Antifungal activity of Euphorbia species against moulds responsible of cereal ear rots.

AIMS: This work aimed to identify secondary metabolites from aerial parts of Euphorbia species functional for control of toxigenic Fusarium species responsible of cereal grain rots. METHODS AND RESULTS: Aerial parts of Euphorbia serpens, Euphorbia schickendantzii and Euphorbia collina were sequentially extracted with hexane, ethyl acetate and methanol. The extracts were tested against strains of Fusarium verticillioides and Fusarium graminearum by microdilution tests. The hexane extract of E. collina provided the lowest IC50 s on both fungal species. Further fractionation showed that cycloartenol (CA) and 24-methylenecycloartanol are associated to the moderate inhibitory effect of the hexane extract on fungal growth.Sublethal concentrations of CA and 24MCA blocked deoxynivalenol (DON) and fumonisins production.CA and 24MCA co-applied with potassium sorbate, a food preservative used for Fusarium control, synergized the growth inhibition of fungi. The mixtures reduced mycotoxins accumulation when applied at sublethal concentrations. CONCLUSIONS: CA and 24MCA inhibited both fungal growth and mycotoxins production. This fact is an advantage respect to potassium sorbate which increased the mycotoxins accumulation at sublethal concentrations. SIGNIFICANCE AND IMPACT OF THE STUDY: CA and 24MCA synergized potassium sorbate and their mixtures offer a lower mycotoxigenic risk than potassium sorbate for control of the Fusarium species.

Macrocyclic diterpenoids from the seeds of Euphorbia peplus with potential activity in inducing lysosomal biogenesis.

The first phytochemical investigation of the seeds of Euphorbia peplus led to the isolation and characterization of five new (1-5), named euphopepluanones A-E, and five known diterpenoids (6-10). Their structures were established by extensive spectroscopic analysis and X-ray crystallographic experiments. Euphopepluanones A-E (1-3) feature a very rare 5/11/5-tricyclic skeleton, and euphopepluanones D-E (4-5) represent the first report of lathyrane type diterpenoids found in E. peplus. The new compounds 1-5 were assessed for their activities to induce lysosomal biogenesis through LysoTracker Red staining, in which compounds 1 and 3 could significantly induce lysosomal biogenesis. In addition, compounds 1 and 3 could promote the nuclear translocation of TFEB, a master transcriptional factor of lysosomal genes, indicating that compounds 1 and 3 induced lysosomal biogenesis through activation of TFEB.

Cytotoxicity and Chromatographic Fingerprinting of Euphorbia Species Used in Traditional Medicine.

BACKGROUND AND OBJECTIVE: Chromatographic fingerprinting of plant species play an important role in species identification and standardization of plant based health products. Some of the Euphorbia species are used in folk medicine, yet majority of these exhibit various degrees of toxicity. It becomes a challenge to distinguish the toxic from the non-toxic species. The study aimed to evaluate cytotoxicity and to determine the method for fingerprinting the chemical constituents of the selected Euphorbia species to identify markers of toxicity. MATERIALS AND METHODS: Hexane, DCM, ethyl acetate and methanol extracts of E. arabica, E. bupleurifolia, E. enopla, E. gorgonis, E. horrida indigenous and E. horrida var. were examined in mammalian vero cell line using MTT cell viability test assay. The presence of secondary metabolites and proteins were assessed in the plant extracts and thin layer chromatography was used to identify toxicity markers. RESULTS: The hexane and DCM extracts of E. arabica, E. bupleurifolia and the DCM extract of E. horrida var. exhibited the highest cell growth inhibition reaching IC50 at a concentration of 10 µg mL-1. Both polar and non-polar extracts of E. enopla exhibited cell growth inhibition with the hexane extract reaching IC50 at a concentration of 10 µg mL-1. Euphorbia gorgonis and E. horrida indigenous were not active against the vero cell line. Secondary metabolites were detected, however, proteins were not detected in all six Euphorbia species. The TLC profiles of toxic extracts revealed additional bands which were absent in non-toxic species. CONCLUSION: It is concluded that the TLC method developed in this study can be used as a quick screen method to possibly distinguish toxic from non-toxic species, as well as in identifying the studied species.

