximum inhibition of 81.11% on TNF-α secretion, while GER showed 72.56% inhibition on IL-1β secretion. COR demonstrated the strongest inhibition of TNF-α secretion, exhibiting an IC50 value of 9.06 μM, which was comparable to that of dexamethasone (7.07 μM). Meanwhile, GER was the most potent against IL- 1β secretion, exhibiting an IC50 value of 20.09 μM. In the case of TNF-α secretion, the order of potency observed among the active compounds, with regard to IC50 value, was COR > GER > HYPO > PHY > NIR > GA > EA >ISO > PHYLT. For IL-1β secretion, the order of potency was GER > NIR > COR > GA > EA > PHY > HYPO > PHYLT > 1SO. Conclusion: The polyphenol contents of P. amarus, especially COR and GER, contributed significantly to the suppression of cytokines secretion, and they have the potential to be developed into agents for the treatment of pathologic inflammation.

Stimulatory Effects of Phyllanthus amarus Extract on the Growth Performance, Hematobiochemical Activity, Antioxidative Status and Immune Response of Clarias gariepinus

This study investigated the antibacterial activity of Phyllanthus amarus extracts and its influence on the performance of Clarias gariepinus fingerlings. The fish were fed diets containing 0.0, 0.5, 1.0, 1.5 or 2.0 g P. amarus methanol extract (PAE) / kg basal diet for 84 days. Thereafter, blood samples were collected for hematological and biochemical analyses. At the end of the 84-day feeding trial, fish were challenged with Aeromonas hydrophila, after which immune response parameters were measured. The data obtained were analyzed using one-way analysis of variance at p<0.05. The results showed significant antibacterial activity of PAE against A. hydrophila, and its application at 0.5-1.5 g significantly promoted growth performance, with the highest at 1.0 g. Hematocrit, hemoglobin and lymphocytes were enhanced at 0.5-1.5 g PAE. All the fishes fed PAE-supplemented diets had lower concentrations of serum liver enzymes; while the values of creatinine, glucose and total bilirubin did not differ among the treatments. Glutathione peroxidase, glutathione‐S‐transferase, lysozyme, phagocytic and respiratory burst activities increased in all PAE-fortified treatments, with highest fish survival in 1.0 g PAE treatment. Therefore, the inclusion of 1.0 g Phyllanthus amarus extract is recommended as a dietary supplement for Clarias gariepinus.

Effects of Psidium guajava and Phyllanthus amarus extracts on digestive enzyme activity and growth of Pangasianodon hypophthalmus fingerlings under high-temperature stress

The rise in water temperature by global warming is of high concern to aquaculturists. In this study, the effects of extracts-based diets on digestive enzymes, and growth performance in Pangasianodon hypophthalmus fingerlings under elevated temperatures were investigated. Four distinct diets (control, Psidium guajava, (0.2%/kg) – Pg0.2, Phyllanthus amarus (0.5%/kg) – Pa0.5, and a mixture of Pg0.2 and Pa0.5 - Mix.) were administered to fish fingerlings for 42 days, followed by 4 days of temperature elevation. Fish were then continuously subjected for 42 days to temperatures of 27, 31, and 35°C to evaluate enzymatic activities and growth performance of fish. The results showed that although there is no interaction between two experimental factors on digestive enzyme activity and growth performance of fish, Pg0.2 followed by Mix groups accelerated digestive enzymes (trypsin, chymotrypsin, amylases and pepsin). Besides, enzymatic activities increased from 31°C to 35°C. The highest growth was observed from fish at 35°C followed by those at 31°C (p<0.05) which was significantly higher than the control (27°C); however, there was no significant difference in survival rate. In conclusion, these findings suggested 2 appropriate diets (Pg0.2 and Mix) for optimizing growth of this species and consequently contributing to the sustainable aquaculture under the global warming scenario.

Antibacterial properties of Allium sativum against Ornithobacterium rhinotracheale and Phyllanthus amarus extracts against Escherichia coli

Antibacterial activity of garlic extracts (Allium sativum) against Ornithobacterium rhinotracheale (ORT) and Phyllanthus amarus extracts against Escherichia coli (E. coli) were evaluated by agar diffusion assay. The results showed that ORT were sensitive to pure garlic extract and garlic extract from alcohol. The diluted extract inhibited ORT at moderate levels. Relating to E. coli, 250 mg/mL of Phyllanthus extract destroy the bacteria while minimal bactericidal concentration of doxycycline and amoxicillin against Escherichia coli were 16 μg/mL and 64 μg/mL, respectively. The results contribute to evaluate garlic extract and Phyllanthus extract as antibiotic alternatives in animal production.

