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Laurence Dinan - One of the best experts on this subject based on the ideXlab platform.
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small scale analysis of Phytoecdysteroids in seeds by hplc dad ms for the identification and quantification of specific analogues dereplication and chemotaxonomy
Phytochemical Analysis, 2020Co-Authors: Laurence Dinan, Christine Balducci, Louis Guibout, Rene LafontAbstract:INTRODUCTION: Phytoecdysteroids are analogues of arthropod steroids occurring in plants. They contribute to invertebrate deterrence. A wide diversity of ecdysteroids occurs in Phytoecdysteroid‐containing plant species, sometimes in high amounts. Ecdysteroids demonstrate potentially useful pharmaceutical actions in mammals. OBJECTIVES: Establish reversed‐phase high‐performance liquid chromatography with tandem mass spectrometry (RP‐HPLC‐MS/MS) and RP‐HPLC‐DAD‐MS (diode array detector mass spectrometry) methods for the separation, identification and quantification of ecdysteroids to screen for species containing significant amounts of 20‐hydroxyecdysone (20E) and other useful ecdysteroids. MATERIALS AND METHODS: Micro‐extracts of seed samples (ca. 30 mg) in 50% ethanol were subjected to RP‐SPE (solid‐phase extraction) purification prior to analysis by RP‐HPLC‐MS/MS and RP‐HPLC‐DAD‐MS. The method was initially applied to genera (Amaranthus, Centaurea, Lychnis, Ourisia, Serratula, Silene and Trollius) where high‐accumulating species had been previously encountered. Seeds of 160 randomly selected species, many of which have not previously been assessed, were then analysed. HPLC‐MS/MS with a short analysis time initially identifies ecdysteroid‐positive extracts and quantifies 20E. The positive extracts (20 ng 20E) are then analysed by HPLC‐MS/MS with a longer analysis time to identify and quantify 17 common Phytoecdysteroids and, finally, HPLC‐DAD‐MS (0.1–0.25 μg 20E) is used to obtain UV‐ and MS‐spectra to confirm identifications or as a basis for characterisation of partially identified or novel analogues. RESULTS: Lychnis coronaria, Silene fimbriata and Silene hookeri ecdysteroids are characterised for the first time and those of Cucubalus baccifer and Ipheion uniflorum are more extensively characterised. CONCLUSIONS: The procedure provides a rapid/sensitive method for screening small plant samples for the presence, quantification and identification of ecdysteroids. It permits ready dereplication of samples, identifying extracts containing large amounts or novel analogues.
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Distribution of Phytoecdysteroids in the Caryophyllaceae.
Phytochemistry, 2003Co-Authors: L. N. Zibareva, Rene Lafont, Tamara Savchenko, Pensri Whiting, Vladimir A. Volodin, Zyadilla Saatov, Laurence DinanAbstract:Abstract Certain genera within the Caryophyllaceae (especially Silene and Lychnis) have received a significant amount of attention with regard to the isolation and identification of ecdysteroids. However, the taxonomy of this family is difficult. Hence, the occurrence of Phytoecdysteroids in members of the Caryophyllaceae is presented, and combined with new data on ecdysteroid agonist (Phytoecdysteroid) and antagonist activities, in order to survey the distribution of Phytoecdysteroid-containing species within this large family, and to assess the utility of Phytoecdysteroids as chemotaxonomic markers. The new data presented (representing ca. 110 species) have been obtained by the application of sensitive biological/biochemical methods for the detection of ecdysteroid agonists and antagonists, using Drosophila melanogaster BII bioassay and ecdysteroid-specific immunoassays. In the antagonist version of the BII bioassay, only weak ecdysteroid antagonist activities were detected in a few of the extracts. From both new and previously available data, it was found that Phytoecdysteroids were present predominantly in the Genera Lychnis, Petrocoptis, Sagina and Silene. Comparison of ecdysteroid occurrence with a molecular phylogeny for the tribe Sileneae [Taxon 44 (1995) 525] revealed close association of ecdysteroid occurrence with certain groups of this tribe. In 14 species of Silene examined, there is a reasonable, but not absolute, relationship between the presence of ecdysteroids in the seeds and in other plant parts. Where ecdysteroids are present in the plant, highest concentrations are generally present in the roots.
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Phytoecdysteroids from the juice of Serratula coronata L. (Asteraceae).
