Inhibitory Effects of the Standardized Extract of Phyllanthus amarus on Cellular and Humoral Immune Responses in Balb/C Mice

Phyllanthus amarus has been shown to have strong inhibitory effects on phagocytic activity of human neutrophils and on cellular immune responses in Wistar–Kyoto rats. In this study, we investigated the effects of daily treatment of standardized extract of P. amarus at 50, 100 and 200 mg/kg for 14 days in Balb/C mice by measuring the myeloperoxidase activity (MPO), nitric oxide (NO) release, macrophage phagocytosis, swelling of footpad in delayed type hypersensitivity (DTH), and serum immunoglobulins, ceruloplasmin and lysozyme levels. Qualitative and quantitative analyses of the extract using validated reversed‐phase HPLC methods identified phyllanthin, hypophyllanthin, corilagin and geraniin as the biomarkers. Significant dose‐dependent inhibitions of MPO activity and NO release were observed in treated mice. The extract also inhibited E. coli phagocytic capacity of peritoneal macrophages of treated mice and inhibited the sheep red blood cells (sRBC)‐induced swelling rate of mice paw in the DTH. There was also a significant decrease in non‐specific humoral immunity including ceruloplasmin and lysozyme levels in the extract‐fed groups as well as the release of serum level immunoglobulins. The strong inhibitory effects of the extract on the cellular and humoral immune responses suggest the potential of the plant to be developed as an effective immunosuppressive agent. Copyright © 2016 John Wiley & Sons, Ltd.     

Comparative pharmacognostic studies of three Phyllanthus species

Different species of Phyllanthus are considered to be very effective hepatoprotective agents in the Indian indigenous systems of medicine and are considered bitter, astringent, stomachic, diuretic, febrifuge, deobstruant and antiseptic. Still ayurvedic practitioners prescribed fresh juice of 'Bhuiamlki' for jaundice. Various species of Phyllanthus are being sold in India under the trade name 'Bhuiamlki'. During market surveillance of herbal drug, it was observed that almost all the commercial samples, either comprise of Phyllanthus amarus Schum & Thonn. or Phyllanthus maderaspatensis Linn. or mixture of Phyllanthus amarus, Phyllanthus fraternus Webster. and Phyllanthus maderaspatensis. Therefore, in this context the detailed pharmacognostical evaluation of all the three species has been carried out with the aim to establish the identification markers of this important hepatoprotective agent (effective in hepatitis B too). The study conclude that all the three species can be differentiated on the basis of macro and microscopic characters, physico-chemical values, HPTLC fingerprint profile, and detection of phyllanthin and hypophyllanthin as marker components. Besides, an interesting conclusion can also be drawn that phyllanthin and hypophyllanthin said to protect hepatocytes against carbon tetrachloride and galactosamine induced toxicity, may not be exclusively responsible for hepatoprotective activity as these are present only in Phyllanthus amarus while Phyllanthus fraternus and Phyllanthus maderaspatensis also possess significant hepatoprotective activity.     

固定化介导提高苦味叶下珠中的海冰叶下珠脂素和叶下珠次素的含量(英文)

Phyllanthus amarus plant is used in the traditional system of medicine as a hepatoprotective drug for which the majorlignans phyllanthin and hypophyllanthin are responsible.So far,no significant work has been done on the culture of this plant.Realizingthe hepatoprotective potential,the present investigation was undertaken.A cost effective process was developed for enhancingphyllanthin and hypophyllanthin utilizing the immobilization technique.HPTLC was used to compare the phyllanthin and hypophyllanthincontents in calcium alginate immobilized cells obtained from fresh grown plants and MS medium was supplemented withdifferent abiotic elicitors,under aseptic conditions for the treatment with chitosan,copper sulphate,phenylalanine and silver nitratesolution to make the whole process commercially viable.It was revealed that silver nitrate and phenylalanine at low concentrationenhances phyllanthin and hypophyllanthin yield as compared to control immobilized cell culture.The study revealed that an increase inthe content of phyllanthin and hypophyllanthin was elicitor concentration dependent and silver nitrate treatment gave a maximum yieldof hepatoprotective bioactives as compared to the other abiotic elicitors used.     

