RESUMO
Glycogen synthase kinase 3ß (GSK3ß) is considered an important element of glycogen metabolism; however, it has many other regulatory roles. Changes in the GSK3ß signaling mechanism have been associated with various disorders, such as Alzheimer's disease (AD), type II diabetes, and cancer. Although the effects of GSK3ß inhibitors on reducing the pathological effects of AD have been described, an effective inhibitor has not yet been developed. Epibrassinolide (EBR), a brassinosteroid (BR), is structurally similar to mammalian steroid hormones. Our studies have shown that EBR has an inhibitory effect on GSK3ß in different cell lines. Roscovitine (ROSC), a cyclin-dependent kinase (CDK) inhibitor, has also been identified as a potential GSK3 inhibitor. Within the scope of this study, we propose that EBR and/or ROSC might have mechanistic action in AD models. To test this hypothesis, we used in vitro models and Caenorhabditis elegans (C. elegans) AD strains. Finally, EBR treatment successfully protected cells from apoptosis and increased the inhibitory phosphorylation of GSK3ß. In addition, EBR and/or ROSC treatment had a positive effect on the survival rates of C. elegans strains. More interestingly, the paralysis phenotype of the C. elegans AD model due to Aß42 toxicity was prevented by EBR and/or ROSC. Our findings suggest that EBR and ROSC administration have neuroprotective effects on both in vitro and C. elegans models via inhibitory GSK3ß phosphorylation at Ser9.
Assuntos
Brassinosteroides/farmacologia , Caenorhabditis elegans/crescimento & desenvolvimento , Glicogênio Sintase Quinase 3 beta/antagonistas & inibidores , Longevidade , Transtornos Motores/tratamento farmacológico , Roscovitina/farmacologia , Esteroides Heterocíclicos/farmacologia , Proteínas tau/metabolismo , Animais , Brassinosteroides/química , Caenorhabditis elegans/efeitos dos fármacos , Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/metabolismo , Quimioterapia Combinada , Glicogênio Sintase Quinase 3 beta/genética , Glicogênio Sintase Quinase 3 beta/metabolismo , Fármacos Neuroprotetores/farmacologia , Fosforilação , Reguladores de Crescimento de Plantas/química , Reguladores de Crescimento de Plantas/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Esteroides Heterocíclicos/química , Proteínas tau/genéticaRESUMO
Brassinosteroids are polyhydroxysteroids that are involved in different plants' biological functions, such as growth, development and resistance to biotic and external stresses. Because of its low abundance in plants, much effort has been dedicated to the synthesis and characterization of brassinosteroids analogs. Herein, we report the synthesis of brassinosteroid 24-nor-5ß-cholane type analogs with 23-benzoate function and 22,23-benzoate groups. The synthesis was accomplished with high reaction yields in a four-step synthesis route and using hyodeoxycholic acid as starting material. All synthesized analogs were tested using the rice lamina inclination test to assess their growth-promoting activity and compare it with those obtained for brassinolide, which was used as a positive control. The results indicate that the diasteroisomeric mixture of monobenzoylated derivatives exhibit the highest activity at the lowest tested concentrations (1 × 10-8 and 1 × 10-7 M), being even more active than brassinolide. Therefore, a simple synthetic procedure with high reaction yields that use a very accessible starting material provides brassinosteroid synthetic analogs with promising effects on plant growth. This exploratory study suggests that brassinosteroid analogs with similar chemical structures could be a good alternative to natural brassinosteroids.
Assuntos
Benzoatos/síntese química , Brassinosteroides/síntese química , Colanos/síntese química , Desenvolvimento Vegetal , Arabidopsis/crescimento & desenvolvimento , Benzoatos/química , Brassinosteroides/química , Colanos/química , Ácido Desoxicólico/síntese química , Ácido Desoxicólico/química , Estrutura Molecular , Oryza/química , Reguladores de Crescimento de Plantas , Esteroides Heterocíclicos/químicaRESUMO
The postharvest senescence accompanied by yellowing limited the shelf-life of broccoli. In this study, we developed a novel W/O/W double emulsion co-delivering brassinolide and cinnamon essential oil and applied it to broccoli for preservation. Results showed that double emulsion prepared by whey protein concentrate-high methoxyl pectin (1:3) exhibited best storage stability with largest particle size (581.30 nm), lowest PDI (0.23) and zeta potential (-40.31 mV). This double emulsion also exhibited highest encapsulation efficiency of brassinolide (92%) and cinnamon essential oil (88%). The broccoli coated with double emulsion maintained higher chlorophyll contents and activities of chlorophyllase and magnesium-dechelatase were reduced by 9% and 24%, respectively. The energy metabolic enzymes (SDH, CCO, H+-ATPase, Ca2+-ATPase) were also activated, inducing higher level of ATP and energy charge. These results demonstrated W/O/W double emulsion co-delivering brassinolide and cinnamon essential delayed the senescence of broccoli via regulating chlorophyll degradation and energy metabolism.