Insect biochemistry and molecular biology, 2002Co-Authors: Victor N. Odinokov, Laurence Dinan, I. V. Galyautdinov, D. V. Nedopekin, Leonard M. Khalilov, A.s Shashkov, Vadim V. Kachala, Rene LafontAbstract:Abstract Seven Phytoecdysteroids have been isolated from Serratula coronata L.. One of them is a new Phytoecdysteroid, 3- epi -20-hydroxyecdysone. Two further ecdysteroids, 20-hydroxyecdysone 22-acetate and taxisterone, are isolated from this species for the first time in addition to the typical S. coronata ecdysteroids, 20-hydroxyecdysone, ecdysone, ajugasterone C and polypodine B. The juice squeezed from aerial parts of fresh plants of S. coronata was extracted with ethyl acetate. The ecdysteroids were isolated by a combination of chromatographic techniques (mainly HPLC) and identified by 1D and 2D 1 H and 13 C NMR experiments and mass-spectrometry. The biological activities of 3- epi -20-hydroxyecdysone ( EC 50 =1.6×10 −7 M), taxisterone ( EC 50 =9.5×10 −8 M) and ajugasterone C ( EC 50 =6.2×10 −8 M) have been determined in the Drosophila melanogaster B II bioassay for ecdysteroid agonist activity.
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Chemotaxonomic significance of ecdysteroid agonists and antagonists in the Ranunculaceae: Phytoecdysteroids in the genera Helleborus and Hepatica
Biochemical Systematics and Ecology, 2002Co-Authors: Laurence Dinan, Tamara Savchenko, Pensri WhitingAbstract:We present here a survey of ca. 100 species within 16 genera of the family Ranunculaceae for the presence of ecdysteroid agonist and antagonist activities in methanolic seed extracts. The levels of Phytoecdysteroids (agonists) have been quantified by radioimmunoassay and bioassay. A few samples possess weak antagonistic activity. Phytoecdysteroids are most prominently associated with the genus Helleborus. In this genus, species fall into two distinct classes: those with low or undetectable ecdysteroid levels and those with high ecdysteroid levels. The relationship between ecdysteroid levels and the biology of the plants in this genus is discussed. Additionally, the extract of Hepatica triloba Chaix seeds contains a significant level of Phytoecdysteroids. Several other species contain low levels of Phytoecdysteroids, as detected by radioimmunoassay. Together with our previous data on the genera Anemone and Pulsatilla, this survey allows us to present an overview of the distribution of ecdysteroids in this family.
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Phytoecdysteroid profiles in seeds of Sida spp. (Malvaceae).
Phytochemical analysis : PCA, 2001Co-Authors: Laurence Dinan, Pauline Bourne, Pensri WhitingAbstract:Procedures are presented for the assessment of the Phytoecdysteroid profiles in small plant samples (ca. 25 mg), using seeds of Sida spp. as an example. The procedures are suitable for the analysis of minute or valuable samples and provide copious information for chemotaxonomic purposes. Methanolic extracts of the plant material, after partioning against hexane, were separated by reversed-phase gradient HPLC monitored by PAD, RIA and bioassay. Aliquots of the fractions were also treated with Helix pomatia hydrolases, followed by RIA and bioassay, in order to assess the presence of hydrolysable ecdysteroid conjugates. Further information could also be obtained by separation of samples using normal-phase gradient HPLC. Among 11 species of Sida examined, seed extracts of S. acuta (=S.carpinifolia) and S. rhombifolia were found to contain significant amounts of ecdysteroids, seed extracts of S. filicaulis contained only moderate levels, whilst the remaining species showed no detectable levels of ecdysteroids. The ecdysteroid profiles of the extracts of the three positive species were significantly different, demonstrating that Phytoecdysteroids have chemotaxonomic value in this genus. Copyright © 2001 John Wiley & Sons, Ltd.
Rene Lafont - One of the best experts on this subject based on the ideXlab platform.
