Plant Euphorbia Research Articles

Extraction, Phytochemical Screening, Isolation and Identification of Bioactive Compounds from Extract of the plant Euphorbia Thymifolia Linn

Medicinal plants play an important role in the development of potent therapeutic agents. Plant based drugs provide outstanding contribution to modern therapeutics as a source of many valuable secondary metabolites which serves as plant defense mechanisms against predator such as microorganism, insects and herbivores which have been proved to be potentially active compounds. Euphorbia Thymifolia Linn (E. Thymifolia) is commonly known as ‘duddi’ or in Sanskrit means Laghu didhika or Raktavindaka. It belongs to the family Euphorbiceae. This plant is bitter, acrid, sweet and used as thermogenic, laxative and diuretic. This plant is widely used in the ayurveda to cure many diseases like vitiated condition of constipation, helminthiasis and ringworm skin diseases and leprosy. The aim of the present study is to examine E.Thymifolia Linn whole plant for phytochemical profile, Isolation and Identification of bioactive compounds. Qualitative analysis of various phytochemical constituents was determined by the well-known test protocol available in the literature. Isolation and characterization of bioactive compound from methanolic extract of E. Thymifolia has been conducted. The bioactive compound from methanolic extracts was isolated by several processes, such as TLC, column chromatography and preparative TLC. The isolated bioactive compound is identified by UV-Vis spectrophotometer, FT-IR, 1H, 13C-NMR and Mass. The obtained compound is continued to the preparative TLC using chloroform: methanol (50:50, v/v) as eluent. The UV-Vis spectrum showed one peaks of maximum absorbance at 312.8nm. Then, the FT-IR spectrum showed several peaks that confirmed the presence of functional group of derivative of compound, i.e. 669.05, 928.58, 1070.85, 1215.51 and 1710.07cm-1. 1H and 13C-NMR spectrum confirmed the bioactive compound present in plant. Phytochemical analysis revealed the presence of alkaloids, glycosides, phenols, flavonoids, tannins. The findings of the present study will be helpful to phytochemists, pharmacologists and pharmaceutical industries.
 Keywords: Euphorbia Thymifolia, Qualitative phytochemical, Isolation, Bioactive compounds

Chemical constituents from Euphorbia stracheyi Boiss

The chemical investigation of whole plants Euphorbia stracheyi Boiss. led to the isolation of 14 compounds, including eight diterpenes (1–8), one monoterpene (9), three coumarins (10–12), and two phenols (13–14). Their structures were elucidated by extensive spectroscopic analyses and by comparison with the literature. Compounds 1–6, and 8–12 were firstly isolated from E. stracheyi, while compounds 6, and 9 were isolated from Euphorbia genus for the first time. The chemotaxonomic significance of these isolated compounds is discussed.

The Isolation and Characterization of Phenolic Compound of Euphorbia Plant/Patikan Cina (Euphorbia thymifolia Linn)

Euphorbia plant, Euphorbia thymifolia Linn, is one the Euphorbiaceae families that still need to be developed due to its benefits. An effort to be able to use this plant maximumly is by studying the active component in the plant. This study aims to isolate and characterize phenolic compound in euphorbia plant extract using ethyl acetate. 500 gram of euphorbia plant (Patikan Cina) powder was macerated using methanol. Then, the macerated extract was evaporated to eliminate excess solvent and as a result, a solvent-free extract was obtained. Next, the result was partitioned with a solution of methanol : ethyl acetate (1:1) in order to get methanol and ethyl acetate extracts. The ethyl acetate extract of the partition was evaporated to get concentrated ethyl acetate extract. Next, it was isolated in a vacuum liquid and gravitational column chromatography to get pure isolate. Silica gel 60 (0.040 – 0.063 mm) and n-hexane motion phase were used in the separation of concentrated ethyl acetate in vacuum liquid chromatography. There were 16 fractions produced from the yield. Then the fractions were put in TLC. With silica gel 60 (0.2 – 0.5 mm) mesh and n-hexane mobile phase, ethyl acetate gave 5 fractions in gravitational column chromatography and fraction 3 produced a single spot. UV and IR spectroscopy were used to determine the constituents in the isolate. From UV spectrum, max is 268.97 nm. While IR data shows the presence O-H group in 3521.38 cm-1, C – H (alkane) presents at wave number 2926.45 cm-1 reinforced by the appearance of 5 other alkanes absorption at 2857.00 cm-1, 1447.31 cm-1, 1370.18 cm-1, 868.774 cm-1 and 757.887 cm-1 wavelengths. There is also C = O (carbonyl) groups in wave number 1693.19 cm-1. The identification of structures based on UV and IR spectra data has shown that the isolate is a phenolic compound.

