Secondary Metabolites of Endophytic Fungus Trichoderma harzianum Isolated from Physalis angulata / 中国实验方剂学杂志

Objective:To study the secondary metabolites of the endophytic fungus Trichoderma harzianum,which was isolated from Physalis angulata. Method:The strains were cultured in a big-scale by oscillating incubator. Then the obtained culture liquid and mycelia were extracted by EtOAC and actone,respectively. The extracts were integrated after recovery of solvents, and then the active secondary metabolites were isolated and purified by comprehensive use of open ODS flash column,sephadex LH-20,HPLC,and LC-MS analysis techniques. Their structures were identified according to their physico-chemical properties and nuclear magnetic resonance (NMR) data. The cytotoxicities of the compounds were determined by methyl thiazolyl tetrazolium(MTT) assay. Result:Ten compounds isolated from T. harzianum were identified as destruxin A2(1),destruxin B2(2),3-isobutyl-pyrrolopiperazine-2,5-dione[cyclo (leu-pro dipeptide)](3),cyclo (Phen-pro) dipeptide(4),cyclonerodiol(5),brevianamide F(6),N-acetyltryptamine(7),9-hydroxyl-(2-methylpropyl) isobutyl phthalate(8),5-hydroxy-3-hydroxymethyl-2-methyl-7-methoxychromone(9),and phenylpropionic acid(10). Compounds 1-10 didn't exhibit significant cytotoxicity to lung cancer cell line A549 in the MTT assay (IC50 ≥ 20 mg·L-1). Conclusion:All compounds were isolated from T. harzianum for the first time.

The "target + activity" method for rapid discovery of active compounds targeting Hsp90 in pancreatic cancer cells from Physalis angulata L / 药学学报

The purpose of this study was to select the active compounds targeting Hsp90 protein in pancreatic cancer cells through a new dual "target + activity" rapid discovery technique. We combined an in vitro anti-cancer activity screening method with a dual-luciferase reporter gene and multi-chromatography separation technology, for rapid discovery of potential Hsp90 inhibitors from the Chinese herbal medicine Physalis angulata L. The anti-proliferation activity of those compounds was assessed in pancreatic cancer cell line BxPC-3 by MTT assays. The molecular mechanisms of Hsp90 inhibition were explored by Western blot and shRNA knockdown assays. As a result, two withanolides, withanolide E (WE) and 4β-hydroxywithanolide E (HWE), were identified from Physalis angulata L. The half maximal inhibitory concentration (IC50) of WE and HWE were 0.71±0.03 and 1.23±0.10 μmol·L-1 for the growth of BxPC-3 cells in 48 h. Luciferase reporter assay demonstrated that WE and HWE significantly induced heat shock element (HSE) activity in a dose- and time-dependent manner. The molecular mechanism study showed that after exposing to 5 μmol·L-1 WE or HWE for 48 h, the aggregation of Hsp90 dimer was upregulated to 6.5±1.3 and 11.8±2.0 fold, while the expression of Hsp90 client protein Akt was downregulated to 21.7%±2.8% and 9.8%±1.4% of the control group. Moreover, the Hsp90 inhibitory activity of WE or HWE was canceled by shRNA mediated Hsp90 knockdown. Overall, based on the dual "target + active" rapid discovery technique, two new Hsp90 inhibitors WE and HWE were found from Physalis angulata L. The Hsp90 inhibitory mechanism of WE and HWE may be mediated by induction of Hsp90 aggregate dimer and inhibition of Hsp90 client protein Akt expression.

In vitro antileishmanial effects of Physalis angulata root extract on Leishmania infantum / 中西医结合学报

