Chemical Composition of the Essential Oils of the Iberian Peninsula Endemic Species Eryngium dilatatum Lam.

Eryngium dilatatum Lam. is a thorny Iberian Peninsula endemic species belonging to the Apiaceae family that has not been previously analysed from a chemical point of view. Following our studies on this genus, we characterized the chemical composition of the essential oils from the different parts (inflorescences, stems + leaves, and roots) of this species; these parts were gathered in Cádiz (Spain). The specimens were collected in July during the flowering period and air-dried before the oil extraction by hydro-distillation. The essential oils were analysed by gas chromatography and gas chromatography coupled with mass spectrometry. The different parts of the plant yielded low amounts of pale yellow oil, with the roots being the fraction that provided the lowest amount of oil. The chemical characterization of the essential oils showed qualitative and quantitative differences between the fractions examined, but all of them showed the same principal compound, germacrene D (9.1-46.5%). Similarly, all the fractions shared most of their representative constituents, with their percentage compositions being different from one sample to the other: α-cadinol (3.8%), bicyclogermacrene (3.5%), octanal (3.1%), and spathulenol (2.5%) were found in the inflorescences; octanal (8.1%), α-cadinol (3.7%), δ-cadinene (3.6%), (E)-caryophyllene (2.6%), bicyclogermacrene (2.5%), and spathulenol (2.4%) were found in the stems and leaves; and spathulenol (4.6%), α-cadinol (4.4%), khusinol (3.2%), α-muurolol (3.1%), and δ-cadinene (2.6%) were found in the roots. As far as we know, this is the first report about the chemical composition of this endemic species of the Iberian Peninsula. It contributes to the knowledge of this species and to the genus to which it belongs. This species could be considered as a natural source of germacrene D, which is a sesquiterpene hydrocarbon with active properties.

Eryngium billardieri extract affects cardiac gene expression of master regulators of cardiomyaopathy in rats with high fatdiet-induced insulin resistance.

BACKGROUND: For years, numerous studies have focused on identifying approaches to increase insulin sensitivity by modifying the signaling factors. In the present study, we examined the effects of Eryngium billardieri extract, as an anti-diabetic herbal medication, on the heart mRNA level of Akt serine/threonine kinase (Akt), mechanistic target of rapamycin kinase (mTOR), peroxisome proliferator-activated receptor gamma (PPARγ), and Forkhead box o1 (Foxo1) in rats with high-fat diet (HFD)-induced insulin resistance (IR). We also assessed the anti-diabetic effects of E. billardieri extract in rats with insulin resistance. METHODS: Twenty-seven male Wistar rats were divided into two groups. Nine rats were fed a normal diet (control group), and 18 rats were fed an HFD for 13 weeks (HFD group). To confirm the induction of insulin resistance, the oral glucose tolerance test (OGTT) was performed and homeostatic model assessment for insulin resistance (HOMA-IR) was calculated. Then rats with IR were randomly divided into the following groups: the HFD group, which continued an HFD, and the group treated with E. billardieri extract, which received the extract at a concentration of 50 mg/kg for 30 days. On the 30th day, the animals were sacrificed and serum samples were collected for biochemistry analyses. Furthermore, the expression of Akt, mTOR, PPARγ, and Foxo1 was measured in heart tissue using the real-time polymerase chain reaction (PCR) method. RESULTS: Real-time PCR analyses revealed that an HFD can significantly decrease the expression level of Akt, mTOR, and PPARγ in the heart tissue. However, an HFD significantly increased the expression level of Foxo1 in the HFD group compared to the control group (P < 0.05). In addition, our data showed that the administration of E. billardieri extract significantly enhanced the mRNA levels of Akt, PPARγ, and mTOR in the heart tissue compared to the HFD group (P < 0.05), while it significantly decreased the Foxo1 mRNA levels (P < 0.01). CONCLUSION: Given that Akt, mTOR, PPARγ, and Foxo1 are critical factors in insulin resistance, the present study suggests that E. billardieri could probably be used as an alternative treatment for IR as a major feature of metabolic syndrome.

Phytochemical Diversity Comparison in Leaves and Roots of Wild and Micropropagated Latvian Sea Holly (Eryngium maritimum L.).