Chemical ecology traits in an adaptive radiation: TPA-sensitivity and detoxification in Hyles and Hippotion (Sphingidae, Lepidoptera) larvae

The larvae of several species in the hawk moth genus Hyles, including H. euphorbiae, feed on plants of the genus Euphorbia containing phorbol esters and are insensitive to addition of the standard phorbol ester, tetradecanoyl-phorbol-13-acetate (TPA) to their artificial diet. Specialised non-Euphorbia feeding larvae were sensitive, whereas polyphagous ones were insensitive if their natural food plant spectrum also includes Euphorbia. Larvae of Hippotion celerio, an out-group species with polyphagous larvae not using Euphorbia as food plants, were sensitive. A highly conserved sequence of the TPA binding site of the protein kinase C in H. euphorbiae and Hippotion celerio demonstrates that intoxication by phorbol esters is not avoided by preventing target binding. H. euphorbiae larvae showed no vitality loss after chemical destruction of their peritrophic matrix and subsequent TPA treatment. TPA fed larvae that had putatively piperonyl butoxide (PBO)-inhibited cytochrome P450 enzymes also showed no deficits, indicating that this is not the only detoxification pathway in H. euphorbiae. Based on these qualitative results, we postulate that proto-Hyles was polyphagous with the ability to use Euphorbia as food plants. The most ancestral Hyles species presumably remained polyphagous, with the ability to switch to the toxic food plants if necessary. Within the youngest, Palearctic radiation, the species specialised on non-Euphorbia plants subsequently lost detoxification abilities, albeit not to 100%. The species in South America, the origin of Hyles, can detoxify TPA, indicating this ability to be a plesiomorphic character state within the genus that enabled its adaptive radiation in the Palearctic, the distribution and diversity centre of Euphorbia.

Endophytic FungusAspergillus japonicusMediates Host Plant Growth under Normal and Heat Stress Conditions

We have isolated an endophytic fungus with heat stress alleviation potential from wild plant Euphorbia indica L. The phylogenetic analysis and 18S rDNA sequence homology revealed that the designated isolate was Aspergillus japonicus EuR-26. Analysis of A. japonicus culture filtrate displayed higher concentrations of salicylic acid (SA), indoleacetic acid (IAA), flavonoids, and phenolics. Furthermore, A. japonicus association with soybean and sunflower had improved plant biomass and other growth features under high temperature stress (40°C) in comparison to endophyte-free plants. In fact, endophytic association mitigated heat stress by negotiating the activities of abscisic acid, catalase, and ascorbic acid oxidase in both soybean and sunflower. The nutritional quality (phenolic, flavonoids, soluble sugars, proteins, and lipids) of the A. japonicus-associated seedlings has also improved under heat stress in comparison to endophyte-free plants. From the results, it is concluded that A. japonicus can modulate host plants growth under heat stress and can be used as thermal stress alleviator in arid and semiarid regions of the globe (where mean summer temperature exceeds 40°C) to sustain agriculture.