<p><b>OBJECTIVE</b>In the present study, we evaluated the effects of the aqueous extract of Physalis angulata root (AEPa) on Leishmania infantum proliferation, morphology, and the driving mechanism in leishmanicidal activity and modulatory action on macrophages.</p><p><b>METHODS</b>L. infantum promastigotes were treated with 50 and 100 µg/mL AEPa for 72 h and then antipromastigote assay was performed by counts in a Newbauer chamber, morphological changes were analyzed by transmission electron microscopy and the mechanism of the leishmanicidal activity was detected. In addition, macrophages were infected with L. infantum and were used to evaluate anti-amastigote activity of AEPa and effects of AEPa on cytokine secretion after 72-hour treatment.</p><p><b>RESULTS</b>Treatment with AEPa reduced the numbers of L. infantum promastigotes (50% inhibitory concentration (IC) = 65.9 μg/mL; selectivity index (SI) = 22.1) and amastigotes (IC = 37.9 μg/mL; SI = 38.5) compared with the untreated control. Amphotericin B reduced 100% of the promastigote numbers after 72 h of treatment (IC = 0.2 μg/mL). AEPa induced several morphological changes and increased the production of reactive oxygen species and apoptotic death in promastigotes after treating for 72 h. AEPa (100 μg/mL) promoted tumor necrosis factor-α secretion in macrophages infected with L. infantum after 72 h of treatment, but did not induce an increase in this cytokine in noninfected macrophages. In addition, AEPa showed no cytotoxic effect on J774-A1 cells (50% cytotoxic concentration >1000 μg/mL).</p><p><b>CONCLUSION</b>AEPa presented antileishmanial activity against the promastigotes and amastigotes of L. infantum without macrophage cytotoxicity; these results show that natural products such as P. angulata have leishmanicidal potential and in the future may be an alternative treatment for leishmaniasis.</p>

Chemical constituents from whole herb of Physalis angulata and their cytotoxic activity / 中草药

Objective: To investigate the chemical constituents from whole herb of Physalis angulata and test their cytotoxic activity. Methods: Compounds were isolated by chromatography methods and LC-MS guided analysis, structures were determined by spectroscopic techniques of 1D- and 2D-NMR (HSQC, HMBC, 1H-1H COSY, and NOESY), cytotoxic activity experiments were conducted by MTT method. Results: Twelve compounds were identified as physalins A (1), B (2), C (3), D (4), F (5), H (6), I (7), O (9), 5α-ethoxy-6β-hydroxy-5,6-dihydrophysalin B (8), β-sitosterol (10), 2-carboxyoxanilic acid methyl ester (11), and acetaminobenzoic acid (12). Conclusion: Among these compounds, compound 11 is a new natural product. Compounds 1-3 and 9 are firstly obtained from P. angulata. All compounds are tested for their inhibitory activities against human lung adenocarcinoma strain A-549, with IC50 values of 1.9-20.2 μmol/L, which indicates good cytotoxic activity of these physalins.

Influência da adubação fosfatada sobre o crescimento do camapu (Physalis angulata L. ) / Influence of phosphorus fertilization over the growth of the camapu (Physalis angulata L. )

RESUMO A espécie Physalis angulata L., pertencente à família Solanaceae, tem despontado como uma planta extremamente promissora para uso medicinal, em razão da produção do composto fisalina. No entanto, aspectos importantes do seu cultivo ainda permanecem pouco conhecidos. Assim, objetivou-se avaliar a influencia do fósforo para o seu crescimento. O experimento foi conduzido em Latossolo Amarelo de textura média, submetido a três doses de P: 8, 16 e 64 mg kg-1 de P. Usou-se um delineamento experimental inteiramente casualizado. Foram realizadas duas colheitas: aos 30 e 42 dias após o plantio. Verificou-se que as plantas cultivadas sob deficiência (8 mg kg-1 de P no solo) apresentaram 26,3 folhas, enquanto as cultivadas com a maior dose (64 mg kg-1 de P no solo) produziram 80,17 folhas. Aumento semelhante (201%) foi obtido para a área foliar, cujos valores variaram de 436,4 cm2a 1.313,9 cm2. Sob deficiência, a produção da massa seca total foi de apenas 1,72g, enquanto para a dose de 64 mg kg-1 de P no solo esse valor foi de 7,58g, significando um aumento de 340%. Plantas cultivadas com 8 mg kg-1 de P no solo não apresentaram flores ou frutos; já para o tratamento em que se utilizou 64 mg kg-1 de P no solo, o início do florescimento antecedeu ao das plantas cultivadas com 16 mg kg-1 de P no solo. A razão de área foliar e a área foliar específica foram maiores para as plantas cultivadas sob deficiência, enquanto a taxa de crescimento relativo e a taxa assimilatória líquida não foram estatisticamente influenciadas. Conclui-se que a deficiência de P no solo pode comprometer sobremaneira o crescimento das plantas de Physalis. Além disso, como a deficiência de P reduziu sensivelmente a produção de folhas, órgão responsável pela produção da fisalina, também é possível concluir que a deficiência de P pode, mesmo que indiretamente, reduzir a concentração desse importante composto na planta.