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small scale analysis of Phytoecdysteroids in seeds by hplc dad ms for the identification and quantification of specific analogues dereplication and chemotaxonomy
Phytochemical Analysis, 2020Co-Authors: Laurence Dinan, Christine Balducci, Louis Guibout, Rene LafontAbstract:INTRODUCTION: Phytoecdysteroids are analogues of arthropod steroids occurring in plants. They contribute to invertebrate deterrence. A wide diversity of ecdysteroids occurs in Phytoecdysteroid‐containing plant species, sometimes in high amounts. Ecdysteroids demonstrate potentially useful pharmaceutical actions in mammals. OBJECTIVES: Establish reversed‐phase high‐performance liquid chromatography with tandem mass spectrometry (RP‐HPLC‐MS/MS) and RP‐HPLC‐DAD‐MS (diode array detector mass spectrometry) methods for the separation, identification and quantification of ecdysteroids to screen for species containing significant amounts of 20‐hydroxyecdysone (20E) and other useful ecdysteroids. MATERIALS AND METHODS: Micro‐extracts of seed samples (ca. 30 mg) in 50% ethanol were subjected to RP‐SPE (solid‐phase extraction) purification prior to analysis by RP‐HPLC‐MS/MS and RP‐HPLC‐DAD‐MS. The method was initially applied to genera (Amaranthus, Centaurea, Lychnis, Ourisia, Serratula, Silene and Trollius) where high‐accumulating species had been previously encountered. Seeds of 160 randomly selected species, many of which have not previously been assessed, were then analysed. HPLC‐MS/MS with a short analysis time initially identifies ecdysteroid‐positive extracts and quantifies 20E. The positive extracts (20 ng 20E) are then analysed by HPLC‐MS/MS with a longer analysis time to identify and quantify 17 common Phytoecdysteroids and, finally, HPLC‐DAD‐MS (0.1–0.25 μg 20E) is used to obtain UV‐ and MS‐spectra to confirm identifications or as a basis for characterisation of partially identified or novel analogues. RESULTS: Lychnis coronaria, Silene fimbriata and Silene hookeri ecdysteroids are characterised for the first time and those of Cucubalus baccifer and Ipheion uniflorum are more extensively characterised. CONCLUSIONS: The procedure provides a rapid/sensitive method for screening small plant samples for the presence, quantification and identification of ecdysteroids. It permits ready dereplication of samples, identifying extracts containing large amounts or novel analogues.
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E- and Z-Isomers of New Phytoecdysteroid Conjugates from French Polynesian Microsorum membranifolium (Polypodiaceae) Fronds
Molecules, 2012Co-Authors: Jean-pierre Girault, Phila Raharivelomanana, Rene LafontAbstract:Phytochemical investigation of the fronds of Microsorum membranifolium resulted in the isolation of a new Phytoecdysteroid, E-2-deoxy-20-hydroxyecdysone 3-[4-(1-β-D-glucopyranosyl)]-caffeate (1), together with two known Phytoecdysteroids, E-2-deoxy-20-hydroxyecdysone 3-[4-(1-β-D-glucopyranosyl)]-ferulate (2), E-2-deoxyecdysone 3-[4-(1-β-D-glucopyranosyl)]-ferulate (3). Their respective Z-isomers 4-6 were also observed and identified for the first time. The new structures were elucidated on the basis of extensive spectroscopic data analysis (1D, 2D-NMR and HR-MS techniques).
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dietary effects of four Phytoecdysteroids on growth and development of the indian meal moth plodia interpunctella
Journal of Insect Science, 2010Co-Authors: Kacem Rharrabe, Fouad Sayah, Rene LafontAbstract:Using pure Phytoecdysteroids isolated from Ajuga iva (L.) Schreber (Lamiales: Lamiaceae) and Silene nutans L. (Caryophyllales: Caryophyllaceae), plants known for their high ecdysteroid content, a study was carried out on the effects of ingestion of four different Phytoecdysteroids (20-hydroxyecdysone, polypodine B, ponasterone A and makisterone A) on the growth and development of the Indian meal moth, Plodia interpunctella Hubner (Lepidoptera: Pyralidae) larvae when added at a concentration of 200 ppm in their diet. The experiments clearly showed the susceptibility of P. interpunctella to Phytoecdysteroid ingestion. The toxicity of Phytoecdysteroids manifested itself by a decrease in larval weight, induction of cannibalism and an increase of mortality, together with disruption of development. The severity of the Phytoecdysteroid effect on P. interpunctella depended on the structure of the molecule. The results demonstrate that the minimal structural differences existing between these four Phytoecdysteroids significantly affected their toxicity toward P. interpunctella. Makisterone A was the most toxic of the four compounds towards P. interpunctella larvae. In conclusion, Phytoecdysteroids ingestion evokes disruptive growth effects on P. interpunctella. This work supports a role for Phytoecdysteroids in plant defence against phytophagous insects.