In vitro evaluation of antiurolithiatic activity of Euphorbia thymifolia L. plant extracts

Objectives: The plant Euphorbia thymifolia Linn., Family: Euphorbiaceae, is used in traditional medical practices of India to treat various diseases. It was suspected and found to evaluate the effect of E. thymifolia plant extracts for in vitro antiurolithic activity on generated calcium oxalate crystals.Method: The aqueous, hydroalcoholic, and alcoholic (ethanolic) extracts of plant were tested for antiurolithiatic potential on generated calcium oxalate crystals by homogeneous precipitation method, and simultaneously, supporting two-step vice versa reactions were assessed (new method). The activity was assessed by studying the crystal dissolution by microscopy and quantitative alimental ions analysis for calcium and oxalates.Results: They exhibited significant activity when compared to standard drug cystone-polyherbal formulation. The aqueous and alcoholic extracts significantly decreased (P < 0.001) crystal size and increased calcium and oxalate concentration in reaction setup of all tested groups as compared to normal control. Simultaneously, a supporting two-step vice versa reaction was assessed which have shown a significant inhibition of crystal formation.Conclusion: All the interpretations of various result outcomes direct the use of this drug for urolithiasis prophylactic as well as curative treatments.

Bioenergy potentials of three Euphorbia species from Western Ghats in India

ABSTRACTBioenergy constituents of three Euphorbia species viz., Euphorbia vajravelui, E. tortilis, and E. trigona, were evaluated. These Euphorbia species showed moisture (79.5–91.9%), ash (4.06–6.80%), latex (0.08–0.26 g), oil (5.45–5.59%), polyphenol (3.12–8.39%), hydrocarbon (0.28–0.41%), carbon (42.40–44.5%), hydrogen (6.22–8.40%) contents and gross heat values (17.51–18.71 MJ/kg) comparable to other reported Euphorbia plants, energy crops and biomass sources. Thermal analyses (TG/DTG, DTA, DSC) produced comparable results. Trace elements such as Fe, Mg, K, Ca, P, Zn, Mn, Ba, Sr, Cu, Sc, and As were detected in these Euphorbia specimens. This study reveals the biomass/bioenergy potentials of these three Euphorbia species from Western Ghats, India.

Euphornin reduces proliferation of human cervical adenocarcinoma HeLa cells through induction of apoptosis and G2/M cell cycle arrest.

BackgroundThe plant Euphorbia helioscopia L. has been used in traditional Chinese medicine for treating various disorders such as tuberculosis and edema. The aim of this study was to investigate the effect of euphornin, a bioactive compound isolated from E. helioscopia, on proliferation of human cervical adenocarcinoma HeLa cells by analyzing cell viability, rate of apoptosis, and cell cycle progression.Materials and methodsThe sulforhodamine B assay was used to study the effect of euphornin on the proliferation of HeLa cells. Morphological changes to cell nuclei were identified after Hoechst 33342 staining. Mitochondrial membrane depolarization (MMP) was analyzed after staining with JC-1 dye. The influence of euphornin on the apoptosis rate was analyzed by Annexin V/propidium iodide double staining. Fluorescence-activated cell sorting was applied to investigate the influence of euphornin on cell cycle progression. Proteins were obtained from HeLa cells and analyzed by Western blots.ResultsA cell viability assay showed that euphornin inhibited proliferation of HeLa cells in a dose-dependent and time-dependent manner. Euphornin also induced apoptosis in a concentration-dependent manner, with the rates of apoptosis ranging from 25.3% to 52.6%. A high concentration of euphornin was found to block HeLa cells at the G2/M stage. A Western blot analysis suggested that euphornin might exhibit antitumor activity by inducing apoptosis. Euphornin treatment altered the ratio of Bax/Bcl-2 in HeLa cells, which led to the release of cytochrome complex. The levels of cleaved caspase-3, caspase-8, caspase-9, and caspase-10 were also markedly increased by euphornin treatment. Analysis of cell cycles indicated that euphornin induced cell cycle arrest by increasing the level of the phospho-CDK1 (Tyr15) protein. The various assays demonstrated that euphornin treatment resulted in a significant suppression of cell growth accompanied by G2/M cell cycle arrest and increased rate of apoptosis via mitochondrial and caspase pathways.ConclusionOur findings suggest that euphornin has the potential to be used as a cancer therapeutic agent against human cervical adenocarcinoma.