Keap1/Nrf2-independent antioxidative activity of Phyllanthus amarus extract in zebrafish

The Keap1 protein (Kelch-like ECH-related protein 1) and the Nrf2 transcription factor (NF-E2-related factor 2) are important systems for maintaining homeostasis, redox, and metabolism. Based on the Keap1/Nrf2 pathway, the antioxidative mechanism of P. amarus extract (PAE) was predicted. In this paper, we evaluated the protective effects of PAE on the oxidative toxicity induced by sodium arsenite (NaAsO2) and hydrogen peroxide (H2O2) in zebrafish larvae. We first determined that the LC50 values for NaAsO2, H2O2, and PAE at 3.5 days postfertilization (dpf) were 1 mM, 3 mM, and 200 μg/mL, respectively. Then, to assess the antioxidant effects of P. amarus, 3.5 dpf zebrafish larvae were pretreated with PAE at concentrations of 0, 50, 75, and 100 μg/ml for 12 h, and then the PAE solution was replaced with 1 mM NaAsO2 or 3 mM H2O2 to assess challenge survival within 48 h. Interestingly, all three concentrations, 50, 75, and 100 μg/mL PAE, increased the survival rate of zebrafish larvae compared to those larvae exposed to only 1 mM NaAsO2. Similarly, PAE at concentrations of 75 and 100 μg/mL protected zebrafish larvae after exposure to 3 mM H2O2. Real-time qPCR analysis was performed after 3.5 dpf zebrafish larvae were exposed to 100 μg/mL PAE for 12 h to verify whether the increasing antioxidative activity is depended on the Nrf2 pathway. The expression of the Nrf2 target genes glutathione-S-transferase Pi 1 (gstp1) and peroxidase 1 (prdx1) was assessed using real-time qPCR. However, the expression of this gene was not significantly different between control larvae and PAE-treated larvae. Thus, PAE induces antioxidant activity in zebrafish in a Keap1/Nrf2-independent manner.

Antibacterial Activity of <i>Phyllanthus</i><i> </i><i>Amarus</i> (Schum and Thonn) Extract Against <i>Salmonella </i><i>Typhi</i> Causative Agent of Typhoid Fever

The study was conducted to assess the antibacterial activity of Phyllanthus amarus (Schum and Thonn) extract against Salmonella typhi causative agent of typhoid fever at the laboratories of the Departments of Chemistry and Theoretical and Applied Biology of the College of Science, Kwame Nkrumah University of Science and Technology, Kumasi. The objectives were to determine the highest yield of crude extract of P. amarus using different proportions of water to ethanol and to determine the sensitivity of Salmonella typhi to these. Three different extraction procedures were carried out. In the first procedure, seven extraction setups each containing different proportions of the two extract (water and ethanol) were used with 10g of the plant sample. In the second procedure, eight setups were used for the two solvents. Ten grams of both fresh and dry plant sample were extracted in two different 200ml of water and in another two different 200ml of water; 20g of both fresh and dry plant sample were again extracted. The same procedure was repeated using ethanol as the solvent. In the third procedure, 10g each of fresh plant sample were boiled in 100ml and 200ml of water for 30 minutes. A sensitivity test to determine the zones of inhibition for the various plant extracts was done on Salmonella typhi isolated from human. Results from the crude yield of P. amarus using water only had the highest crude yield of 2.57g, followed by ethanol only which was 2.52g. The sensitivity studies conducted on the fresh P. amarus indicated that aqueous extract of P. amarus inhibited S. typhi to a zone of 5.00mm in 10g/200ml and 7.17mm in 20g/200ml. Ethanol extract also recorded an inhibition zone of 2.67mm and 5.33mm in 10g/200ml and 20g/200ml respectively. Again, sensitivity studies using dry P. amarus samples showed that the aqueous extracts recorded a zone of inhibition of 7.33mm in 10g/200ml and 13.50mm in 20g/200ml. Also ethanol extracts also recorded an inhibition zone of 6.83mm in 10g/200ml and 10.50mm in 20g/200ml. Significant differences were observed among the extracts and the control in both 10g/200ml and 20g/200ml concentrations (P<0.05). Aqueous and ethanol extracts of P. amarus proved inhibitory to S. typhi.