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Distribution of Phytoecdysteroids in the Caryophyllaceae.
Phytochemistry, 2003Co-Authors: L. N. Zibareva, Rene Lafont, Tamara Savchenko, Pensri Whiting, Vladimir A. Volodin, Zyadilla Saatov, Laurence DinanAbstract:Abstract Certain genera within the Caryophyllaceae (especially Silene and Lychnis) have received a significant amount of attention with regard to the isolation and identification of ecdysteroids. However, the taxonomy of this family is difficult. Hence, the occurrence of Phytoecdysteroids in members of the Caryophyllaceae is presented, and combined with new data on ecdysteroid agonist (Phytoecdysteroid) and antagonist activities, in order to survey the distribution of Phytoecdysteroid-containing species within this large family, and to assess the utility of Phytoecdysteroids as chemotaxonomic markers. The new data presented (representing ca. 110 species) have been obtained by the application of sensitive biological/biochemical methods for the detection of ecdysteroid agonists and antagonists, using Drosophila melanogaster BII bioassay and ecdysteroid-specific immunoassays. In the antagonist version of the BII bioassay, only weak ecdysteroid antagonist activities were detected in a few of the extracts. From both new and previously available data, it was found that Phytoecdysteroids were present predominantly in the Genera Lychnis, Petrocoptis, Sagina and Silene. Comparison of ecdysteroid occurrence with a molecular phylogeny for the tribe Sileneae [Taxon 44 (1995) 525] revealed close association of ecdysteroid occurrence with certain groups of this tribe. In 14 species of Silene examined, there is a reasonable, but not absolute, relationship between the presence of ecdysteroids in the seeds and in other plant parts. Where ecdysteroids are present in the plant, highest concentrations are generally present in the roots.
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Phytoecdysteroids from the juice of Serratula coronata L. (Asteraceae).
Insect biochemistry and molecular biology, 2002Co-Authors: Victor N. Odinokov, Laurence Dinan, I. V. Galyautdinov, D. V. Nedopekin, Leonard M. Khalilov, A.s Shashkov, Vadim V. Kachala, Rene LafontAbstract:Abstract Seven Phytoecdysteroids have been isolated from Serratula coronata L.. One of them is a new Phytoecdysteroid, 3- epi -20-hydroxyecdysone. Two further ecdysteroids, 20-hydroxyecdysone 22-acetate and taxisterone, are isolated from this species for the first time in addition to the typical S. coronata ecdysteroids, 20-hydroxyecdysone, ecdysone, ajugasterone C and polypodine B. The juice squeezed from aerial parts of fresh plants of S. coronata was extracted with ethyl acetate. The ecdysteroids were isolated by a combination of chromatographic techniques (mainly HPLC) and identified by 1D and 2D 1 H and 13 C NMR experiments and mass-spectrometry. The biological activities of 3- epi -20-hydroxyecdysone ( EC 50 =1.6×10 −7 M), taxisterone ( EC 50 =9.5×10 −8 M) and ajugasterone C ( EC 50 =6.2×10 −8 M) have been determined in the Drosophila melanogaster B II bioassay for ecdysteroid agonist activity.
Mary Ann Lila - One of the best experts on this subject based on the ideXlab platform.
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Phytoecdysteroids Do Not Have Anabolic Effects in Skeletal Muscle in Sedentary Aging Mice.
International journal of environmental research and public health, 2021Co-Authors: Marcus M. Lawrence, Mary Ann Lila, Mary H. Grace, Kevin A. Zwetsloot, Susan T. Arthur, Chase A. Sherman, Joshua R. Huot, Vladimir Badmaev, David C. Nieman, R. Andrew ShanelyAbstract:Skeletal muscle mass and strength are lost with aging. Phytoecdysteroids, in particular 20-hydroxyecdysone (20E), increase protein synthesis in C2C12 skeletal muscle cells and muscle strength in young rats. The objective of this study was to determine whether an extract from Ajuga turkestanica (ATE), enriched in Phytoecdysteroids, and 20E affect skeletal muscle mass and fiber size, fiber type, activation of the PI3K–Akt signaling pathway, and the mRNA levels of MAFbx, MuRF-1, and myostatin in sedentary aging mice. Aging male C57BL/6 mice (20 months old) received ATE, 20E, or vehicle (CT) once per day for 28 days or a single acute dose. Treatment did not alter body, muscle, or organ mass; fiber cross-sectional area; or fiber type in the triceps brachii or plantaris muscles. Likewise, protein synthesis signaling markers (i.e., phosphorylation of AktSer473 and p70S6kThr389) measured after either 28 days or acutely were unchanged. Neither ATE nor 20E treatment for 28 days affected the mRNA levels of MAFbx, MuRF-1, and myostatin. In conclusion, these data indicate that Phytoecdysteroid treatment does not alter muscle mass or fiber type, nor does it activate protein synthesis signaling in the skeletal muscle of sedentary aging mice.