Assuntos
Brassica/metabolismo , Brassinosteroides/química , Clorofila/metabolismo , Emulsões/química , Metabolismo Energético , Óleos Voláteis/química , Esteroides Heterocíclicos/química , Adenosina Trifosfatases/química , Adenosina Trifosfatases/metabolismo , Trifosfato de Adenosina/metabolismo , Brassica/efeitos dos fármacos , Brassinosteroides/metabolismo , Brassinosteroides/farmacologia , Hidrolases de Éster Carboxílico/antagonistas & inibidores , Hidrolases de Éster Carboxílico/metabolismo , Cinnamomum zeylanicum/metabolismo , Emulsões/metabolismo , Metabolismo Energético/efeitos dos fármacos , Enzimas/química , Armazenamento de Alimentos/métodos , Óleos Voláteis/metabolismo , Óleos Voláteis/farmacologia , Tamanho da Partícula , Esteroides Heterocíclicos/metabolismo , Esteroides Heterocíclicos/farmacologia , ViscosidadeRESUMO
Brassinosteroids (BRs) are plant hormones of steroid nature, regulating various developmental and adaptive processes. The perception, transport, and signaling of BRs are actively studied nowadays via a wide range of biochemical and genetic tools. However, most of the knowledge about BRs intracellular localization and turnover relies on the visualization of the receptors or cellular compartments using dyes or fluorescent protein fusions. We have previously synthesized a conjugate of epibrassinolide with green fluorescent dye BODIPY (eBL-BODIPY). Here we present a detailed assessment of the compound bioactivity and its suitability as probe for in vivo visualization of BRs. We show that eBL-BODIPY rapidly penetrates epidermal cells of Arabidopsis thaliana roots and after long exposure causes physiological and transcriptomic responses similar to the natural hormone.
Assuntos
Compostos de Boro/química , Brassinosteroides/química , Corantes Fluorescentes/química , Esteroides Heterocíclicos/química , Arabidopsis/metabolismo , Brassinosteroides/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Raízes de Plantas/metabolismo , Transdução de SinaisRESUMO
A priority of modern agriculture is to use novel and environmentally friendly plant-growth promoter compounds to increase crop yields and avoid the indiscriminate use of synthetic fertilizers. Brassinosteroids are directly involved in the growth and development of plants and are considered attractive candidates to solve this problem. Obtaining these metabolites from their natural sources is expensive and cumbersome since they occur in extremely low concentrations in plants. For this reason, much effort has been dedicated in the last decades to synthesize brassinosteroids analogs. In this manuscript, we present the synthesis and characterization of seven steroidal carbamates starting from stigmasterol, ß-sitosterol, diosgenin and several oxygenated derivatives of it. The synthesis route for functionalization of diosgenin included epoxidation and epoxy opening reactions, reduction of carbonyl groups, selective oxidation of hydroxyl groups, among others. All the obtained compounds were characterized by 1H and 13C NMR, HRMS, and their melting points are also reported. Rice lamina inclination test performed at different concentrations established that all reported steroidal carbamates show plant-growth-promoting activity. A molecular docking study evaluated the affinity of the synthesized compounds towards the BRI1-BAK1 receptor from Arabidopsis thaliana and three of the docked compounds displayed a binding energy lower than brassinolide.
Assuntos
Arabidopsis/crescimento & desenvolvimento , Carbamatos , Simulação de Acoplamento Molecular , Oryza/crescimento & desenvolvimento , Reguladores de Crescimento de Plantas , Brassinosteroides/química , Carbamatos/síntese química , Carbamatos/química , Carbamatos/farmacologia , Reguladores de Crescimento de Plantas/síntese química , Reguladores de Crescimento de Plantas/química , Reguladores de Crescimento de Plantas/farmacologia , Esteroides Heterocíclicos/químicaRESUMO
The effects of individual epi-brassinolide (eBL) and NaCl, as well as their combination on contents of main phytochemicals and antioxidant capacity of Chinese kale sprouts were investigated. Our results showed that the application of 100â¯nM eBL decreased the contents of individual and total glucosinolates, while treatments of 160â¯mM NaCl both alone and combined with 100â¯nM eBL enhanced the glucosinolates accumulation by promoting the expression of genes involved in glucosinolate biosynthesis in Chinese kale sprouts and the combined treatment led to significantly higher content of most glucosinolate profiles. Moreover, it also elevated the contents of ascorbic acid and total carotenoids, whereas did not influence the total phenolics and antioxidant capacity. These findings indicated that the combined treatment of 100â¯nM eBL plus 160â¯mM NaCl could provide a new strategy to improve the main health promoting compounds in Chinese kale sprouts.