The goal of the current study was to compare the chemical composition of the roots, shoots, and leaves of wild-growing Eryngium maritimum L., and of in vitro and in field-cultivated plants in Latvia. The essential oil yield obtained by hydrodistillation ranged from 0.14% to 0.54%, while analysis of the chemical composition using GC-MS revealed a total of 44 different volatiles, with differences in the types and amounts of volatiles between the leaves and roots. Using 96-well plate techniques, the concentration of total phenolic compounds, saponins, and sugars in the aqueous ethanolic extracts of E. maritimum were assessed, along with their capacity to scavenge stable DPPH radicals. Extracts from roots had a lower concentration of total phenolic compounds compared to those from the leaves of wild grown and cultivated plants but did not differ from in vitro shoots. Root, leaf, and shoot samples of the same genotype from different growth conditions had approximately the same concentration of total saponins, while total sugar concentrations were higher in the roots. The growth conditions had a significant effect on the concentration of total phenolic compounds and antiradical activity, with differences that were significant observed between plant aboveground and belowground parts. Analysis using UHPLC-ESI-q-TOF-MS revealed 63 compounds, with amino acids and hydroxycinnamic acid derivatives (such as chlorogenic and rosmarinic acid) being the major compound groups that significantly differed between plant growth conditions. We also demonstrated that rapid screening of volatile compounds in in vitro plants using headspace gas chromatography mass spectrometry analyses can predict the formation of marker compounds in the same mericlones grown in field conditions. These findings provide valuable insights into the chemical composition of E. maritimum and its potential for use in various applications.

Ethnobotany, Biological Activities and Phytochemical Compounds of Some Species of the Genus Eryngium (Apiaceae), from the Central-Western Region of Mexico.

There are approximately 250 species of Eryngium L. distributed throughout the world, with North America and South America being centers of diversity on this continent. In the central-western region of Mexico there may be around 28 species of this genus. Some Eryngium species are cultivated as leafy vegetables, ornamental, and medicinal plants. In traditional medicine they are used to treat respiratory and gastrointestinal conditions, diabetes, and dyslipidemia, among others. This review addresses the phytochemistry and biological activities, as well as traditional uses, distribution, and characteristics of the eight species of Eryngium reported as medicinal in the central-western region of Mexico: E. cymosum, E. longifolium, E. fluitans (or mexicanum), E. beecheyanum, E. carlinae, E. comosum, E. heterophyllum, and E. nasturtiifolium. The extracts of the different Eryngium spp. have shown biological activities such as hypoglycemic, hypocholesterolemic, renoprotective, anti-inflammatory, antibacterial, and antioxidant, among others. E. carlinae is the most studied species, and phytochemical analyses, performed mainly by high-performance liquid chromatography (HPLC) and gas chromatography coupled with mass spectrometry (GC-MS), have shown its content of terpenoids, fatty acids, organic acids, phenolic acids, flavonoids, sterols, saccharides, polyalcohols, and aromatic and aliphatic aldehydes. According to the results of this review on Eryngium spp., they constitute a relevant alternative as a source of bioactive compounds for pharmaceutical, food, and other industries. However, there is a lot of research to be conducted regarding phytochemistry, biological activities, cultivation, and propagation, in those species with few or no reports.

Antioxidant effects of silver nanoparticles obtained by green synthesis from the aqueous extract of Eryngium carlinae on the brain mitochondria of streptozotocin-induced diabetic rats.

Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycemia that affects practically all tissues and organs, being the brain one of most susceptible, due to overproduction of reactive oxygen species induced by diabetes. Eryngium carlinae is a plant used in traditional Mexican medicine to treat diabetes, which has already been experimentally shown have hypoglycemic, antioxidant and hypolipidemic properties. The green synthesis of nanoparticles is a technique that combines plant extracts with metallic nanoparticles, so that the nanoparticles reduce the absorption and distribution time of drugs or compounds, increasing their effectiveness. In this work, the antioxidant effects and mitochondrial function in the brain were evaluated, as well as the hypoglycemic and hypolipidemic effect in serum of both the aqueous extract of the aerial part of E. carlinae, as well as its combination with silver nanoparticles of green synthesis. Administration with both, extract and the combination significantly decreased the production of reactive oxygen species, lipid peroxidation, and restored the activity of superoxide dismutase 2, glutathione peroxidase, and electron transport chain complexes in brain, while that the extract-nanoparticle combination decreased blood glucose and triglyceride levels. The results obtained suggest that both treatments have oxidative activity and restore mitochondrial function in the brain of diabetic rats.