An insight into the potent medicinal plant Phyllanthus amarus Schum. and Thonn.

The online version contains supplementary material available at 10.1007/s13237-022-00409-z.

Effect of Phyllanthus amarus in sodium arsenite-induced tissue damage

This study aims to evaluate the effect of alcoholic extract of Phyllanthus amarus leaves on arsenic-induced histological changes in Wistar rats. Rats were divided into 4 groups: Group I: normal control; Group II: rats received sodium arsenite (40 mg/kg); Group III: rats received sodium arsenite (40 mg/kg) + P. amarus extract (100 mg/kg); Group IV: rats received sodium arsenite (40 mg/kg) + extract (200 mg/kg). All groups were treated by oral gavage with sodium arsenite for 28 days and the animals were subsequently administered (oral) with 100 and 200 mg/kg extract, once daily for two weeks. Animals were sacrificed 24 hours after the last treatment and different organs were collected for histopathological analysis. Results revealed mild to severe type of necrosis and degenerative changes in brain, kidney, and liver of arsenic fed animals. In rats administered with extract showed significant improvements, and the normal histological feature of the cells was almost restored in rats. These suggest that P. amarus can be used to treat arsenic-induced multi-organ damages.

Bioactivity of Leaf Extract of Phyllantus amarus against Fungal Pathogens Associated with Sweet Orange Rot Disease

This study was carried out to determine disease incidence and control of fungal pathogens associated with rot of sweet orange using Phyllantus amarus leaf extract. Forty sweet oranges (Citrus sinensis var. Valencia) were collected from the markets and taken to the laboratory. Isolation and identification of the fungi were carried out using standard procedures and the efficacy of the leaf extracts of Phyllantus amarus was determined on the isolated fungi. The result of this study showed that four pathogens (Aspergillus niger, Aspergillus flavus, Penicillum expansum and Rhizopus stolonifer) were identified to be associated with the rot. Rot incidence was upto of 35%. Pathogenicity confirmed the characteristic features similar to the original diseased samples. The effect of the leaf extract of Phyllantus amarus on the isolated fungi shows that the rate of inhibition increases with increase in concentration of the P. amarus extract, The P. amarus extract was effective against A. flavus had the highest zone of inhibition (10.667). The inhibition zone was lesser for R. stolonifer among the pathogens (8.333). Sweet orange should be consumed after harvest or can be stored in refrigerator for period of 1=2 weeks. The fruit should be discarded if any alteration in color or taste of the fruit is noticed as this can be hazardous to human health.

Phyllanthus amarus Schumach. & Thonn. and Momordica charantia L extracts improve memory function, attenuate cholinergic and purinergic dysfunction, and suppress oxidative stress in the brain of doxorubicin–treated rats

BackgroundPhyllanthus amarus (P.A) and Momordica charantia (M.C) are common herbs known for their various pharmacological importance. However, limited studies have been done regarding their neuroactive effect. This study investigates the role of Phyllanthus amarus (P.A) and Momordica charantia (M.C) extracts on cognitive behavior and some neurochemicals associated with memory function in rats administered with doxorubicin (DOX). MethodologyNormal rats were pre-treated with P.A and M.C (200 and 400 mg/kg bwt.) for 14 days, while on the last day, DOX (15 mg/kg bwt. i.p.) was administered in a single dose. The cognitive behavior was evaluated using Y-maze and Morris Water Maze Tests. Thereafter, the biochemical assays [acetylcholinesterase (AChE), butyrylcholinesterase (BChE), arginase, 5′- nucleotidase, adenosine deaminase (ADA), angiotensin-I converting enzyme (ACE), and catalase activities, thiobarbituric acid reactive species, and non-protein thiol levels] were determined in rats’ brains. The extracts were characterized using a high-performance liquid chromatography-diode array detector (HPLC-DAD). ResultP.A and M.C extract restored cognitive behavior, nootropic-related enzyme activities, and improved antioxidant biomarkers altered by DOX. Furthermore, phenolic characterization of the extracts revealed gallic, chlorogenic, caffeic, and ellagic acids, catechin, epicatechin, rutin, quercitrin, isoquercitrin, quercetin, and kaempferol. Molecular docking analysis revealed that all the phenolic compounds detected in the studied plants had binding affinities for ACE, AChE, 5′-nucleotidase, and ADA proteins as the target proteins. ConclusionExtract from P.A and M.C effectively suppressed DOX-induced neurotoxicity by improving brain antioxidant status. Thus, they could play a significant role in restoring cognitive function altered by DOX treatment.