ABSTRACT The specie Physalis angulata L., who belongs to the Solanaceae family, has stand out as an extremely promising plant in terms of medical use, due to the production of the physalin compound. However, important aspects of its production still remain unknown. Thus the objective of this work was to assess the influence of phosphorus for the growth of these species. The experiment was performed with three doses of P (8, 16 e 64 mg kg-1 de P), in a completely randomized experimental design. Two harvests were performed: at 30 and 42 days after planting. The deficiency of P reduced the number of leaves and leaf area. The total dry matter (DM), as well as a MS from all parts of the plant, was significantly higher for plants grown under the highest levels of phosphorus. Plants grown with 8 mg P kg-1 showed no flowers or fruit; but, for the treatment which used 64 mg of P kg-1 the beginning of flowering happened before than with the plants grown with 16mg. The ratio of the leaf area and the specific leaf area were higher for plants grown under stress, while the values of the relative growth rate and the net assimilatory rate were not statistically influenced. We conclude that the deficiency of P in soil may considerably affect the growth of Physalis plants. Moreover, since the deficiency of P significantly reduced the leaf production, which is responsible for creating the physalin, it is also possible to conclude that P deficiency may, even indirectly, reduce the concentration of this important compound in the plant.

Physalis angulata L. (Bolsa Mullaca): a review of its traditional uses, chemistry and pharmacology / Physalis angulata L. (Bolsa Mullaca): revisión de usos tradicionales, química y farmacología

Physalis angulata is a specie of the Solanaceae family, which edible fruit is used in several countries of tropical and subtropical regions of the world as medicinal and fruit-tree. This review shows research over the last 30 years, about traditional uses, chemical constituents and pharmacology of this specie. The studies related to traditional uses show that P. angulata is known for its antimalarial, anti-inflammatory and post-partum treating properties. It presents the different pharmacological experiments in vitro and in vivo models that have been made, also the identification of phytochemical constituents with medicinal importance, the main being physalins and withanolides. Pharmacological studies have shown antiparasitic, anti-inflammatory, antimicrobial, antinociceptive, antimalarial, antileishmanial, immunosuppressive, antiasthmatic diuretic, and antitumor activities, thus validating its traditional uses and demonstrating the great potential of this specie for further development within the pharmaceutical industry.

Physalis angulata, es una especie de la familia Solanaceae, de frutos comestibles, que en diferentes países de regiones tropicales y subtropicales del mundo utilizan como medicinal y frutal. La presente revisión muestra las investigaciones realizadas durante los últimos 30 años, sobre los usos tradicionales, componentes químicos y farmacología de esta especie. Los estudios referidos a los usos tradicionales, muestran que la especie es conocida por propiedades antimaláricas, antiinflamatorias y en el tratamiento de postparto. Se muestran los diferentes experimentos farmacológicos de ensayos in vitro y modelos in vivo que se han realizado, asimismo la identificación de sus constituyentes fitoquímicos con importancia medicinal, siendo los principales las fisalinas y los witanólidos. Los estudios farmacológicos revelan que tiene actividad antiparasitaria, antiinflamatoria, antimicrobiana, antinociceptiva, antimalárica, antileishmania, inmunosupresor, antiasmático, diurético y antitumoral, validando de esta manera sus usos tradicionales y demostrando el gran potencial que tiene esta especie para un mayor desarrollo dentro de la industria farmacéutica.