Euphol, a tetracyclic triterpene, from Euphorbia tirucalli induces autophagy and sensitizes temozolomide cytotoxicity on glioblastoma cells.

Glioblastoma (GBM) is the most frequent and aggressive type of brain tumor. There are limited therapeutic options for GBM so that new and effective agents are urgently needed. Euphol is a tetracyclic triterpene alcohol, and it is the main constituent of the sap of the medicinal plant Euphorbia tirucalli. We previously identified anti-cancer activity in euphol based on the cytotoxicity screening of 73 human cancer cells. We now expand the toxicological screening of the inhibitory effect and bioactivity of euphol using two additional glioma primary cultures. Euphol exposure showed similar cytotoxicity against primary glioma cultures compared to commercial glioma cells. Euphol has concentration-dependent cytotoxic effects on cancer cell lines, with more than a five-fold difference in the IC50 values in some cell lines. Euphol treatment had a higher selective cytotoxicity index (0.64-3.36) than temozolomide (0.11-1.13) and reduced both proliferation and cell motility. However, no effect was found on cell cycle distribution, invasion and colony formation. Importantly, the expression of the autophagy-associated protein LC3-II and acidic vesicular organelle formation were markedly increased, with Bafilomycin A1 potentiating cytotoxicity. Finally, euphol also exhibited antitumoral and antiangiogenic activity in vivo, using the chicken chorioallantoic membrane assay, with synergistic temozolomide interactions in most cell lines. In conclusion, euphol exerted in vitro and in vivo cytotoxicity against glioma cells, through several cancer pathways, including the activation of autophagy-associated cell death. These findings provide experimental support for further development of euphol as a novel therapeutic agent for GBM, either alone or in combination chemotherapy.

New cycloartane-type ester triterpenes from Euphorbia pterococca and biological evaluation

From acetonic extract of the whole plant Euphorbia pterococca Brot. (Euphorbiaceae), four new cycloartane-type ester triterpenes named cycloartenyl-2′E,4′E-decadienoate (1), cycloartenyl-2′E,4′Z-decadienoate (2), 24-methylenecycloartanyl-2′E,4′Z-tetradecadienoate (3), and 24-oxo-29-norcycloartanyl-2′E,4′Z-hexadecadienoate (4) were obtained along with nine known tetracyclic triterpenes (5–13). Their structures were established mainly by extensive use of spectroscopic techniques, including 1D (1H and 13C) and 2D homo- and heteronuclear NMR experiments (COSY, HSQC, HMBC and NOESY), and mass spectrometry (HRESIMS), and by comparison with data reported in the literature. In addition, the new compounds 1–3 have been tested for cytotoxicity, trypanocidal effects and on enzymes involved in endocannabinoid degradation. While inactive in all assays up to 100 μM, 1 showed selective inhibition of α/β-hydrolase 12 with an IC50 of 11.6 ± 1.9 μM.