印度、海南、河南产叶下珠属治疗乙型肝炎的疗效观察

采用河南省蜜柑草PhyllanthusurinariaL.海南省小返魂P.niruriL.及印度提供的苦味叶下珠P.amarusL.治疗乙肝患者各35例、42例、11例。结果P.urinariaL.组对HBeAg有转阴作用,对HBeAb有转阳作用,其余2种皆无这种作用。上述3种对HBsAg转阴皆无作用。     

Mechanisms of antihyperuricemic effect of Phyllanthus niruri and its lignan constituents

Ethnopharmacological relevance

Phyllanthus niruri Linn. (Euphorbiaceae) is used as folk medicine in South America to treat excess uric acid. Our initial study showed that the methanol extract of Phyllanthus niruri and its lignans were able to reverse the plasma uric acid of hyperuricemic animals.

Aim of the study

The study was undertaken to investigate the mechanisms of antihyperuricemic effect of Phyllanthus niruri and its lignan constituents.

Material and methods

The mechanisms were investigated using xanthine oxidase assay and uricosuric studies in potassium oxonate- and uric acid-induced hyperuricemic rats.

Results

Phyllanthus niruri methanol extract exhibited in vitro xanthine oxidase inhibition with an IC₅₀ of 39.39 μg/mL and a moderate in vivo xanthine oxidase inhibitory activity. However, the lignans display poor xanthine oxidase inhibition in vitro and a relatively weak in vivo inhibitory activity at 10 mg/kg. On the other hand, intraperitoneal treatment with Phyllanthus niruri methanol extract showed 1.69 folds increase in urinary uric acid excretion when compared to the hyperuricemic control animals. Likewise, the lignans, phyllanthin, hypophyllanthin and phyltetralin exhibited up to 2.51 and 11.0 folds higher in urinary uric acid excretion and clearance, respectively. The co-administration of pyrazinamide with phyllanthin exhibited a significant suppression of phyllanthin's uricosuric activity resembling that of pyrazinamide with benzbromarone.

Conclusions

The present study showed that the antihyperuricemic effect of Phyllanthus niruri methanol extract may be mainly due to its uricosuric action and partly through xanthine oxidase inhibition, whereas the antihyperuricemic effect of the lignans was attributed to their uricosuric action.     

余甘子及其活性成分肝保护作用及机制的研究进展

余甘子来源于余甘子Phyllanthus emblica的干燥成熟果实,是我国重要的药食同源品种,具有抗炎、抗氧化、抗肿瘤和免疫调节等广泛的生物活性.通过对余甘子提取物及其所含没食子酸、鞣花酸及柯里拉京等活性成分的药理作用进行总结,发现其对肝损伤、病毒性肝炎、非酒精性脂肪肝、肝纤维化及肝癌等具有治疗作用,主要通过抑制脂...     

Antihyperalgesic and Antiedematous Activities of Bisabolol‐Oxides‐Rich Matricaria Oil in a Rat Model of Inflammation