Antioxidant and anti-inflammatory potential of rhizome aqueous extract of sea holly (Eryngium maritimum L.) on Jurkat cells.

ETHNOPHARMACOLOGICAL RELEVANCE: The genus Eryngium is known for producing a wide range of bioactive compounds with proved medicinal properties. In the last years, research has focused on E. maritimum, with previous studies reporting anticancer, antimicrobial, antioxidant, and anti-inflammatory activities. Ethnobotanical literature suggests that it has been traditionally used to treat a wide range of illnesses, having antitussive, diuretic and aphrodisiac properties. Being rhizome one of the most bioactive organs, much of the available references from traditional uses suggest that it has been specifically used to treat renal diseases. In this sense, inflammation and oxidative processes play a major role in kidney dysfunctions, which could be associated to the mechanism of action of the plant extracts. AIM OF THE STUDY: The main aim of the study was to investigate the effects of E. maritimum rhizome extract on the antioxidant and inflammatory response in human immune cells. MATERIAL AND METHODS: Rhizome extracts were obtained from plants growing in Mallorca (Balearic Islands), and its composition was determined using HPLC-DAD, highlighting simple phenolic compounds such as trans-ferulic acid, catechin, chlorogenic acid, epicatechin and rosmarinic acid as the major constituents. Total antioxidant capacity was determined using the FRAP assay. Jurkat cells were cultured to analyse cytotoxicity by cell viability assay. In parallel, cells were stimulated with phytohemagglutinin and treated with different extract concentrations. Gene and protein expression, as well as nitrite and cytokine levels were evaluated as indicators of metabolic responses. RESULTS: The plant extract showed a high diversity of pharmacologically bioactive compounds with potential therapeutic uses. The extract presented null cytotoxicity and exerted antioxidant and anti-inflammatory effects on Jurkat cells by inducing an antioxidant response and reducing cytokine and nitric oxide release and the expression of pro-inflammatory genes. CONCLUSION: The present findings suggest that E. maritimum is a promising phytotherapeutic species because of its strong antioxidant and anti-inflammatory potential, which could explain some of its traditional uses.

Molecular Phylogeny, DNA Barcoding, and ITS2 Secondary Structure Predictions in the Medicinally Important Eryngium Genotypes of East Coast Region of India.

Commercial interest in the culinary herb, Eryngium foetidum L., has increased worldwide due to its typical pungency, similar to coriander or cilantro, with immense pharmaceutical components. The molecular delimitation and taxonomic classification of this lesser-known medicinal plant are restricted to conventional phenotyping and DNA-based marker evaluation, which hinders accurate identification, genetic conservation, and safe utilization. This study focused on species discrimination using DNA sequencing with chloroplast-plastid genes (matK, Kim matK, and rbcL) and the nuclear ITS2 gene in two Eryngium genotypes collected from the east coast region of India. The results revealed that matK discriminated between two genotypes, however, Kim matK, rbcL, and ITS2 identified these genotypes as E. foetidum. The ribosomal nuclear ITS2 region exhibited significant inter- and intra-specific divergence, depicted in the DNA barcodes and the secondary structures derived based on the minimum free energy. Although the efficiency of matK genes is better in species discrimination, ITS2 demonstrated polyphyletic phylogeny, and could be used as a reliable marker for genetic divergence studies understanding the mechanisms of RNA molecules. The results of this study provide insights into the scientific basis of species identification, genetic conservation, and safe utilization of this important medicinal plant species.

Evaluation of anti-proliferative activity of Eryngium caucasicum on melanoma cancer cells.