Full extract of Euphorbia esula reversed chemoresistance, inhibited cell migration/invasion, and induced apoptosis of multidrug-resistant SGC7901/VCR cells

Background: The plant Euphorbia esula has been used by ancient Chinese people to treat cancer and other ailments and is used by present doctors or folks as an assistant treatment for some kind of cancers, but the mode of action is unclear. Objective: To investigate the influence of E. esula full extract on chemoresistance, cell migration/invasion, and apoptosis of multidrug-resistant human stomach cancer cells. Materials and Methods: After treating multidrug-resistant human stomach cancer SGC7901/VCR cells with E. esula extract at varying concentrations, the inhibition of cell proliferation was detected using thiazolyl blue. Sensitivity to the chemotherapeutic drugs, adriamycin and paclitaxel, was evaluated using half maximal inhibitory concentration. Cell cycle progression was analyzed by flow cytometry. The inhibitions of cell migration and invasion were examined by Transwell method. The induction of apoptosis and the apoptotic rate were studied by electron microscopy and flow cytometry, respectively. Activation of caspase 3 enzyme was inspected by ultraviolet spectrophotometry. Results: E. esula extract could increase the sensitivity of SGC7901/VCR cells to the chemotherapeutic drugs, adriamycin and paclitaxel. The proliferation, migration, and invasion of SGC7901/VCR cells were significantly inhibited by E. esula extract (P < 0.01 compared with negative control), which showed time- and dose-dependent manners (P < 0.05 and P < 0.01, respectively). Cell cycle was arrest at the S phase. E. esula extract also induced apoptosis in SGC7901/VCR cells, and the apoptotic rate was increased significantly with drug concentration and with treatment time (P < 0.01 compared with negative control, P < 0.05 between concentrations and time points). E. esula extract upregulated enzymatic activity of caspase 3. Conclusion: E. esula extract could reverse SGC7901/VCR cell's resistance to chemotherapeutic drugs and could inhibit cell proliferation, migration, and invasion, interfere with cell cycle progression, and induce apoptosis in SGC7901/VCR cells. Abbreviations used: E. esula: Euphorbia Esula; FCS: fetal calf serum; IC50: half maximal inhibitory concentration; OD: optical density.

Phytochemical and Antioxidant Activities of Methanolic Extract of the Plant Euphorbia thymifolia Linn

The present study was designed for phytochemical and antioxidant activity of methanolic extract of the whole plant Euphorbia thymifolia linn. Belonging to the family Euphorbiaceae. It cures many diseases. The phytochemicals present in this methanolic extract were studied by standard Protocols and the invitro antioxidant activities were studied using different methods i.e., H2O2 scavenging activity, Reducing power method and Phosphomolybdinum method. These three methods show significant activity with standard drugs.

Evaluation of genotoxicity of Euphorbia triaculeata Forssk. extract on mice bone marrow cells in vivo.

Chemotherapeutic agents for cancer are highly toxic to healthy tissues at therapeutic doses and hence alternative medicine avenues are widely researched. Most of the studies on alternative medicine have suggested that Euphorbia plant possesses considerable antitumor and antibacterial properties. The present study was designed to evaluate the in vivo genotoxic effects of Euphorbia triaculeata extract on mice bone marrow cells using chromosomal aberration test and micronucleus assay. This study also deals with the effect of E. triaculeata on the standard drug cyclophosphamide (CP) treatment in mice. Three different doses 250, 500 and 1000 mg/kg body weight were selected. In micronucleus assay, single oral dose administration of Euphorbia triaculeata extract at the three doses did not increase the number of micronucleated polychromatic erythrocytes. Similarly, a single oral administration of Euphorbia triaculeata extract showed no significant changes on mitotic indices or in induction of chromosomal aberrations in mice bone marrow cells. Pretreatment with E. triaculeata extract significantly reduced the clastogenicity of CP. Hence it can be concluded that, E. triaculeata extract showed no significant genotoxic effect on mice bone marrow cells. Under the conditions of this study, it has been demonstrated that the Euphorbia triaculeata extract is not genotoxic and not clastogenic at the concentrations used.