Assuntos
Brassica/química , Brassinosteroides/química , Compostos Fitoquímicos/química , Plântula/química , Cloreto de Sódio/química , Esteroides Heterocíclicos/química , Antioxidantes/química , Ácido Ascórbico/química , Carotenoides/química , Glucosinolatos/química , Fenóis/química , Cloreto de Sódio/farmacologiaRESUMO
The present study unravels origin of nitric oxide (NO) and the interaction between 24-Epibrassinolide (EBL) and nitrate reductase (NR) for NO production in Indian mustard (Brassica juncea L.) under salinity stress. Two independent experiments were performed to check whether (i) Nitrate reductase or Nitric oxide synthase takes part in the biosynthesis of endogenous NO and (ii) EBL has any regulatory effect on NR-dependent NO biosynthesis in the alleviation of salinity stress. Results revealed that NR-inhibitor tungstate significantly (P ≤ 0.05) decreased the NR activity and endogenous NO content, while NOS inhibitor l-NAME did not influence NO biosynthesis and plant growth. Under salinity stress, inhibition in NR activity decreased the activities of antioxidant enzymes, increased H2O2, MDA, protein carbonyl content and caused DNA damage, implying that antioxidant defense might be related to NO signal. EBL supplementation enhanced the NR activity but did not influence NOS activity, suggesting that NR was involved in endogenous NO production. EBL supplementation alleviated the inhibitory effects of salinity stress and improved the plant growth by enhancing nutrients, photosynthetic pigments, compatible osmolytes, and performance of AsA-GSH cycle. It also decreased the superoxide ion accumulation, leaf epidermal damages, cell death, DNA damage, and ABA content. Comet assay revealed significant (P ≤ 0.05) enhancement in tail length and olive tail moment, while flow cytometry did not showed any significant (P ≤ 0.05) changes in genome size and ploidy level under salinity stress. Moreover, EBL supplementation increased the G6PDH activity and S-nitrosothiol content which further boosted the antioxidant responses under salinity stress. Taken together, these results suggested that NO production in mustard occurred in NR-dependent manner and EBL in association with endogenous NO activates the antioxidant system to counter salinity stress.
Assuntos
Brassinosteroides/metabolismo , Mostardeira/química , Nitrato Redutase/metabolismo , Óxido Nítrico/biossíntese , Estresse Salino , Esteroides Heterocíclicos/metabolismo , Brassinosteroides/química , Inibidores Enzimáticos/farmacologia , Índia , Mostardeira/metabolismo , NG-Nitroarginina Metil Éster/farmacologia , Nitrato Redutase/química , Óxido Nítrico/química , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase/metabolismo , Folhas de Planta/química , Folhas de Planta/metabolismo , Esteroides Heterocíclicos/químicaRESUMO
Brassinosteroids (BRs) are essential phytohormones, which bind to the plant receptor, BRI1, to regulate various physiological processes. The molecular mechanism of the perception of BRs by the ectodomain of BRI1 remains not fully understood. It also remains elusive why a substantial difference in biological activity exists between the BRs. In this work, we study the binding mechanisms of the two most bioactive BRs, brassinolide (BLD) and castasterone (CAT), using molecular dynamics simulations. We report free-energy landscapes of the binding processes of both ligands, as well as detailed ligand binding pathways. Our results suggest that CAT has a lower binding affinity compared to BLD due to its inability to form hydrogen-bonding interactions with a tyrosine residue in the island domain of BRI1. We uncover a conserved nonproductive binding state for both BLD and CAT, which is more stable for CAT and may further contribute to the bioactivity difference. Finally, we validate past observations about the conformational restructuring and ordering of the island domain upon BLD binding. Overall, this study provides new insights into the fundamental mechanism of the perception of the two most bioactive BRs, which may create new avenues for genetic and agrochemical control of their signaling cascade.