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Quinoa seeds leach Phytoecdysteroids and other compounds with anti-diabetic properties.
Food chemistry, 2014Co-Authors: Brittany L. Graf, Mary Ann Lila, Mary H. Grace, Alexander Poulev, Peter Kuhn, Ilya RaskinAbstract:Quinoa (Chenopodium quinoa Willd.) contains high levels of biologically active Phytoecdysteroids, which have been implicated in plant defense from insects, and have shown a range of beneficial pharmacological effects in mammals. We demonstrated that the most prevalent Phytoecdysteroid, 20-hydroxyecdysone (20HE), was secreted (leached) from intact quinoa seeds into water during the initial stages of seed germination. Leaching efficiency was optimized by ethanol concentration (70% ethanol), temperature (80°C), time (4h), and solvent ratio (5 ml/g seed). When compared to extraction of macerated seeds, the leaching procedure released essentially all the 20HE available in the seeds (491 μg/g seed). The optimized quinoa leachate (QL), containing 0.86% 20HE, 1.00% total Phytoecdysteroids, 2.59% flavonoid glycosides, 11.9% oil, and 20.4% protein, significantly lowered fasting blood glucose in obese, hyperglycemic mice. Leaching effectively releases and concentrates bioactive phytochemicals from quinoa seeds, providing an efficient means to produce a food-grade mixture that may be useful for anti-diabetic applications.
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variation in Phytoecdysteroid accumulation in seeds and shoots of spinacia oleracea l accessions
Hortscience, 2010Co-Authors: Diana M. Cheng, Gad G. Yousef, Mary Ann LilaAbstract:Spinach (Spinacea oleracea L.) is a valuable agricultural crop that accumulates Phytoecdysteroids, polyhydroxylatedtriterpenoids,whichmayplay a role in plant defense and have purported health benefits for human consumers. In this study, Phytoecdysteroid accumulation was measured in seeds and shoots of 15 spinach accessions to determine whether Phytoecdysteroid levels vary between spinach varieties and whether seed content could reliably predict relative levels in the edible foliage. Additionally, phytosterols, precursors to Phytoecdysteroids, were examined to determine potential points of reg- ulation of spinach Phytoecdysteroid biosynthesis. Significant variations in phytoecdyste- roidlevelsbetweenaccessionswereobserved(P <0.05),suggestingthepotentialforgenetic manipulation through traditional breeding or genetic engineering to increase phytoecdys- teroidlevelsinspinach.However,resultssuggestthatestimationofPhytoecdysteroidlevels inshootsmaynotbeachievedbymeasuringlevelsintheseeds.LevelsofPhytoecdysteroids in spinach ranged from 19.9 to 44.1 mg per shoot, 0.7 to 1.2 mgmg -1 dry mass shoot, 3.2 to 9.6 mg per seed, and 0.5 to 1.1 mgmg -1 seed. Several phytosterols connected to the Phytoecdysteroid biosynthetic pathway were identified by gas chromatography-mass spectroscopy, predominantlyspinasterol, 5-dihydroergosterol, and22-dihydrospinasterol, which comprised 79.8%, 6.3%, and 4.6% of the total phytosterol content, respectively. Detection of the phytosterols cycloartenol and lanosterol in spinach suggests that spinach mayalsohavedualbiosyntheticpathwaystophytosterolsthatcontributetotheproduction of Phytoecdysteroids.