The effect of the chamber bitter (Phyllanthus amarus) extract on the quality of the snakehead (Channa striata) fillets during ice storage

The effect of the chamber bitter (Phyllanthus amarus) extract on the quality of the snakehead (Channa striata) fillets during ice storage

Antimicrobial activity of Sida acuta, Phyllanthus amarus and Phyllanthus muellerianus against microorganisms implicated in urinary tract infections

Increasing level of antimicrobial resistance among bacterial pathogens causing Urinary Tract Infection (UTI) is one of the most significant public health challenges globally. Hence, the search for alternatives from medicinal plants. This study investigated the efficacy of Phyllanthus amarus (PA), Phyllanthus muellerianus (PM) and Sida acuta (SA) leaf extracts on microorganisms implicated in UTI. Mid-stream urine samples collected from 100 patients clinically diagnosed with UTI were cultured. The microorganisms isolated were identified using their morphological and biochemical characteristics. Methanol leaf extracts of the three plants were obtained by cold maceration in 60% methanol. Crude extract of PM was thereafter purified by solvent partitioning. Antibiotic susceptibility test was determined using the Kirby Bauer disc diffusion. Antimicrobial effects of the extracts and oil was ascertained using agar well diffusion. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentrations (MBC) were also determined. Rate of kill and mechanism of action of the purified extract of PM on isolates were investigated. Cytotoxicity of plant extracts were assayed on brine shrimps while synergism of the purified extract with ciprofloxacin was ascertained using overlay inoculum susceptibility disc method. Antioxidant and phytochemical analyses of the extracts were conducted using standard methods. Phytochemical analysis of the leaf extracts showed the presence of alkaloids, tannins, saponins and steroids. Antioxidant assay also indicated SA had the highest total flavonoids and phenol content of 339.86 mgQUE/g and 27.63 mgGAE/g. Microorganisms isolated include: Escherichia coli (24%), Proteus mirabilis (24%), Staphylococcus aureus (19%), Klebsiella pneumoniae (13%), Candida albicans (11%), Enterobacter sp. (5%) and Citrobacter sp. (4%). The crude extract of PA had zone of inhibition ranging from 16.7 ± 1.53 mm to 24 ± 1.00 mm while SA crude extract had 14.7 ± 1.53 mm to 27 ± 2.00 mm. PM crude extract had inhibition zones of 17 ± 1.00 mm to 22.3 ± 2.12 mm. The MIC and MBC ranged from 6.25 mg/ml to 50 mg/ml and 12.5 mg/ml to 50 mg/ml respectively. Ethyl acetate fraction of PM showed the highest percentage yield and had a zone diameter range from 13.5 ± 1.00 mm to 28 ± 1.53 mm with MIC and MBC ranges of 6.25 mg/ml – 12.5 mg/ml and 25 mg/ml to 50 mg/ml respectively. Synergism with ciprofloxacin was observed at 25% of the microorganisms, 50% antagonism and 25% additively. Toxicity analysis showed lethal dose concentrations of 19.05 mg/ml, 25.12 mg/ml and 130.11 mg/ml for PM, PA and SA respectively. The findings of this study suggest that the methanol extracts of the medicinal plants used in this study does possess a potent lead molecule in combating microorganisms causing UTI.
 Key words: Antimicrobial activity, Phyllanthus muellerianus, Phytochemicals, Toxicity, UTI,

Effects of Phyllanthus amarus and Euphorbia hirta Dip Treatments on the Protection of Striped Catfish (Pangasianodon hypophthalmus) Fillets against Spoilage during Ice Storage