Applications of Chromatographic Techniques for Fingerprinting of Toxic and Non-toxic Euphorbia Species.

BACKGROUND AND OBJECTIVE: Euphorbia species have historically been used as medicinal plants to treat different ailments. However, some species have been reported to exhibit various degrees of toxicity. It becomes critical to distinguish toxic species from those that are non-toxic, for a particular application. The aim of the study was to determine the method for fingerprinting the chemical constituents of the selected toxic and non-toxic Euphorbia species to identify markers of toxicity. MATERIAL AND METHODS: Hexane, DCM, methanol, ethyl acetate and water plant extracts of Euphorbia ammak, clavarioides, caerulescens, polygona and trigona were investigated for their cytotoxic activities towards the mammalian Vero cell line using MTT cell viability test assay. The presence of secondary metabolites and proteins were assessed in the plant extracts. Moreover, the study used chromatographic methods to fingerprint the plant extracts to identify toxicity markers. RESULTS: The DCM extract of E. ammak exhibited the highest cell growth inhibition at all concentrations tested. The non-polar extracts of E. clavarioides exhibited the highest cell growth inhibition activity with hexane extract reaching IC50 at 1 µg mL-1. The DCM extract of E. caerulescens reached IC50 at a concentration of 10 µg mL-1, while other extracts didn't show any activity. The hexane and DCM extracts of E. polygona exhibited the highest cell growth inhibition activity, reaching IC50 at a concentration of 10 µg mL-1. All 4 extracts of E. trigona didn't show cell growth inhibition. All Euphorbia species showed the presence of secondary metabolites. The biuret and xanthoprotein methods indicated that there were no proteins detected in all 5 Euphorbia species. TLC profiles of toxic extracts revealed additional bands which were absent in non-toxic species. CONCLUSION: It is concluded that the TLC method developed in this study can be used as a quick screen method to possibly distinguish toxic from non-toxic species, as well as in identifying the studied species.

Tumor Cell Death via Apoptosis and Improvement of Activated Lymphocyte Cytokine Secretion by Extracts from Euphorbia Hebecarpa and Euphorbia Petiolata.

Background: Immunomodulatory materials from natural herbs and the characterization of their immune enhancement effects may have tremendous potential as cancer treatment. The aim of the present study was to investigate the apoptosis-inducing activities of Euphorbia hebecarpa Boiss and Euphorbia petiolata Banks & Sol. plant extracts and their effects on cytokine secretion by lymphocytes. Materials and Methods: We assessed the apoptosis-inducing effect of the plants' hexane extracts on previously determined sensitive cell lines (HeLa for E. hebecarpa and K562 for E. petiolata) by flow cytometry and measurement of caspase 3 activation. The apoptosis-related gene expressions were examined by real-time PCR. The effects of the extracts on lymphocyte proliferation and cytokine secretion were examined. Results: Flow cytometry analysis showed that the inhibitory effect of the extracts on tumor cell growth was due to cell apoptosis. The plant extracts at the 100 µg/ml dose induced apoptosis in HeLa (98.5 ± 0.1%) and K562 (57.7 ± 1.9%) cells. The extracts increased caspase 3 activation (≈2-fold>control). Real-time PCR showed Fas and Bax gene upregulation and Bcl-2 downregulation, which resulted in an increased Bax/Bcl-2 expression ratio. The extracts increased lymphocyte proliferation and increased levels of IFN-γ production in the presence and absence of mitogen (p < 0.05). They significantly increased IL-4 and decreased IL-10 secretion by mitogen-stimulated lymphocytes. E. hebecarpa also increased IL-17 release. Conclusion: These results have shown that both extracts possess antitumor activity by inducing apoptosis, possibly through both intrinsic and extrinsic pathways. In addition, they induced secretion of different T helper subset related cytokines that are effective in the immune response against cancer.