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Variation in Phytoecdysteroid accumulation in seeds and shoots of Spinacia oleracea L. accessions
HortScience, 2010Co-Authors: Diana M. Cheng, Gad G. Yousef, Mary Ann LilaAbstract:Spinach (Spinacea oleracea L.) is a valuable agricultural crop that accumulates Phytoecdysteroids, polyhydroxylatedtriterpenoids,whichmayplay a role in plant defense and have purported health benefits for human consumers. In this study, Phytoecdysteroid accumulation was measured in seeds and shoots of 15 spinach accessions to determine whether Phytoecdysteroid levels vary between spinach varieties and whether seed content could reliably predict relative levels in the edible foliage. Additionally, phytosterols, precursors to Phytoecdysteroids, were examined to determine potential points of reg- ulation of spinach Phytoecdysteroid biosynthesis. Significant variations in phytoecdyste- roidlevelsbetweenaccessionswereobserved(P
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Phytoecdysteroids increase protein synthesis in skeletal muscle cells.
Journal of agricultural and food chemistry, 2008Co-Authors: Jonathan Gorelick-feldman, Diana M. Cheng, Mary Ann Lila, Alexander Poulev, David B. Maclean, Nebojša Ilić, Ilya RaskinAbstract:Phytoecdysteroids, which are structurally similar or identical to insect molting hormones, produce a range of effects in mammals, including increasing growth and physical performance. To study the mechanism of action of Phytoecdysteroids in mammalian tissue, an in vitro cellular assay of protein synthesis was developed. In C2C12 murine myotubes and human primary myotubes, Phytoecdysteroids increased protein synthesis by up to 20%. In vivo, ecdysteroids increased rat grip strength. Ecdysteroid-containing plant extracts produced similar results. The effect was inhibited by a phosphoinositide kinase-3 inhibitor, which suggests a PI3K-mediated mechanism.
Ilya Raskin - One of the best experts on this subject based on the ideXlab platform.
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Quinoa seeds leach Phytoecdysteroids and other compounds with anti-diabetic properties.
Food chemistry, 2014Co-Authors: Brittany L. Graf, Mary Ann Lila, Mary H. Grace, Alexander Poulev, Peter Kuhn, Ilya RaskinAbstract:Quinoa (Chenopodium quinoa Willd.) contains high levels of biologically active Phytoecdysteroids, which have been implicated in plant defense from insects, and have shown a range of beneficial pharmacological effects in mammals. We demonstrated that the most prevalent Phytoecdysteroid, 20-hydroxyecdysone (20HE), was secreted (leached) from intact quinoa seeds into water during the initial stages of seed germination. Leaching efficiency was optimized by ethanol concentration (70% ethanol), temperature (80°C), time (4h), and solvent ratio (5 ml/g seed). When compared to extraction of macerated seeds, the leaching procedure released essentially all the 20HE available in the seeds (491 μg/g seed). The optimized quinoa leachate (QL), containing 0.86% 20HE, 1.00% total Phytoecdysteroids, 2.59% flavonoid glycosides, 11.9% oil, and 20.4% protein, significantly lowered fasting blood glucose in obese, hyperglycemic mice. Leaching effectively releases and concentrates bioactive phytochemicals from quinoa seeds, providing an efficient means to produce a food-grade mixture that may be useful for anti-diabetic applications.
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ecdysteroids elicit a rapid ca2 flux leading to akt activation and increased protein synthesis in skeletal muscle cells
Steroids, 2010Co-Authors: Jonathan Gorelickfeldman, Wendie S Cohick, Ilya RaskinAbstract:Phytoecdysteroids, structurally similar to insect molting hormones, produce a range of effects in mammals, including increasing growth and physical performance. In skeletal muscle cells, Phytoecdysteroids increase protein synthesis. In this study we show that in a mouse skeletal muscle cell line, C(2)C(12), 20-hydroxyecdysone (20HE), a common Phytoecdysteroid in both insects and plants, elicited a rapid elevation in intracellular calcium, followed by sustained Akt activation and increased protein synthesis. The effect was inhibited by a G-protein coupled receptor (GPCR) inhibitor, a phospholipase C (PLC) inhibitor, and a phosphoinositide kinase-3 (PI3K) inhibitor.
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Phytoecdysteroids increase protein synthesis in skeletal muscle cells.