BACKGROUND: The genus Eryngium is a member of the Apiaceae family that has shown different pharmacological effects mainly including anti-inflammatory, analgesic, anti-cancer, hepatoprotective, and anti-oxidant. Previous research on the anti-cancer activity of Eryngium in some cancer cell lines has led us to explore the anti-proliferative activity of E. caucasicum in the B16F10 cell line. OBJECTIVE: In this study, the antiproliferative activity of E. caucasicum on melanoma cancer cells (B16F10) and non-cancerous cells (HFFF2) were evaluated in vitro. METHODS: The dried plant sample of E. caucasicum was extracted by Soxhlet apparatus with n-Hexane, dichloromethane, and methanol solvents. The effects of cytotoxicity of the extracts by the MTT method on melanoma cancer cells (B16F10) and noncancerous cells (HFFF2) was investigated for 24 and 48 hours. Then, the cytotoxicity of different fractions of the strong extract against normal and cancer cells was evaluated by this method. Annexin V/PI assay was used to study the induction apoptosis via the fractions in cancer cells. FINDINGS: According to the results of the MTT test, n-Hexane extract is the most effective extract against the B16F10 cell line and is a candidate for fractionation with VLC. Among the fractions, 40 and 60% VLC fractions of n-Hexan extract inhibited the growth of B16F10 cells at 24 and 48 hours while, these fractions at IC50 concentration had no cytotoxic effects on normal cells. Treatment of cancer cells with effective extract caused apoptosis and necrosis and 40 and 60% more fractions induced apoptosis in these cells. CONCLUSION: The n-Hexane extract of E. caucasicum and its 40 and 60% fractions showed the highest cytotoxic effect against the B16F10 cell line compared to other extracts and control groups. This inhibition was made through induction of apoptosis.

Anti-hyperglycemic effects of Eryngium billardierei F. Delaroche extract on insulin-resistance HepG2 cells in vitro.

BACKGROUND: Insulin resistance as a major problem is associated with type 2 diabetes mellitus. This study investigated the effect of Eryngium billardierei on insulin-resistance induced HepG2 cells. METHODS AND RESULTS: MTT method was used to evaluate the viability of HepG2 cells treated with various doses of E. billardierei extract. An insulin-resistance model was established in HepG2 cells. Next, MTT assay and Acridine orange staining were performed to investigate the viability of cells in the vicinity of different concentrations of insulin, pioglitazone, and E. billardierei extract in an insulin-resistance media. The glucose uptake test was performed to select the optimal insulin concentration. Expression levels of IR, G6Pase, and PEPCK genes were assessed by real-time RT-PCR. According to obtained data, E. billardierei at concentrations of 0.5 and 1 mg/mL show no toxicity on cells. Furthermore, based on MTT assay and glucose uptake test 10-5 mol/L insulin was chosen as the model group to induce insulin-resistance in HepG2 cells for gene expression analysis. Finally, 1 mg/mL E. billardierei not only induced no cytotoxicity but also showed an increase in the expression of IR as well as a reduction in G6Pase and PEPCK level compared to the control and model groups. CONCLUSIONS: The obtained data indicated that 1 mg/mL E. billardierei might have an anti-insulin resistance effect on insulin-resistance HepG2 cells in vitro and could be a promising candidate with anti-hyperglycemic properties for diabetes treatments.

Ethanol Extract of Eryngium Foetidum Leaves Induces Mitochondrial Associated Apoptosis via ROS Generation in Human Gastric Cancer Cells.

Background: Eryngium foetidum has long been used as a food ingredient and folk medicine in tropical regions. The anticancer activity of EF extract and the mechanisms remains unclear. Herein, we prepared four solvent extracts of EF leaves, detected the cytotoxic effects, and explored the potential mechanism by which these extracts induce cell death. Methods: The anticancer activity of the EF extracts was measured by MTT, CCK-8 and BrdU assays. The cell cycle was evaluated by flow cytometry and western blot. Apoptotic events were investigated with Hoechst, Annexin V/PI assays and western blot. The mitochondrial membrane potential was monitored using JC-1 staining, and ROS production was assessed with immunofluorescence. Results: The ethanol extract of EF leaves exhibited the strongest cytotoxic effect against SGC-7901 cells. The EFE extract significantly inhibited the SGC-7901 cells viability, arrested the cell cycle, increased the numbers of apoptotic cells, caused the loss of MMP, increased the Bax/Bcl-2 ratio, and led to cytochrome c release, and triggered ROS production. Conclusion: Our findings demonstrated for the first time that EFE extract induces mitochondrial associated apoptosis via ROS generation in SGC-7901 cells. Thus, EFE extract could be identified as a potential edible phytotherapy for the treatment of human gastric cancer.