Evolutionary prediction of medicinal properties in the genus Euphorbia L.

The current decrease of new drugs brought to the market has fostered renewed interest in plant-based drug discovery. Given the alarming rate of biodiversity loss, systematic methodologies in finding new plant-derived drugs are urgently needed. Medicinal uses of plants were proposed as proxy for bioactivity, and phylogenetic patterns in medicinal plant uses have suggested that phylogeny can be used as predictive tool. However, the common practice of grouping medicinal plant uses into standardised categories may restrict the relevance of phylogenetic predictions. Standardised categories are mostly associated to systems of the human body and only poorly reflect biological responses to the treatment. Here we show that medicinal plant uses interpreted from a perspective of a biological response can reveal different phylogenetic patterns of presumed underlying bioactivity compared to standardised methods of medicinal plant use classification. In the cosmopolitan and pharmaceutically highly relevant genus Euphorbia L., identifying plant uses modulating the inflammatory response highlighted a greater phylogenetic diversity and number of potentially promising species than standardised categories. Our interpretation of medicinal plant uses may therefore allow for a more targeted approach for future phylogeny-guided drug discovery at an early screening stage, which will likely result in higher discovery rates of novel chemistry with functional biological activity.

Phytochemical relationship of Euphorbia helioscopia and Euphorbia pulcherrima with Lactuca sativa.

Allelopathy is an important phenomenon that modifies the ecosystem. A plant can enhance or reduce the growth of other plant due to the presence of a number of allelochemicals in its different parts. Euphorbia helioscopia and Euphorbia pulcherrima are medicinal plant species. Both these species are collected from wild resources for various purposes. To reduce the pressure on wild population, it is important to bring them into cultivation. Therefore, the allelopathic effects of E. helioscopia and E. pulcherrima on the growth of lettuce seeds were studied. Three different concentrations (2%, 4% and 6%) of five different solvents (methanol, acetone, ethyl acetate, n-hexane and distilled water) were used to estimate the allelopathic potential of the above-mentioned Euphorbia species. Results indicated a non-significant growth inhibitory effect of both plants on lettuce seeds. Different extracts reduced the growth of test plant to some extent but this inhibition was not significant. From the observed results, it was concluded that the studied Euphorbia species, being medicinally important crops, can be introduced as intercrop with other cash crops.

[Identification of medicinal plants used as Tibetan traditional medicine Chuan-Bu based on nrDNA markers].

OBJECTIVE: To identify the common Tibetan herb Chuan-Bu. METHOD: Local herbalists were visited to observe which plants were being used as Chuan-Bu. Samples of the indigenous plants were collected at the same time. Leaf materials were collected from field surveys. Total genomic DNA was extracted from silica gel-dried leaf samples. The PCR products were purified and directly sequenced. RESULT: As the origin of Chuan-Bu in Tibet autonomous region was authenticated, two species were determined, i. e. Euphorbia stracheyiand E. wallichii. Also, based on our earlier research, the origin of Chuan-Bu in Gansu province, is from E. kansuensis. The sequences of ITS1 for E. stracheyi and E. wallichii were 261 bp in size, and 221 bp in ITS2, respectively. The size of the 5.8S coding region was 164 bp for all species examined in the genus. Especially, there was a heterozygous locus in ITS1 (C/G; position 72) for E. stracheyi. The nucleotide divergence between sequences of the 6 species in pairwise comparisons was calculated and the result showed that the variable site could be detected in each pairwise comparison of sequences. Also, there were 8 point mutations in the 5.8S coding region. CONCLUSION: nrDNA ITS sequences can be used as the molecular markers to identify the Tibetan herb Chuan-Bu and such Traditional Chinese Medicines from the same genus Euphorbia as E. lathyris, E. humifusa and E. pekinensis.

Differentiation of the traditional Chinese medicinal plants Euphorbia humifusa and E. maculata from adulterants by TaqMan real-time polymerase chain reaction.