Chemical constituents of whole plant of Physalis angulata L / 中国药学杂志

OBJECTIVE: To study the chemical constituents of whole plant of Physalis angulata L. METHODS: The constituents were isolated and purified by chromatographic methods and their structures were elucidated by spectroscopic methods and physio-chemical analysis. RESULTS: Thirteen known compounds was isolated and identified as physalin A (1), physalin B (2), physalin E (3), physalin P(4), stigmast-5-en-3β-ol (5), ergost-5, 24 (28) -diene-3β-ol (6), brassicaaterol (7), stigmasta-22-en-3, 6-dione (8), pregn-5-en-3-ol-20-carboxylic acid (9), ergost-5, 24 (28)-diene-3β, 23S-diol (10), ergosta-5, 25 (26)-diene-3, 24ξ-diol(11), n-hexade-canoic acid (12), and n-heptadecanoic acid (13). CONCLUSION: All of the compounds except 2 and 3 are isolated from this plant for the first time, and compounds 5-10 and 12-13 are isolated from Physalis for the first time.

Atividades antimicrobiana e fototóxica de extratos de frutos e raízes de Physalis angulata L. / Antimicrobial and phototoxic activities of Physalis angulata L (Solanaceae) extracts, fruits and roots

Extratos e frações de frutos e raízes de Physalis angulata L. (Solanaceae) foram ensaiados para encontrar atividade antimicrobiana. Aplicando o método de difusão em agar, todas as amostras foram testadas contra Staphylococcus aureus ATCC 6538. O extrato etanólico dos frutos apresentou atividade antibacteriana, a qual teve a atividade fototóxica estimada em cobaias quando expostas a luz ultravioleta, e não foram observados eritemas. Esses dados impulsionaram a pesquisar diferentes formas de obtenção de extratos da planta, com o objetivo de preparar formulações com atividade anti-séptica, que possam se apresentar mais eficazes e seguras, quando aplicadas no tratamento de doenças infecciosas.

Extracts and fractions of Physalis angulata L. prepared from fruits and roots were assayed to find out antimicrobial activity. Using the agar diffusion method all samples were tested against Staphylococcus aureus ATCC 6538. The ethanolic extract of the fruits displayed bacterial activity. Phototoxic property was estimated with guinea pigs when they were exposed to ultraviolet light, no erythemas were observed. These data encouraged us to look for different forms of extracts wich could be applied as a safe and effective antiseptic product.

Estudo farmacobotânico comparativo entre Nicandra physalodes e Physalis angulata (Solanaceae) / Comparative pharmacobotanical study on Nicandra physalodes and Physalis angulata (Solanaceae)

Neste trabalho realizou-se uma caracterização morfoanatômica comparativa, entre as folhas de Nicandra physalodes (L.) Gaertn. e Physalis angulata L. (Solanaceae), espécies de uso na medicina popular, cujas folhas e raízes são indicadas como diurético e antitérmico. As identificações botânicas e as morfodiagnoses macroscópicas foram realizadas após estudos morfológicos. Para as morfodiagnoses microscópicas realizaram-se cortes paradérmicos e transversais das folhas (lâmina e pecíolo), clarificados e posteriormente corados com safranina e safrablue. Além dos caracteres morfológicos da flor e do fruto, as duas espécies diferem pelos seguintes caracteres: vascularização da nervura principal em forma de arco em N. physalodes, e semicircular em P. angulata; drusas no mesofilo de P. angulata; tricomas glandular-estipitados com a glândula apical bisseriada em N. physalodes. As duas espécies distinguem-se principalmente pela morfologia da flor e fruto e pela anatomia do mesofilo e vascularização da nervura principal.

In this work a morphoanatomical comparative characterization was carried out between the leaves of Nicandra physalodes (L.) Gaertn. and Physalis angulata L. (Solanaceae) species used in folk medicine, whose leaves are indicated as diuretic and febrifuge. The botanical identification and macroscopical morphodiagnosis were made after morphological studies. The microscopical morphodiagnosis were carried out by paradermic and transversal cuts of the leaves (blade and petiole), clarified and coloured with safranin and astrablue. Moreover the morphological characters of the flower and fruit, of the two species can be distinguished by the following characters: N. physalodes has the bundle vascular of the middle vein in arc format; P. angulata has the vascular bundle of the middle vein in a semi-circle format; druses in the mesophyll of P. angulata; andthe presence of trichomes glandular-estipitate with bisseriate head in N. physalodes. The morphology of flower and fruit and the anatomy of mesophyll, the vascular bundles of middle vein are the principal distinctive characters between them.