ABSTRACT This study was conducted to evaluate the effects of herbal extracts on the quality of striped catfish (Pangasianodon hypophthalmus) fillets throughout the storage period. Quality changes during ice storage of striped catfish fillets were studied after dip treatments in aqueous solutions of ethanolic extract of Phyllanthus amarus (0.02% and 0.04%, w/v) or Euphorbia hirta (0.06% and 0.02%, w/v). The control (dipped in tap water) and the treated fish samples were analyzed periodically for total viable counts (TVC), peroxide value, physicochemical parameters (pH, texture), and sensory properties. Results indicated that Pseudomonas spp. and Listeria monocytogenes were absent in the raw fish fillets, reflecting the safety of raw materials regarding specific spoilage and psychrotrophic pathogenic microorganisms. Fish fillets dipped in 0.04% P. amarus extract solution displayed lower TVC values (4.76 log10 CFU/g) during the initial storage period compared to treatment with E. hirta extracts. Dip treatments in water containing P. amarus or E. hirta extracts significantly reduced the primary lipid oxidation in fish samples. In conclusion, the dip treatment in water containing 0.04% P. amarus or 0.06% E. hirta extract was effective to maintain a good sensory quality of the fish fillets and to prolong their shelf life up to 8 days under ice storage.

Camellia sinensis and Phyllanthus amarus Ethanol Extracts Induced Apoptosis and Cell Cycle Arrest on Human Leukemic Cell Lines

Leukaemia is a heterogeneous hematologic malignancy characterized by unregulated proliferation of the early blood-forming cells which starts in bone marrow. The basic strategy of leukaemia therapy involves the induction of leukemic cells apoptosis. Research on natural products have shown that some plant derivatives have anticancer properties by inducing apoptosis of leukemic cells. Plants such as Camellia sinensis and Phyllanthus amarus are those that had gained a wide interest due to their anti-cancer effect. The aim of this study was to investigate the anti-cancer effects of C. sinensis and P. amarus extracts on human leukemic cell lines by analysing the cell cycle and determining the apoptotic state. The cell lines were treated with ethanolic plant extracts at the concentrations of 31.25 - 500 µg/mL for 24 h followed by MTT assay to determine the IC50. The IC50 of C. sinensis and P. amarus on the U937 cells were 170±10.39 and 210±6.78 µg/mL, respectively. Flow cytometric analysis of apoptosis using Annexin V/propidium iodide (PI) staining was also performed. C. sinensis extract at 170 µg/mL significantly increase apoptosis in U-937 (p<0.001), Jurkat (p<0.05) and K-562 cells (p<0.01) when compared to untreated cells. Meanwhile, P. amarus extract at 210 µg/mL significantly induced apoptosis in both U937 and K562 cells (p<0.05) but not Jurkat cells and caused cell cycle arrest at S phase in U-937 cells (p<0.001) and at G0/G1 in K652 cells (p<0.05) when compared to control. Based on the findings, both C. sinensis and P. amarus extracts showed potential in inducing apoptosis in human leukemic cell lines. In addition, P. amarus has the capability to disrupt cell cycle.

Phenolic profiles and biological properties of traditional Phyllanthus amarus aqueous extracts used for diabetes

• Traditional infusion and decoction Guadeloupe’s methods were used in this study. • Caffeoylhexaric acid has been detected, for the first time, in P. amarus species. • All extracts show strong antioxidant activity and glucose-6-phosphatase inhibition. • Dried plant extracts exhibit strong rise of glucose uptake similar to insulin. For the first time, a complete study, including the fresh and dried aerial parts of Phyllanthus amarus was performed using Guadeloupe's population traditional extraction methods. In this study, the antidiabetic and antioxidant activities of P. amarus infusion and decoction in water with short boiling time, were evaluated using glucose-6-phosphatase enzyme in H4IIE hepatic cells inhibition and stimulation of glucose uptake in C2C12 skeletal muscle cells, as well as using 2,2-diphenyl-1picrylhydrazyl (DPPH) radical scavenging activity and ferric reducing antioxidant power assays. A compound, caffeoyl hexaric acid, was tentatively identified for the first time in P. amarus extracts, using UHPLC-HRMS. Only dried aerial parts extracts exhibited strong stimulation of glucose uptake similar to that of insulin with an increase of 23%. The infusion of fresh material appeared to be the best way for aerial parts maximal anti-hyperglycemic activity with 55.8 ± 2.61% of glucose-6-phosphatase inhibition.