DNA sequence analysis of the rDNA internal transcribed spacer 1 (ITS1) and TaqMan real-time polymerase chain reaction were exploited for their applications in the differentiation of the traditional chinese medicinal plants euphorbia humifusa and e. maculata from three related adulterants e. hypericifolia, E. atoto and E. prostrata. The data demonstrated that variations in the ITS1 regions were very low at the intra-species level but extremely high at the inter-species level, so that they could be easily distinguished at the DNA level. The sequence difference allowed an effective and reliable differentiation of E. humifusa and E. maculata from the adulterants by TaqMan real-time PCR.

Authentication of the 31 species of Toxic and Potent Chinese Materia Medica (T/PCMM) by microscopic technique, part 2: Three species of seed T/PCMM.

Toxic and Potent Chinese Materia Medica (T/PCMM) are being used more and more in the treatment of various diseases. In view of their toxic side effects and to ensure their safe use, accurate and reliable authentication is indispensable. However, identifying characteristics of T/PCMM are seldom reported, even though modern microscopy can provide ample, unique identifying characteristics from cells found in transverse sections and powders. In particular, no systematic authentication studies on seed T/PCMM have been conducted. In the course of our study on 31 T/PCMM originating from plants, animals, minerals, and secreta, an accurate and convenient method, based on microscopic techniques, has been developed and reported for the authentication of animal T/PCMM. The present study deals with detailed investigations on three species of seed T/PCMM, namely Semen Hyoscyami (Hyoscyamus niger L.), Semen Euphorbiae (Euphorbia lathyris L.), and Semen Strychni (Strychnos nux-vomica L.). The macroscopic characters are here described in detail, and the microscopic characters were conclusively determined by common and polarized light microscopy. Results showed that these three T/PCMM can be easily identified by the present method even when powdered and combined. Thus, the microscopic method is applicable for authentication of the earlier three T/PCMM, and the morphological and microscopic characteristics described here are proposed as parameters to establish the authenticity of these three T/PCMM.

Screening of antioxidant activity of three Euphorbia species from Turkey.

Euphorbia acanthothamnos, E. macroclada and E. rigida were investigated for their antioxidant activity. The antioxidant potential of extracts of E. acanthothamnos, E. macroclada and E. rigida was evaluated using different complementary antioxidant tests.

Molecular authentication of the traditional Chinese medicinal plant Euphorbia pekinensis.

The ITS regions of Euphorbia pekinensis and six other Euphorbia species used as adulterants of E. pekinensis were sequenced to differentiate them. The sequences are identical among the individuals in the seven species studies. Diversity in DNA sequences among various species was found ranging from 8.3% to 43.8% in ITS1 and 7.6% to 36.6% in ITS2 region. Furthermore, based on the divergent ITS regions, species-specific primers, JDJp 1 and JDJp 2, were designed in the polymorphic regions of E. pekinensis to distinguish it from adulterants. These ITS-derived primers amplified a 281-bp-specific DNA fragment from E. pekinensis. No amplified product was observed using DNA of six adulterants.

The molluscicidal activities of some Euphorbia milii hybrids against the snail Indoplanorbis exustus.

The objective of this study was to observe the molluscicidal activities of Euphorbia milli, known as "poysean" in Thailand, against Indoplanorbis exustus. Latex from 12 different E. milii hybrids was screened for its molluscicidal activities. Indoplanorbis exustus were exposed for 24 and 48 hours to the latex at various concentrations ranging from 6 to 25 ppm and mortality rates were recorded. Eight hybrids of latex were effective. The six most effective hybrids were E. milii Dang-udom, E. milii Arunroong, E. milii Raweechotchuong, E. milii Srisompote, E. milii Sri-umporn and E. milii Tongnopakun, which killed all snails after 24 hours of exposure. Under the same conditions, latex of E. milii Dowpraket and E. milii Promsatid killed 50% of the snails. Such results indicate that these 6 hybrids seem promising as natural molluscicidal agents.