From the dried flower heads of Matricaria recutita L., essential oil was isolated by hydrodistillation, and in the obtained blue oil, α‐bisabolol oxide A (21.5%), α‐bisabolol oxide B (25.5%) and (Z)‐spiroether (cis‐en‐yn‐spiroether) (10.3%) were identified as the main compounds, by gas chromatography (GC) and GC–mass spectrometry analyses. The antihyperalgesic effects of this oil were examined in a rat model of inflammation induced by carrageenan, through a modified ‘paw‐pressure’ test. Antiedematous effects were examined in a rat model of inflammation induced by carrageenan, dextran and histamine, through plethysmometry. Matricaria oil (25, 50 and 100 mg/kg, p.o.) exhibited a significant dose‐dependent reduction of hyperalgesia and edema induced by carrageenan in both prophylactic and therapeutic treatment schemes. It was more efficacious in the prophylactic treatment scheme, and the corresponding median effective dose (ED₅₀) ± standard error of the mean (SEM) values were 49.8 ± 6.0 and 42.4 ± 0.2 mg/kg for antihyperalgesic and antiedematous effects, respectively. Prophylactic treatments with matricaria oil (25, 50 and 100 mg/kg, p.o.) caused a significant dose‐dependent antiedematous effect in dextran‐induced edema with lower efficacy than in the carrageenan model. In a dose of 100 mg/kg, p.o., matricaria oil caused a slight reduction of histamine‐induced edema. These results suggest that bisabolol‐oxide‐rich matricaria oil may be effective against pain and edema present in various inflammatory conditions, which supports matricaria traditional uses. Copyright © 2013 John Wiley & Sons, Ltd.     

Protective activity of phyllanthin in ethanol-treated primary culture of rat hepatocytes

Aim of the study

To investigate the protective effect of phyllanthin (a known principal constituent of Phyllanthus amarus Schum. et Thonn.) on ethanol-induced rat liver cell injury.

Materials and methods

Primary culture of rat hepatocytes (24 h culturing) were pretreated with phyllanthin (1, 2, 3 and 4 μg/ml) for 24 h. After 24 h pretreatment, cells were treated with ethanol (80 μl/ml) for 2 h.

Results

Ethanol decreased %MTT, increased the release of transaminases (ALT and AST) with the increase in the production of intracellular ROS and lipid peroxidation. Phyllanthin demonstrated its role in protection by antagonizing the above effect induced by ethanol. Phyllanthin also restored the antioxidant capability of rat hepatocytes including level of total glutathione, and activities of superoxide dismutase (SOD) and glutathione reductase (GR) which were reduced by ethanol.

Conclusions

These results suggested the hepatoprotective effect of phyllanthin against ethanol-induced oxidative stress causing rat liver cell damage through its antioxidant activity.     

Involvement of Cerebral Nervous System Areas and Cytokines on Antihyperalgesic and Anti‐Inflammatory Activities of Kielmeyera rugosa Choisy (Calophyllaceae) in Rodents

Kielmeyera rugosa is a medicinal plant known in Northeastern Brazil as ‘pau‐santo’, and it is used in the treatment of several tropical diseases such as malaria, schistosomiasis, and leishmaniasis. We evaluated antihyperalgesic and anti‐inflammatory activities of methanol stem extract of K. rugosa (MEKR) in mice. The mechanical hyperalgesia induced by carrageenan and tumor necrosis factor‐alpha (TNF‐α), prostaglandin E₂, and dopamine were assessed. We also investigated the anti‐inflammatory effect of MEKR on carrageenan‐induced pleurisy and paw edema. Ninety minutes after the treatment, the animals were submitted to an imunofluorescence for Fos protein. MEKR (100, 200, and 400 mg/kg; p.o.) inhibited the development of mechanical hypernociception and edema. MEKR significantly decreased TNF‐α and interleukin 1β levels in pleural lavage and suppressed the recruitment of leukocytes. MEKR (1, 10, and 100 mg/mL) did not produce cytotoxicity, determined using the methyl‐thiazolyl‐tetrazolium assay in vitro. The locomotor activity was not affected. MEKR activated significantly the bulb olfactory, piriform cortex, and periaqueductal gray of the central nervous system. Our results provide first time evidence to propose that MEKR attenuates mechanical hyperalgesia and inflammation, in part, through an activation of central nervous system areas, mainly the periaqueductal gray and piriform cortex areas. Copyright © 2014 John Wiley & Sons, Ltd.     