Apoptosis-inducing Effect of the Hexane Extracts from Three Native Iranian Euphorbia Plants

Background: Cancer is one of the most significant causes of death. Plants with anti-cancer effects have shown to eliminate tumor cells by the induction of apoptosis. Objectives: The present study aimed at investigating the apoptotic effects of 3 Euphorbia native species (E. microciadia Boiss, E. osyridea Boiss, and E. heteradenia Jaub. & Sp.). Methods: Cell lines with the strongest sensitivity to the extracts including HeLa for E. microciadia, K562 for E. heteradenia, and Fen for E. osyridea were investigated for the effects of the plants’ hexane extracts on the induction of apoptosis, according to annexin V/propidium iodide staining by flow cytometry. We used real-time PCR to evaluate the changes in expressions of genes related to the intrinsic and extrinsic apoptosis pathways. Results: Flow cytometry results indicated that 50 μg/mL of all the extracts induced apoptosis in more than 60% of treated cells. Administration of the extracts resulted in increased caspase-3 activity for all treated cell lines. Real-time PCR results showed decreased B-cell lymphoma-2 (Bcl-2) and increased Bax and Fas mRNA levels in cells treated with the extracts. The Bax/Bcl-2 ratio in cells treated with 50μg/mL of E. microciadia was 29.25 ± 12.0 (P < 0.05) and for 100 μg/mL of E. heteradenia. and E. osyridea was 30.2 ± 13.0 (P < 0.05) and 22.2 ± 2.4 (P < 0.001), respectively. Conclusions: The studied plants have shown remarkable apoptosis-inducing effects on tumor cell lines by affecting both the extrinsic and intrinsic apoptosis pathways. Additional studies in terms of their beneficial effects as natural sources of potential anti-cancer agents will be necessary.

Attenuation of nociceptive pain and inflammatory disorders by total steroid and terpenoid fraction of Euphorbia tirucalli Linn root in experimental in vitro and in vivo model.

The plant Euphorbia tirucalli Linn has been successfully used as a tribal folk medicine in India and Africa for the management of acute inflammatory, arthritic, nociceptive pain and asthmatic symptoms. The present study was conducted to assess the anti-inflammatory, analgesic, anti-asthmatic and anti-arthritic role of the total steroid and terpenoid rich fractions of the hydro-alcoholic extract of E. tirucalli root (STF-HAETR). STF-HAETR fraction demonstrated 71.25±2.5 and 74.25±5.1% protection against acetic acid-induced pain and central neuropathic pain at 75 and 100mg/kg doses, respectively. It showed 96.97% protection against acute inflammation at 100mg/kg with 1.6-fold better activity than the standard drug. The fraction exhibited such efficacy via inhibition of proinflammatory cytokines TNF-α, IFN-γ, by 61.12 and 65.18%, respectively, at 100μg/mL. Inhibition of cyclooxygenase and Nitric oxide synthase in a dose-dependent manner affirms its analgesic and anti-inflammatory activity. The spectrophotometric analysis reveals that STF-HAETR induces ameliorative effect against heat-induced denaturation of Bovine serum albumin (BSA) and exhibits significant anti-proteinase activity. The plant fraction also demonstrated anti-asthmatic activity by displaying 62.45% protection against histamine induced bronchoconstriction or dyspnoea. Our findings suggest that STF-HAETR could be an effective safe therapeutic agent to treat nociceptive pain, acute inflammation, asthma, and arthritis which may authenticate its traditional use.

Influence of valoneoyl groups on the interactions between Euphorbia tannins and human serum albumin