Floral traits and reproductive success variation among inflorescence orders in Eryngium maritimum.

Eryngium maritimum L. is considered a key species in dunes of the western Mediterranean. Although Northern European populations are endangered, the Mediterranean populations show a good conservation status, despite a lack of studies of the reproductive requirements and strategies of this species. This study aims to analyse the reproductive biology of a population of E. maritimum in the western Mediterranean (Balearic Islands, Spain), specifically focusing on differences in morphometric and reproductive traits among inflorescence orders. Capitulum size, number of flowers, fruit set and seed set were analysed to evaluate the morphological and reproductive differences among inflorescence orders. The association between subterranean development and investment in reproductive structures was also studied along the dune location. Breeding system was analysed, evaluating self-pollination capacity at capitulum level and the pollen/ovary ratio. Finally, floral visitors were assessed. Morphometric and reproductive traits showed a decline in size and production from early to subsequent inflorescence orders. A correlation between width of stem base (caudex) and number of reproductive structures was observed, whereas seashore distance did not affect the reproductive development. Flowering occurred during summer, from May until late July, while fructification took place from the middle of August until the last days of September. Pollinator exclusion treatments induced a fruit and seed set decline. Twenty-one species of floral visitor were found. First and second inflorescence orders contributed to the generation of seeds, displaying high fruit and seed production, while other inflorescence orders seemed to attract pollinators. Reproductive and non-reproductive development could depend on the capacity to reallocate resources from underground to aerial structures, which seemed to be superior in samples with highly developed caudices. In the studied population, inner areas closest to the seashore showed a delay in anthesis, which could be related to microclimate differences along the coast-to-inland gradient. Pollination is strongly entomophilous and showed low levels of self-compatibility at capitulum level. Considering these results, for habitat conservation the reported microclimate diversity should be taken into consideration, with a focus on conservation of the pollinator populations.

Inhibition of enzymatic activities of Bothrops asper snake venom and docking analysis of compounds from plants used in Central America to treat snakebite envenoming.

ETHNOPHARMACOLOGICAL RELEVANCE: Snakebite envenoming is a public health problem of high impact in Central America. Bothrops asper, known as barba amarilla, terciopelo, and equis, is the snake species responsible for most snakebites in Central America. In this region, there is a long-standing tradition on the use of plants in the management of snakebites, especially in indigenous communities. Ethnomedical use of Eryngium foetidum L., Neurolaena lobata (L.) Cass. and Pimenta dioica (L.) Merr. to treat snakebite envenoming has been reported in Belice, Guatemala, Nicaragua, and Costa Rica. Extracts of the leaves of these plants have shown anti-venom activities in in vitro assays in previous studies. AIM OF THE STUDY: To assess the ability of organic fractions from these three plants to inhibit enzymatic activities associated with toxicity of the venom of B. asper, and to study, by docking analysis, the interaction of metalloproteinase and phospholipases A2 (PLA2) from B. asper venom with secondary metabolites previously described in these plants. MATERIALS AND METHODS: Organic fractions were obtained from these three plant species and their ability to neutralize proteolytic, PLA2 and in vitro coagulant activities of B. asper venom was assessed. A phytochemical analysis was carried out in these fractions. The interaction of secondary metabolites previously described in these plants with three toxins from B. asper venom (a metalloproteinase, a PLA2 and a PLA2 homologue) was investigated by docking analysis. RESULTS: The inhibitory activity of plants was mainly concentrated in their polar fractions. Acetonic fraction from P. dioica was the most active against PLA2 activity, while the acetonic fraction of E. foetidum completely inhibited the proteolytic activity of the venom. Coagulant activity was partially inhibited only by the acetone and ethyl acetate fractions of P. dioica. Phytochemical analysis of the most bioactive fractions identified flavonoids, saponins, essential oils, coumarins, alkaloids, tannins and sesquiterpene lactones. Docking analysis revealed high affinity interactions of several secondary metabolites of these plants with residues in the vicinity of the catalytic site of these enzymes and, in the case of PLA2 homologue myotoxin II, in the hydrophobic channel. CONCLUSIONS: Various fractions from these plants have inhibitory activity against enzymatic actions of B. asper venom which are directly associated with toxicological effects. Docking analysis showed structural evidence of the interaction of secondary metabolites with three toxins. These observations provide support to the potential of these plants to inhibit relevant toxic components of this snake venom.