Evaluation of the acute toxicity,analgesic and CNS activities of different species of Teucrium genus

Methanol and dichloromethanol extracts of the leaves and stems of four Teucrium species (T. cartaginenses, T. flavum, T. pumillum and T. buxifolium) have been tested for their toxicity, analgesic and central depressor effects. The intraperitoneal administration of the different extracts showed a CNS depressant activity in mice, but they lacked anticonvulsive effects. When tested for analgesic activity none of the extracts increased the threshold of pain thermal stimulus. However, the methanol and dichloromethanol extracts of T. cartaginenses and T. buxifolium species showed a significant analgesic effect in models of pain induced by chemical or mechanical stimulation.     

Ethnopharmacology guided screening of traditional Indian herbs for selective inhibition of Plasmodium specific lactate dehydrogenase

Ethnopharmacological relevance

Medicinal plants traditionally used to treat malaria can provide quality leads towards identifying novel anti-malarial drugs. Here we combined this approach with target based drug discovery and explored Plasmodium specific lactate dehydrogenase (LDH) inhibitory activity of 8 Indian plants which are ethnically used to treat malaria.

Methods

LDH from Indian Plasmodium falciparum and Plasmodium vivax strains, were cloned and expressed in Escherichia coli, followed by purification of recombinant enzymes (rPfLDH and rPvLDH respectively). Extracts of 8 plants in different organic and aqueous solvents, were screened for their inhibitory activity on rPfLDH, rPvLDH and mammalian LDHs. Phyllanthus amarus aqueous extract was further tested for in vitro parasiticidal activity.

Results

Aqueous extract of Phyllanthus amarus Schum. and Thonn. and chloroform extract of Murraya koenigii (L.) Spreng. exhibited profound and exclusive inhibitory effect on Plasmodium falciparum LDH (IC₅₀=11.2 μg/ml±0.4) and Plasmodium vivax LDH (IC₅₀=6.0 μg/ml±0.6) respectively. Moreover, Phyllanthus amarus aqueous extract also demonstrated antiplasmodial activity in vitro, on Chloroquine sensitive and resistant strains of Plasmodium falciparum (IC₅₀=7.1 μg/ml±0.5 and 6.9 μg/ml±0.7 respectively).

Conclusion

Target specific screening of traditional herbs used in malaria treatment has proffered Phyllanthus amarus and Murraya koenigii extracts as hits which can optimistically provide novel antimalarial drugs.     

Study of the antioedema activity of some seaweed and sponge extracts from the mediterranean coast in mice

Chloroform and methanol extracts of ten marine species, seven seaweeds and three sponges, have been studied for possible, antioedema activities. The extracts were administered either topically or orally on TPA-induced mouse ear oedema and on carrageenan mouse paw oedema, respectively. The most interesting seaweed extracts were found to be from Corallina elongata, Galaxaura oblongata, Laurencia obtusa and Udotea petiolata, where both extracts of each species induced a large antioedema effect in both models employed. None of the sponges assayed demonstrated antiinflammatory effects on carrageenan mouse paw oedema, however, some extracts elicited an inhibition of the oedema developed by TPA.     

Phyllanthus amarus: ethnomedicinal uses, phytochemistry and pharmacology: a review

Ethnopharmacological relevance

Phyllanthus amarus Schum. &; Thonn. belongs to the family Euphorbiaceae is a small herb well known for its medicinal properties and widely used worldwide. P. amarus is an important plant of Indian Ayurvedic system of medicine which is used in the problems of stomach, genitourinary system, liver, kidney and spleen. It is bitter, astringent, stomachic, diuretic, febrifuge and antiseptic. The whole plant is used in gonorrhea, menorrhagia and other genital affections. It is useful in gastropathy, diarrhoea, dysentery, intermittent fevers, ophthalmopathy, scabies, ulcers and wounds.

Materials and methods

The present review covers a literature across from 1980 to 2011. Some information collected from traditional Ayurvedic texts and published literature on ethanomedicinal uses of Phyllanthus amarus in different countries worldwide.