Journal of agricultural and food chemistry, 2008Co-Authors: Jonathan Gorelick-feldman, Diana M. Cheng, Mary Ann Lila, Alexander Poulev, David B. Maclean, Nebojša Ilić, Ilya RaskinAbstract:Phytoecdysteroids, which are structurally similar or identical to insect molting hormones, produce a range of effects in mammals, including increasing growth and physical performance. To study the mechanism of action of Phytoecdysteroids in mammalian tissue, an in vitro cellular assay of protein synthesis was developed. In C2C12 murine myotubes and human primary myotubes, Phytoecdysteroids increased protein synthesis by up to 20%. In vivo, ecdysteroids increased rat grip strength. Ecdysteroid-containing plant extracts produced similar results. The effect was inhibited by a phosphoinositide kinase-3 inhibitor, which suggests a PI3K-mediated mechanism.
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In vitro production of metabolism-enhancing Phytoecdysteroids from Ajuga turkestanica
Plant Cell Tissue and Organ Culture, 2008Co-Authors: Diana M. Cheng, Gad G. Yousef, Mary H. Grace, Randy B. Rogers, Jonathan Gorelick-feldman, Ilya Raskin, Mary Ann LilaAbstract:In order to develop a sustainable source of metabolism-enhancing Phytoecdysteroids, cell suspension and hairy root cultures were established from shoot cultures of wild-harvested Ajuga turkestanica, a medicinal plant indigenous to Uzbekistan. Precursors of Phytoecdysteroids (acetate, mevalonic acid cholesterol) or methyl jasmonate (an elicitor) were added to subculture media to increase Phytoecdysteroid accumulation. In cell suspension cultures, 20-hydroxyecdysone (20E) content increased 3- or 2-fold with the addition of 125 or 250 μM methyl jasmonate, respectively, compared to unelicited cultures. Precursor addition, however, did not provoke Phytoecdysteroid accumulation. In hairy root cultures, addition of sodium acetate, mevalonic acid, and methyl jasmonate, but not cholesterol, increased Phytoecdysteroid content compared to unelicited cultures. Hairy root cultures treated with 150 mg l−1 sodium acetate, or 15 or 150 mg l−1 mevalonic acid, increased 20E content approximately 2-fold to 19.9, 20.4 or 21.7 μg mg−1, respectively, compared to control (10.5 μg mg−1). Older hairy root cultures, extracted after the seventh subculture cycle, also showed increases in 20E content (24.8 μg mg−1), turkesterone (0.9 μg mg−1) and cyasterone (8.1 μg mg−1) compared to control cultures maintained for a shorter duration of four subculture cycles. Doses of 10 or 20 μg ml−1 hairy root extract increased protein synthesis by 25.7% or 31.1%, respectively, in a C2C12 mouse skeletal cell line. These results suggest that sustainable production of metabolically active Phytoecdysteroid can be achieved through hairy root culture systems.
Diana M. Cheng - One of the best experts on this subject based on the ideXlab platform.
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variation in Phytoecdysteroid accumulation in seeds and shoots of spinacia oleracea l accessions
Hortscience, 2010Co-Authors: Diana M. Cheng, Gad G. Yousef, Mary Ann LilaAbstract:Spinach (Spinacea oleracea L.) is a valuable agricultural crop that accumulates Phytoecdysteroids, polyhydroxylatedtriterpenoids,whichmayplay a role in plant defense and have purported health benefits for human consumers. In this study, Phytoecdysteroid accumulation was measured in seeds and shoots of 15 spinach accessions to determine whether Phytoecdysteroid levels vary between spinach varieties and whether seed content could reliably predict relative levels in the edible foliage. Additionally, phytosterols, precursors to Phytoecdysteroids, were examined to determine potential points of reg- ulation of spinach Phytoecdysteroid biosynthesis. Significant variations in phytoecdyste- roidlevelsbetweenaccessionswereobserved(P <0.05),suggestingthepotentialforgenetic manipulation through traditional breeding or genetic engineering to increase phytoecdys- teroidlevelsinspinach.However,resultssuggestthatestimationofPhytoecdysteroidlevels inshootsmaynotbeachievedbymeasuringlevelsintheseeds.LevelsofPhytoecdysteroids in spinach ranged from 19.9 to 44.1 mg per shoot, 0.7 to 1.2 mgmg -1 dry mass shoot, 3.2 to 9.6 mg per seed, and 0.5 to 1.1 mgmg -1 seed. Several phytosterols connected to the Phytoecdysteroid biosynthetic pathway were identified by gas chromatography-mass spectroscopy, predominantlyspinasterol, 5-dihydroergosterol, and22-dihydrospinasterol, which comprised 79.8%, 6.3%, and 4.6% of the total phytosterol content, respectively. Detection of the phytosterols cycloartenol and lanosterol in spinach suggests that spinach mayalsohavedualbiosyntheticpathwaystophytosterolsthatcontributetotheproduction of Phytoecdysteroids.