Morpho-anatomical characterization of Eryngium yuccifolium ('Rattlesnake-master').

This paper provides detailed morpho-anatomical and histochemical characteristics of the leaves, peduncles, stems, and roots of Eryngium yuccifolium Michx. (Apiaceae) by bright-field light and scanning electron microscopy. Noteworthy morpho-anatomical features include the monocot-like leaves with parallel venation and distant marginal bristles, amphistomatic leaves with paracytic or anisocytic stomata and heterogenous isobilateral mesophyll, the presence of epicuticular wax crystalloids on both leaf surfaces, calcium oxalate druses and secretory canals in all the studied plant parts, presence of hypodermis, stem and peduncle with longitudinal ribs containing collenchyma, presence of aerenchyma in the peduncle and root, and absence of sclerenchyma in all the plant parts analyzed. These characteristics would aid in the taxonomy, species identification as well as quality control of the botanical.

Eryngium billardieri Extract and Fractions Induce Apoptosis in Cancerous Cells.

BACKGROUND: Eryngium is a genus flowering plant in the Umbelliferae family, having pharmacological properties, such as anti-inflammatory and anti-diabetic. Given the nature of melanoma and breast cancers in recent years and the fact that the anti-cancer properties of Eryngium billardieri on mentioned cell lines have not been studied, the present study was conducted to explore these properties. OBJECTIVE: The mechanisms of cytotoxicity and apoptosis of aerial parts of various extracts and fractions of E. billardieri on cancerous cells and normal cells were investigated. METHODS: Samples were collected from natural habitats, dried and then extracted by Soxhlet apparatus with solvents of n-Hex, DCM and methanol, respectively. The cytotoxic effects of the extracts were investigated by the MTT method on MCF7, B16 and HFF-2 classes for 24 and 48 hours. Flow cytometry was also used to investigate the mechanism of cytotoxicity, and it has been confirmed by Real-time PCR of p53 and Bax genes as to which comprise the apoptosis regulatory proteins. Meanwhile, volatile compounds of extracts were identified by the GC-MS method. RESULTS: The obtained data showed that the n-Hex extract of E. billardieri on B-16 and MCF7 cell lines and dichromethane extract on MCF7 cell line had the most significant cytotoxic effect compared to DMSO control (p-value <0.001). Our finding showed that the mechanism of n-Hex extract with 80% and 100% vlc fractions on B16 induced apoptotis compared to HFF-2 control cells; moreover, n-Hex extract and 80% vlc fraction on MCF7 were apoptotic. The major compounds of n-Hex, DCM, and 80% and 100% fractions of n-Hex extract obtained from GC-MS were non-terpenoid. CONCLUSION: Non-terpenoid compounds of E. billardieri can be responsible for exhibiting the most cytotoxic effects on MCF7 and B16 cell lines with apoptotic mechanism, and n-Hex extract was found to have the most significant inhibitory effect on cancerous cells compared to the HFF-2 control cells by employing the mechanism of apoptosis.

Novel Gold Nanoparticles: Green Synthesis with Eryngium thyrsoideum Boiss Extract, Characterization, and In Vivo Investigations on Inflammatory Gene Expression and Biochemical Parameters in Type 2 Diabetic Rats.

In this work, we synthesized and reported gold nanoparticles (Au NPs) with Eryngium thyrsoideum Boiss plant extract for first time. The plant extract has important effect as reducing and stabilizing agent for preparation of Au nanoparticles. The synthesized gold nanoparticles were characterized with FT-IR, UV-vis, XRD, SEM, and TEM analyses. All analyses confirmed successful synthesis of gold nanoparticles with high purity. The antidiabetic activity of synthesized Au NPs was investigated on type 2 diabetic rats by studying their influences on serum biochemical parameters and inflammatory markers. Obtained results revealed that hepatic enzymes, TNF-α, and interleukin-6 of diabetic rats receiving gold nanoparticles decreased in compare with healthy control rats. As inflammatory markers are main reasons for hyperglycemic-induced insulin resistance in diabetes, Au NPs have a possible option for management of the diabetes-related complication via their potent anti-inflammatory and hypoglycemic effects.