Results

Phytochemical studies have shown the presence of many valuable compounds such as lignans, flavonoids, hydrolysable tannins (ellagitannins), polyphenols, triterpenes, sterols and alkaloids. The extracts and the compounds isolated from P. amarus show a wide spectrum of pharmacological activities including antiviral, antibacterial, antiplasmodial, anti-inflammatory, antimalarial, antimicrobial, anticancer, antidiabetic, hypolipidemic, antioxidant, hepatoprotective nephroprotective and diurectic properties.

Conclusion

The present review summarizes information concerning the morphology, ecology, ethnopharmacology, phytochemistry, biological activities, clinical applications and toxicological reports of P. amarus. This review aims at gathering the research work undertaken till date on this plant in order to provide sufficient baseline information for future works and commercial exploitation.     

Effects of Ginkgo Biloba Extract EGb‐761 on Neuropathic Pain in Mice: Involvement of Opioid System

Neuropathic pain is considered as one of the most difficult types of pain to manage with conventional analgesics. EGb‐761 is extracted from leaves of Ginkgo biloba and has analgesia and anti‐inflammatory properties. This study aimed to examine the effect of EGb‐761 on chronic constriction injury (CCI)‐induced neuropathic pain behaviors, including thermal hyperalgesia and mechanical allodynia, and to explore the possible mechanisms underlying this action. To this end, CCI mice were intraperitoneally injected with EGb‐761 (10, 20, 40, and 80 mg/kg), and thermal hyperalgesia, mechanical allodynia, cytokines, and mu‐opioid receptor expression were measured. Results showed that EGb‐761 attenuated thermal hyperalgesia and mechanical allodynia dose‐dependently and the best delivery time window was from day 7 to day 14 after CCI. Additionally, EGb‐761 treatment significantly decreased pro‐inflammatory cytokines and enhanced mu opioid receptor (MOR) expression in the sciatic nerve. Moreover, the opioid antagonist naloxone prevented the effect of EGb‐761 on thermal hyperalgesia and mechanical allodynia but did not influence the effect of EGb‐761 on inflammatory cytokines. In conclusion, this study suggests that the potential of EGb‐761 as a new analgesic for neuropathic pain treatment, and opioid system may be involved in the EGb‐761‐induced attenuation of thermal hyperalgesia and mechanical allodynia. Copyright © 2016 John Wiley & Sons, Ltd.     

Anti-hyperalgesic activity of corilagin,a tannin isolated from Phyllanthus niruri L. (Euphorbiaceae)

Ethnopharmacological relevance

Corilagin (β-1-O-galloyl-3,6-(R)-hexahydroxydiphenoyl-d-glucose) is a tannin isolated from Phyllanthus niruri (Euphorbiaceae). This plant is well known for their therapeutic purposes to treat several diseases associated with dolorous process and are used in several ethno-medicines in tropical and subtropical countries.

Aim of the study

This study was designed to evaluate the anti-hyperalgesic activity of corilagin using chemically and thermally based nociception models in mice.

Materials and methods

Corilagin was isolated from Phyllanthus niruri (Euphorbiaceae) by extraction and chromatographic procedures and the anti-hyperalgesic activity was evaluated by using writhing, formalin, capsaicin, glutamate and hot plate tests in mice.

Results

Corilagin presented activity in acetic acid model with the ID₅₀ calculated value of 6.46 (3.09–13.51) being about 20.6 fold more potent than acetylsalicylic acid. It also exhibited activity against the first phase of formalin test with ID₅₀ value of 18.38 (15.15–22.59) μmol/kg. In the capsaicin and glutamate models, corilagin demonstrated significant activity at the 3 mg/kg.

Conclusion

The experimental data demonstrated that corilagin exhibits anti-hyperalgesic activity that may be due to interaction with the glutamatergic system.     