Tannins belonging to plant polyphenols, are large group of compounds with diverse biological activity. Many of them are being studied as potential natural medicine due to their antioxidant, antiviral, antibacterial or anticancer properties. However, so far little is known about the structural and functional relations in protein-tannin interactions, in particularly the role of valoneoyl groups in tannin structure. In this study we first investigated the mechanisms of interaction 1,2-di-O-galloyl-4,6-valoneoyl-β-D-glucose (Tannin 1), 2-O-galloyl-4,6-valoneoyl-β-D-glucose (Tannin 2), 3-O-galloyl-1,2-valoneoyl-β-D-glucose (Tannin 3) isolated from Euphorbia plants with human serum albumin (HSA). To get more detailed information about nature of albumin-tannin interactions besides standard Trp fluorescence quenching analysis we used also transmission electron microscopy (TEM), circular dichroism (CD) and fluorescence labelling (ANS dye) techniques. It was shown that all the tannins strongly interacted with HSA and quenched the tryptophan amino acid (Trp) fluorescence but slightly changed protein secondary structure (circular dichroism CD analysis). TEM demonstrated that all used compounds formed complexes with HSA. Tannin 3 most strongly quenched HSA fluorescence and changed protein dynamic as well as had the highest binding constant (12.4 ± 1.1 × 1013M−1s−1 in comparison with 7.0 ± 0.38 × 1013M−1s−1 for tannin 1 and 8.6 ± 1.1 × 1013M−1s−1 for tannin 2). For tannin 1 that was the largest of the studied compounds was observed the weakest influence on the fluorescence parameters, probably due to the effect of steric hindrance limiting interaction with albumin Trp pocket. On the other hand T1 induced the strongest changed secondary structure of HSA in comparison to another studied tannins.Our results demonstrated that all used tannins interact with albumin by complex formation but in the manner depends on chemical structure and flexibility of studied compounds.

Euphorbia lunulata extract acts on multidrug resistant gastric cancer cells to inhibit cell proliferation, migration and invasion, arrest cell cycle progression, and induce apoptosis

Ethnopharmacological relevanceThe milky sap or the aboveground part of the plant Euphorbia lunulata has long been used by Chinese people to treat noncancerous growths and cancerous ailments but the specific mode of action and the action mechanism remain to be elucidated. Aim of the studyTo investigate the effects of Euphorbia lunulata extract on cell proliferation, migration, invasion, cell cycle progression, and apoptosis of multidrug resistant human gastric cancer cells; To study the mechanism of apoptosis induction by Euphorbia lunulata extract in multidrug resistant human gastric cancer cells. Materials and methodsThe aboveground part of fresh Euphorbia lunulata plant was extracted first with ethanol and then with n-hexane. The aseptic extract at varying concentrations was used to treat the multidrug resistant human gastric cancer SGC7901/ADR cells. After treatment, the inhibition of cell proliferation was examined by MTT assay. The inhibitions of cell migration and invasion were detected by Transwell method. The alteration of cell cycle progression was studied by flow cytometry. The morphological changes of cell nuclei were observed with fluorescence microscopy following Hoechst 33258 staining and the apoptotic indexes were calculated. The activation of caspase enzymes was analyzed by spectrophotometry. The sub-cellular distribution of cytochrome complex and the expression of Bax and Bcl-2 proteins were determined by Western blot. ResultsThe proliferation, migration, and invasion of SGC7901/ADR cells were significantly inhibited by Euphorbia lunulata extract, which showed time- and dose-dependent manners. Cell cycle was arrested in G2/M phase. Significant apoptotic morphological changes were observed in the nuclei of the treated cells, and apoptotic indexes were increased significantly; these changes were diminished when Z-VAD-FMK, a caspase inhibitor, was also presented. The activities of caspase-3, caspase-8, and caspase-9 were increased. The sub-cellular distribution of cytochrome complex was altered----reduced in the mitochondria and increased in the cytoplasm. The expression of Bax was upregulated, while that of Bcl-2 was downregulated. ConclusionEuphorbia lunulata extract inhibited the proliferation, migration, and invasion of SGC7901/ADR cells, arrested cell cycle progression, and induced cell apoptosis; the mechanism of apoptosis induction involved both the extrinsic and the intrinsic pathways.

Quantitative Analysis of Ingenol in Euphorbia species via Validated Isotope Dilution Ultra-high Performance Liquid Chromatography Tandem Mass Spectrometry.