In Vitro Anti-Leptospiral Activity of Phyllanthus amarus Extracts and Their Combinations with Antibiotics.

Despite modern medicine, there is an increasing trend for cases of the bacterial infection leptospirosis, and this has led to the exploration of alternative medicines from various sources including plants. The aim of this study was to investigate the in vitro anti-leptospiral activity of Phyllanthus amarus extracts alone and combined with penicillin G, ceftriaxone, and doxycycline. Antimicrobial susceptibility testing was performed using the microdilution broth technique upon methanol extract (ME), aqueous extract (AE), and antibiotics against the Leptospira interrogans serovars Australis, Bataviae, Canicola, and Javanica, to determine minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs). The results were analyzed using an ELISA microplate reader combined with microscopic analysis. Synergy testing using a checkerboard assay was performed to determine the fractional inhibitory concentration index values of extracts combined with antibiotics against leptospires. Scanning electron microscopy (SEM) was used to investigate morphological changes of leptospires caused by potential anti-leptospiral agents alone and combined with antibiotics. The MICs and MBCs for P. amarus extracts ranged from 100 to 400 µg/mL for AEs and from 400 to 800 µg/mL for MEs. Penicillin G was the most effective anti-leptospiral drug, with MICs and MBCs ranging from <0.01 to 0.78 and <0.01 to 3.13 µg/mL, respectively, followed by ceftriaxone, with both MICs and MBCs ranging from 0.05 to 0.78 µg/mL, and doxycycline, with MICs and MBCs ranging from 0.39 to 3.13 µg/mL and 12.5 to 25 µg/mL, respectively. Combinations of P. amarus extracts and antibiotics did not show synergistic effects on all tested Leptospira serovars, with some combinations demonstrating antagonistic effects. SEM analysis, however, showed distorted Leptospira surfaces. P. amarus AE performed better anti-leptospiral activity than P. amarus ME. The morphological effects of P. amarus extract alone and its combination with antibiotic on Leptospira cells revealed promising anti-leptospiral properties.

Mechanistic Studies of the Antiallergic Activity of Phyllanthus amarus Schum. & Thonn. and Its Compounds.

Phyllanthus amarus Schum. & Thonn. (Phyllanthaceae) is a medicinal plant that is commonly used to treat diseases such as asthma, diabetes, and anemia. This study aimed to examine the antiallergic activity of P. amarus extract and its compounds. The antiallergic activity was determined by measuring the concentration of allergy markers release from rat basophilic leukemia (RBL-2H3) cells with ketotifen fumarate as the positive control. As a result, P. amarus did not stabilize mast cell degranulation but exhibited antihistamine activity. The antihistamine activity was evaluated by conducting a competition radioligand binding assay on the histamine 1 receptor (H1R). Four compounds were identified from the high performance liquid chromatography (HPLC) analysis which were phyllanthin (1), hypophyllanthin (2), niranthin (3), and corilagin (4). To gain insights into the binding interactions of the most active compound hypophyllanthin (2), molecular docking was conducted and found that hypophyllanthin (2) exhibited favorable binding in the H1R binding site. In conclusion, P. amarus and hypophyllanthin (2) could potentially exhibit antiallergic activity by preventing the activation of the H1 receptor.

Effects of Phyllanthus amarus extract on nonspecific immune responses, growth, and resistance to Vibrio alginolyticus in white shrimp Litopenaeus vannamei

This study investigates the effects of Phyllanthus amarus extract (PAE) on immune responses, growth, and resistance to Vibrio alginolyticus in white shrimp (Litopenaeus vannamei). In vitro PAE treatment did not alter the cell viability of haemocytes and significantly enhanced immune parameters such as phenoloxidase (PO) activity, phagocytic activity, and superoxide anion (O2−) production. We conducted two feeding trials to examine the effects of PAE on the growth, disease resistance, and innate immune parameters of white shrimp. In the first in vivo trial, shrimps (4.01 ± 0.03 g) were fed a diet containing 0 g (control), 10 g (PAE10), 20 g (PAE20), or 40 g (PAE40) of PAE per kilogram of feed for 56 days. After the feeding period, the PAE20 group showed a significantly higher weight gain and specific growth rate than shrimp fed the control diet. Furthermore, after challenge with V. alginolyticus, shrimp fed a diet containing PAE showed significantly higher survival than those fed the control diet. The second in vivo trial (28 days) was performed to identify the mechanisms of enhanced immunity in PAE-fed shrimp. Shrimp fed the PAE20 diet generally had the highest total haemocyte count, PO activity, phagocytic activity, and O2− production, followed by the PAE40 and PAE10 groups. Thus, our results suggest that administration of 20 g of PAE per kilogram of feed can enhance immunity, growth, and resistance to V. alginolyticus in white shrimp.