Synergistic Growth Inhibitory Effects of Eryngium kotschyi Extracts with Conventional Cytotoxic Agents: Cisplatin and Doxorubicin against Human Endometrium Cancer Cells.

BACKGROUND: Endometrial cancer is one of the most common types of cancer. For this reason, various studies have been carried out on its treatment and the effects of natural products on this disease. OBJECTIVES: This study aimed to examine the growth inhibitory effects of Eryngium kotschyi Boiss. ethyl acetate [EKE] and butanol [EKB] obtained from the main methanol [EKM] extract from the aerial parts on human endometrium carcinoma [RL95-2] cells and their synergistic effect with cisplatin or doxorubicin. METHODS: RL95-2 cells were treated with E. kotschyi extracts either alone or in combination with cisplatin or doxorubicin. The effects on cell growth were determined using the MTT assay and real-time cell analysis xCELLigence. RESULTS: The extracts demonstrated growth inhibitory activity, with a certain degree of selectivity against the RL95-2 cell line. Synergistic effects of EKE/cisplatin or doxorubicin at different concentration levels were demonstrated in RL95-2 cells. In some instances, the EKE/doxorubicin combinations resulted in antagonistic effects. The reduction level of cell viability was different and specific to each combination for the RL95-2 cell line. CONCLUSION: The growth inhibitory activity of cisplatin or doxorubicin, as a single agent, may be modified by combinations of the extracts and be synergistically enhanced in some cases. A significant synergistic effect of EKE on the RL95-2 cell line with cisplatin and doxorubicin was observed. This cytotoxic effect can be investigated in terms of molecular mechanisms. This study is the first of its kind in the literature. The mechanisms involved in this interaction between chemotherapeutic drugs and plant extracts remain unclear and should be further evaluated.

Cytotoxic evaluation and LC-MS/MS analysis of aerial parts of Eryngium kotschyi Boiss. grown in Turkey

Abstract Increasing biological activity and phytochemical investigations on Eryngium species showed its potential as pharmaceutical approach. Eryngium kotschyi Boiss. is one of the species of Eryngium genus and is endemic to Turkey. It is known that this plant is traditionally used in the South-western part of Turkey for the treatment of various diseases. This study focuses on cytotoxic activities of methanol extract and ethyl acetate, n-butanol and water sub-extracts from E. kotschyi in A549, COLO 205 and MDA-MB-231 cell lines by Sulforhodamin B assay and qualitative and quantitative determination of phytochemical constituents in active extract by LC-MS/MS. From the result of the study, it was seen that E. kotschyi ethyl acetate (EKE) sub-extract showed the strongest cytotoxic effect with the low IC50 values (50.00; 31.96 and 22.26 µg/mL in A549; COLO 205 and MDA-MB-231 cells at 48 h, respectively). Preliminary examination of the mass spectrums revealed the presence of 15 phytochemical compounds in active sub-extract and 7 of them was quantified. According to quantitative analyses the main compounds of EKE sub-extract were rosmarinic acid (485.603 µg/mgextract), chlorogenic acid (62.355 µg/mgextract) and caffeic acid (59.266 µg/mgextract). Moreover, this preliminary study on inhibitory activity of EKE sub-extract suggests further toxicologic investigations and detailed investigation on cytotoxic effect of various combinations of determined compounds

Effect of Elicitation with (+)-Usnic Acid on Accumulation of Phenolic Acids and Flavonoids in Agitated Microshoots of Eryngium alpinum L.

The present work was aimed at studying the potential of elicitation on the accumulation of phenolic compounds in in vitro shoot cultures of Eryngium alpinum L., a protected plant from the Apiaceae family. The study examined the influence of (+)-usnic acid on the biomass growth as well as on the biosynthesis of the desired flavonoids and phenolic acids in the cultured microshoots. The phenolic compound content was determined by HPLC-DAD. The flavonoid of the highest concentration was isoquercetin, and the phenolic acids of the highest amount were rosmarinic acid, caffeic acid and 3,4-dihydroxyphenylacetic acid, both in the non-elicited and elicited biomass. Isoquercetin accumulation was efficiently increased by a longer elicitation with a lower concentration of lichenic compound (107.17 ± 4.67 mg/100 g DW) or a shorter elicitation with a higher concentration of acid (127.54 ± 11.34 and 108.37 ± 12.1 mg/100 g DW). Rosmarinic acid production generally remained high in all elicited and non-elicited microshoots. The highest content of this acid was recorded at 24 h of elicitation with 3.125 µM usnic acid (512.69 ± 4.89 mg/100 g DW). The process of elicitation with (+)-usnic acid, a well-known lichenic compound with allelopathic nature, may therefore be an effective technique of enhancing phenolic compound accumulation in alpine eryngo microshoot biomass.