不同产地和采集季节叶下珠中鞣料云实精含量的比较

目的：对全国10个地区及同一地区不同采收期的叶下珠进行了鞣料云实精的含量测定比较。方法：分光光度法测定，测定波长270nm。结果：海南和贵州的鞣料云实精含量最低为0．58％，西安的含量最高达1．78％。采收期以8，9月份含量最高，为1．24％。结论：不同产地的叶下珠中鞣料云实精的含量有较大差异，且同一产地不同的采收期的鞣料云实精含量相差也较大。     

Protection afforded by aqueous extracts of Phyllanthus species against cytotoxicity induced by lead and aluminium salts

Oral administration of aqueous extracts of Phyllanthus emblica L. fruit and P. niruri L. leaves to laboratory bred albino mice for a week, significantly reduced the cytotoxic action of lead nitrate and aluminium sulphate. The frequency of chromosomal breakages, gaps and rearrangements induced by three concentrations of these salts was decreased when compared to the control animals which had received the salts alone. The plant extracts were equally effective in modifying the clastogenic effects of both lead nitrate and aluminium sulphate.     

Antinociceptive properties of extracts of new species of plants of the genus Phyllanthus (Euphorbiaceae)

The hydroalcoholic extract (HE) of the four new species of Phyllanthus, given intraperitoneally, produced significant inhibition of acetic acid-induced abdominal constrictions, with mean ID(50) values of 0.3, 1.8, 7.4 and 26.5 mg/kg for Phyllanthus amarus, Phyllanthus orbiculatus, Phyllanthus fraternus and Phyllanthus stipulatus, respectively. In the formalin test, the four species of Phyllanthus, also produced graded inhibition against both phases of formalin-induced licking, being more active in relation of the late phase. The HE of the Phyllanthus species elicited significant inhibition of the capsaicin-induced neurogenic pain, with mean ID(50) values of 8.9, 6.7, >30 and approximately 30 mg/kg for P. amarus, P. fraternus, P. stipulatus and P. orbiculatus, respectively. Given orally all HE of the Phyllanthus species were less potent and efficacious than when given by intraperitoneally. Results of the present study extend previous data and indicate that all extracts of Phyllanthus plants so far studied exhibit pronounced antinociception when assessed in chemical models of nociception, namely acetic acid-induced writhing, and formalin and capsaicin-induced licking.     

不同产地叶下珠药材中总酚含量测定

目的:建立不同产地叶下珠药材中总酚的含量测定方法.方法:以柯里拉京为对照品,三氯化铁-铁氰化钾为显色剂,检测波长744 nm,采用UV测定11个不同产地叶下珠药材中总酚的含量.结果:柯里拉京在0.604 ～ 3.02 mg·L-1线性关系良好(r =0.999 1),平均加样回收率99.79％,RSD 2.72％.不同产地叶下珠药材中总酚质量分数5.00％ ～ 12.16％.结论:该方法简单准确、重复性好,可用于叶下珠药材中总酚含量的测定.     

针刀疗法对肩周炎兔血清和肌肉组织中5-羟色胺和前列腺素E_2含量的影响

目的观察针刀疗法对肩周炎兔血清和肌肉组织中5-羟色胺（5-HT）和前列腺素E2（PGE2）含量的影响,探讨针刀疗法对肩周炎干预作用的可能机制及介入时机。方法将40只雄性新西兰兔随机分为正常组、模型组、针刀A组（疼痛期干预）、针刀B组（僵硬期干预）、针刀C组（疼痛期及僵硬期均干预）,每组8只。应用持续劳损加冰敷方法制备肩周炎家兔模型,于实验第33日分别取血清、肱二头肌、冈上肌、冈下肌,检测5-HT和PGE2的含量。结果模型组血清和肌肉组织中5-HT和PGE2含量均较正常显著升高,差异有统计学意义（P〈0.01）;针刀各组较模型组均有不同程度的降低,针刀C组血清和肌肉组织中5-HT和PGE2含量较模型组均显著降低,差异有统计学意义（P〈0.05,P〈0.01）,优于针刀A组和针刀B组。结论针刀疗法对肩周炎模型兔血清和肌肉组织中的5-HT和PGE2具有一定的调节作用,疼痛期和僵硬期同时干预效果更佳。