Effect of &lt;i&gt;Phyllanthus amarus&lt;/i&gt; Coating Denture Resin on Candida Adhesion and its Effect on Human Gingival Fibroblast

Candida infection has become increasingly important as an opportunistic infection frequently found in the oral cavity of denture wearing patients. The infection develops on oral mucous membrane and can spread to other parts of the body. Phyllanthusamarus, extensively grown in many tropical and subtropical countries, has been reported to have various medicinal properties including antimicrobial activity. This study aimed to assess the effect of P.amarus coating denture resin on Candida adhesion and its effect on human gingival fibroblast. The P. amarus coating solution was prepared by water extraction of dried leaves and freeze drying. Denture PMMA resin (SCG Chemical, Thailand) samples were made and coated with various concentrations (0.01-10 mg/mL) of P. amarus solution. Candida adhesion assay was performed on C. albicans ATCC 10231, C. glabrata ATCC 6258, C. krusei ATCC 90030 and C. tropicalis ATCC 13803 based on the method described by Samaranayake and MacFarlane. Cytotoxicity test was done on human gingival fibroblasts (HGF) according to ISO standard 10993-5/2009 for medical product testing. The result showed that P.amarus coating denture resin exhibited an inhibitory effect on the adhesion of all species of test Candida. Cytotoxicity testing revealed a dose-dependent effect of the P. amarus extract on human gingival fibroblast. At the concentrations of 0.1, 1, 5 and 10 mg/mL, the extract produced 84.7%, 73.4%, 64.7% and 38.2% of cell viability, respectively. In conclusion, coating denture resin with P. amarus at a concentration of 1 mg/mL can be used safely as a natural alternative preventive to control or prevent Candida infection caused by C. krusei and C. tropicalis in denture-wearing patients avoiding side effects of chemical antifungal agents. Further studies are needed to define the antifungal components and mechanisms of action as well as clinical trials in the patients.

Lignans and Polyphenols of Phyllanthus amarus Schumach and Thonn Induce Apoptosis in HCT116 Human Colon Cancer Cells through Caspases-Dependent Pathway.

The anticancer effects of Phyllanthus amarus extract on various cancer cells have been investigated, however, the effects of its major constituents on HCT116 human colorectal cancer cells have not been reported. In the present study, we investigated the cytotoxic effect of 80% ethanol extract of P. amarus and its marker constituents (phyllanthin, hypophyllanthin, gallic acid, niranthin, greraniin, phyltetralin, isolintetralin, corilagin and ellagic acid) on HCT116 and their underlying mechanisms of action. Their antiproliferative and apoptotic effects on HCT 116 were performed using MTT assay and flow cytometric analysis, respectively, while caspases 3/7, 8 and 9 activities were examined using the colorimetric method. The expression of cleaved poly ADP ribose polymerase enzyme (PARP) and cytochrome c proteins was investigated by the immune-blot technique. HPLC and LC-MS/MS analyses demonstrated that the extract contained mainly lignans and polyphenols. The plant samples markedly suppressed the growth and expansion of HCT116 cells in a concentration- and time-dependent manner with no toxicity against normal human fibroblast CCD18 Co. P. amarus extract, phyllanthin and gallic acid induced mode of cell death primarily through apoptosis as confirmed by the exteriorization of phosphatidylserine. Caspases 3/7, 8, and 9 activities increased in a concentration-dependent manner following 24h treatment. The expressions of cleaved PARP (Asp 214) and cytochrome c were markedly upregulated. P. amarus extract, phyllanthin and gallic acid exhibited an apoptotic effect on HCT116 cells through the caspases-dependent pathway.