Various species of the Euphorbia genus contain diterpene ingenol and ingenol mebutate (ingenol-3-angelate), a substance found in the sap of the plant Euphorbia peplus and an inducer of cell death. A gel formulation of the drug has been approved by the US Food and Drug Administration (FDA) and the European Medicines Agency (EMA) for the topical treatment of actinic keratosis. To develop a rapid and reliable method for quantification of ingenol in various plant extracts. Methanolic extracts of 38 species of the Euphorbia genus were analysed via ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) after methanolysis and solid-phase extraction (SPE) purification. The 18 O-labelled ingenol analogue was prepared and used as an internal standard for ingenol content determination and method validation. The highest ingenol concentration (547mg/kg of dry weight) was found in the lower leafless stems of E. myrsinites. The screening confirms a substantial amount of ingenol in species studied previously and furthermore, reveals some new promising candidates. The newly established UHPLC-MS/MS method shows to be an appropriate tool for screening of the Euphorbia genus for ingenol content and allows selection of species suitable for raw material production and/or in vitro culture initiation. Copyright © 2017 John Wiley & Sons, Ltd.

Pathway engineering for the production of β-amyrin and cycloartenol in Escherichia coli-a method to biosynthesize plant-derived triterpene skeletons in E. coli.

Cycloartenol is biosynthetically the first sterol skeleton, which is metabolized to phytosterols such as β-sitosterol and stigmasterol. β-Amyrin is the most commonly occurring aglycone skeleton for oleanane-type saponins such as glycyrrhizin and saikosaponins. It has been regarded that these cyclic triterpenes are unable to be produced in Escherichia coli, while no reports are available on their production with E. coli. Here, we describe a method to synthesize triterpene skeletons from higher plants, including cycloartenol and β-amyrin. We introduced into E. coli the biosynthetic pathway genes from farnesyl diphosphate (FPP) to cycloartenol or β-amyrin, which contained Arabidopsis (Arabidopsis thaliana)-derived squalene synthase (AtSQS) and squalene epoxidase (AtSQE) genes in addition to the Arabidopsis cycloartenol synthase (AtCAS1) gene, or the β-amyrin synthase (EtAS) gene of the petroleum plant Euphorbia tirucalli, along with the isopentenyl diphosphate isomerase (HpIDI) gene from a green algae Haematococcus pluvialis. The order of genes, HpIDI, AtSQS, AtSQE, driven by transcriptional read-through from a tac promoter to an rrnB terminator, was crucial for their functional expression in E. coli to produce cycloartenol or β-amyrin. The co-expression of a bacterial NADPH-regenerating gene (zwf or gdh) as well as bacterial redox partner protein genes (camA and camB, or NsRED and NsFER) was found to increase the amounts of these triterpenes several fold. The present study could open up opportunities not only to carry out functional analysis of a higher-plant-derived oxidosqualene cyclase (OSC) gene in E. coli but also to produce functional triterpenes that originate from medicinal or herbal plants.

Green synthesis of γ-Fe2O3 nanoparticles for methane gas sensing

Green synthesis is becoming increasingly important as an eco-friendly alternative to the traditional production process because of its growing industrial applications. In this study, peroxidase enzymes of the leaf extracts of the plants Ficus carica (Fig) and Euphorbia amygdaloides (Euphorbia) were used for the synthesis of γ-Fe2O3 nanoparticles (NPs). The structural, morphological and methane gas sensing properties of γ-Fe2O3 NPs were investigated. γ-Fe2O3 NPs obtained from the Fig plant peroxidase enzyme showed a higher response, selectivity and short response/recovery time than γ-Fe2O3 NPs obtained from the Euphorbia plant peroxidase enzyme, which exhibited 15% response to 1-ppm methane gas at 150 °C. Impedance spectroscopy analysis showed that the resistance due to the grain boundaries significantly contributed to the characteristics of gas sensing. It can be seen from results that γ-Fe2O3 NPs produced from peroxidase enzymes that purified from Fig plant have great potential for industrial applications.