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Variation in Phytoecdysteroid accumulation in seeds and shoots of Spinacia oleracea L. accessions
HortScience, 2010Co-Authors: Diana M. Cheng, Gad G. Yousef, Mary Ann LilaAbstract:Spinach (Spinacea oleracea L.) is a valuable agricultural crop that accumulates Phytoecdysteroids, polyhydroxylatedtriterpenoids,whichmayplay a role in plant defense and have purported health benefits for human consumers. In this study, Phytoecdysteroid accumulation was measured in seeds and shoots of 15 spinach accessions to determine whether Phytoecdysteroid levels vary between spinach varieties and whether seed content could reliably predict relative levels in the edible foliage. Additionally, phytosterols, precursors to Phytoecdysteroids, were examined to determine potential points of reg- ulation of spinach Phytoecdysteroid biosynthesis. Significant variations in phytoecdyste- roidlevelsbetweenaccessionswereobserved(P
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Phytoecdysteroids increase protein synthesis in skeletal muscle cells.
Journal of agricultural and food chemistry, 2008Co-Authors: Jonathan Gorelick-feldman, Diana M. Cheng, Mary Ann Lila, Alexander Poulev, David B. Maclean, Nebojša Ilić, Ilya RaskinAbstract:Phytoecdysteroids, which are structurally similar or identical to insect molting hormones, produce a range of effects in mammals, including increasing growth and physical performance. To study the mechanism of action of Phytoecdysteroids in mammalian tissue, an in vitro cellular assay of protein synthesis was developed. In C2C12 murine myotubes and human primary myotubes, Phytoecdysteroids increased protein synthesis by up to 20%. In vivo, ecdysteroids increased rat grip strength. Ecdysteroid-containing plant extracts produced similar results. The effect was inhibited by a phosphoinositide kinase-3 inhibitor, which suggests a PI3K-mediated mechanism.
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In vitro production of metabolism-enhancing Phytoecdysteroids from Ajuga turkestanica
Plant Cell Tissue and Organ Culture, 2008Co-Authors: Diana M. Cheng, Gad G. Yousef, Mary H. Grace, Randy B. Rogers, Jonathan Gorelick-feldman, Ilya Raskin, Mary Ann LilaAbstract:In order to develop a sustainable source of metabolism-enhancing Phytoecdysteroids, cell suspension and hairy root cultures were established from shoot cultures of wild-harvested Ajuga turkestanica, a medicinal plant indigenous to Uzbekistan. Precursors of Phytoecdysteroids (acetate, mevalonic acid cholesterol) or methyl jasmonate (an elicitor) were added to subculture media to increase Phytoecdysteroid accumulation. In cell suspension cultures, 20-hydroxyecdysone (20E) content increased 3- or 2-fold with the addition of 125 or 250 μM methyl jasmonate, respectively, compared to unelicited cultures. Precursor addition, however, did not provoke Phytoecdysteroid accumulation. In hairy root cultures, addition of sodium acetate, mevalonic acid, and methyl jasmonate, but not cholesterol, increased Phytoecdysteroid content compared to unelicited cultures. Hairy root cultures treated with 150 mg l−1 sodium acetate, or 15 or 150 mg l−1 mevalonic acid, increased 20E content approximately 2-fold to 19.9, 20.4 or 21.7 μg mg−1, respectively, compared to control (10.5 μg mg−1). Older hairy root cultures, extracted after the seventh subculture cycle, also showed increases in 20E content (24.8 μg mg−1), turkesterone (0.9 μg mg−1) and cyasterone (8.1 μg mg−1) compared to control cultures maintained for a shorter duration of four subculture cycles. Doses of 10 or 20 μg ml−1 hairy root extract increased protein synthesis by 25.7% or 31.1%, respectively, in a C2C12 mouse skeletal cell line. These results suggest that sustainable production of metabolically active Phytoecdysteroid can be achieved through hairy root culture systems.