Contribution of fasting and postprandial glucose-lowering mechanisms to the acute hypoglycemic effect of traditionally used Eryngium cymosum F.Delaroche.

ETHNOPHARMACOLOGICAL RELEVANCE: Eryngium cymosum F. Delaroche was detected as a traditional remedy against type 2 diabetes consumed by patients of Tlanchinol in the state of Hidalgo, Mexico. AIM OF THE STUDY: Assessing the hypoglycemic effect and safety of the traditional extract of E. cymosum and relating it to key glucose-lowering mechanisms both in fasting and postprandial state. MATERIALS AND METHODS: The aqueous extract of E. cymosum was subjected to HPLC analysis to identify its main components. Hyperglycaemic STZ-NA Wistar rats were administered with the extract to evaluate its effect on blood glucose levels and a possible dose-dependence. Afterward, it was evaluated in both pyruvate and maltose tolerance tests in STZ-NA rats to characterize its effect on gluconeogenesis and carbohydrate breakdown, two of the main mechanisms responsible for fasting and postprandial hyperglycaemia in type 2 diabetes patients. In addition, the inhibitory capacity of the extract was evaluated on key enzymes involved in gluconeogenesis and a-glucosidases. Moreover, insulin concentrations were measured in normoglycemic rats in both conditions to establish a link between the hypoglycaemic effect of the extract with insulin release and functioning. RESULTS: Caffeic acid (1), chlorogenic acid (2), and rosmarinic acid (3) were identified as the main constituents of the aqueous extract of E. cymosum, which exerted a hypoglycaemic effect in hyperglycaemic STZ-NA rats. It has a significant antihyperglycemic effect in the pyruvate tolerance test, and it was able to reduce the postprandial hyperglycaemia in maltose tolerance tests significantly. Moreover, it effectively reduced the activity of both gluconeogenic enzymes reaching almost 100% of inhibition, while it presented a modest 32% inhibition of aglucosidases. On the other hand, the extract decreased insulin levels after its oral administration in healthy rats in both nutritional states, without affecting normoglycemia in normal curves and reducing the postprandial peak in glucose load curves. CONCLUSIONS: The traditional consumed form of aerial parts of E. cymosum is safe and regulated glucose levels both in fasting and in postprandial state.

Physiological performance and chemical compositions of the Eryngium foetidum L. (Apiaceae) essential oil cultivated with different fertilizer sources.

The present work proposed an evaluation of the physiological performance, yield and chemical profile of the essential oil obtained from leaves and roots of Eryngium foetidum L. (Apiaceae) cultivated using different fertilizer sources. The other gas exchange and chlorophyll a fluorescence parameter was not different, suggested that the photosystem II was not affected by the different fertilizer sources. Highest essential oil yield was obtained in leaf samples cultivated with the mineral fertilizer (0.18%) and control conditions (0.22%), while it was obtained by mineral fertilization in the case of root samples (0.22%). The principal component analysis - PCA and hierarchical cluster analysis - HCA showed two groups: the first was formed by roots (OrgR, MinR and ConR), characterized by the constituents 2,3,4-Trimethylbenzaldehyde, Muurola-4,10(14)-dien-1ß-ol, Isoshyobunone, para-Mint-1,5-dien-8-ol, 6-Canphenone, (E)-γ-Atlantone and (E)-ß-Farnesene; and the second group formed by leaves (OrgF, MinF, ConF), related to (2E)-2-Dodecenal, t-2-Tetradecenal, 1-Dodecanal, 1-Decanal, Tetradecanal, γ-Terpinene, Mesitylene and ρ-Cymene.