Assuntos
Proteínas de Arabidopsis/metabolismo , Brassinosteroides/metabolismo , Colestanóis/metabolismo , Proteínas Quinases/metabolismo , Esteroides Heterocíclicos/metabolismo , Arabidopsis/química , Proteínas de Arabidopsis/química , Brassinosteroides/química , Colestanóis/química , Ligação de Hidrogênio , Ligantes , Modelos Químicos , Simulação de Dinâmica Molecular , Ligação Proteica , Conformação Proteica , Proteínas Quinases/química , Esteroides Heterocíclicos/química , Termodinâmica , Tirosina/químicaRESUMO
Lupinus angustifolius L. is a legume culture known as a source of valuable feed protein and the N2-fixator for improving soil fertility. However, its low ecological resistance does not allow for a stable yield of the crop. Earlier, we have shown that steroid phytohormone 24-epibrassinolide (EBR) increases the tolerance of lupine to chlorine ions by activating the protective proteins in ripening seeds (such as proteinase inhibitors that prevent protein breakdown) and lectins. Here we investigated the effect of EBR on the functional status of the N2-fixing system in root nodules, protein synthesis in ripening seeds and the resistance of lupine plants to various pathogens. It was found that EBR enhanced the nodulation process, N2-fixing activity of nitrogenase and the accumulation of poly-ß-hydroxybutirate in the bacteroides, increased the leghemoglobin content in the nodules as well as the metabolic activity of bacteroides. According to data on the inclusion of 14C-leucine in maturing seed proteins, EBR increased the accumulation of protein in them against the background of a short-term decrease in protein synthesis and its subsequent regeneration to the control level. Gradual inhibition of protein synthesis, characteristic of other legumes, was observed in control variants of lupine. EBR increased lupine resistance to phytopathogenic fungi of Colletotrichum genus and insects of Noctuidae and Scarabaeidae families. We concluded that a more complete implementation of the potential productivity and sustainability of lupine under the action of EBR was achieved due to the anabolic/anti-catabolic effect on the N2 fixation system in root nodules, as well as on protein synthesis in ripening seeds.
Assuntos
Antifúngicos/farmacologia , Brassinosteroides/farmacologia , Inseticidas/farmacologia , Lupinus/química , Reguladores de Crescimento de Plantas/farmacologia , Sementes/química , Esteroides Heterocíclicos/farmacologia , Animais , Antifúngicos/química , Antifúngicos/metabolismo , Brassinosteroides/química , Brassinosteroides/metabolismo , Besouros/efeitos dos fármacos , Colletotrichum/efeitos dos fármacos , Inseticidas/química , Inseticidas/metabolismo , Lupinus/metabolismo , Testes de Sensibilidade Microbiana , Reguladores de Crescimento de Plantas/química , Reguladores de Crescimento de Plantas/metabolismo , Sementes/metabolismo , Spodoptera/efeitos dos fármacos , Esteroides Heterocíclicos/química , Esteroides Heterocíclicos/metabolismoRESUMO
Nanoscale supramolecular systems have been increasingly gaining importance as drug release vehicles due to their ability to target tumor cells. In this work, we have developed a new class of nanoassemblies derived from the phytosterol 24-EpiBrassinolide (EpiB) for the development of nanocarriers for the anticancer drug Doxorubicin (DOX). EpiB is a biocompatible cholesterol mimic, and has inherent apoptotic properties toward cancer cells. Thus, by encapsulating DOX within a nanocarrier with innate anticancer ability we have developed a targeting system that can enhance the uptake and efficacy of DOX in tumor cells. The nanocarriers were formed by self-assembly of EpiB. The morphologies of assemblies formed were dependent upon the concentration of EpiB used. While at low concentrations, spherical nanoassemblies were formed, at higher concentration, lamellar aggregates with birefringence properties were observed. Our results indicated that the drug loaded nanocarriers showed diffusion controlled release of the drug, and demonstrated antiproliferative effects, cellular uptake and were apoptotic against HeLa cervical cancer cells. Furthermore, EpiB loaded DOX enhanced both apoptosis and uptake into the cell's nuclei. These supramolecular assemblies may have potential applications for enhancing efficacy of chemotherapeutic drugs through passive targeting.
Assuntos
Antibióticos Antineoplásicos/química , Doxorrubicina/química , Portadores de Fármacos/química , Nanoestruturas/química , Fitosteróis/química , Antibióticos Antineoplásicos/metabolismo , Antibióticos Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Brassinosteroides/química , Doxorrubicina/metabolismo , Doxorrubicina/farmacologia , Liberação Controlada de Fármacos , Células HeLa , Humanos , Microscopia Confocal , Espectroscopia de Infravermelho com Transformada de Fourier , Esteroides Heterocíclicos/químicaRESUMO
The progressive contamination of food products by mycotoxins such as zearalenone (ZEN) has prompted the search for specific substances that can act as protectors against an accumulation of these toxins. This paper discusses the effect of selenium ions and 24-epibrassinolide (EBR) as non-organic and organic compounds that preserve human lymphoblastic cells U-937 under ZEN stressogenic conditions. Based on measurements of cell viability and a DAPI test, concentrations of ZEN at 30 µmol/l, Se at 2.5 µmol/l and EBR at 0.005 µmol/l were selected. The addition of both protectors resulted in an increase in the viability of ZEN-treated cells by about 16%. This effect was connected with a decrease in lipid peroxidation (a decrease in the malonyldialdehyde content) and the generation of reactive oxygen species, which were determined by a cellular ROS/superoxide detection assay and the SOD activity. The Se protection was observed as the blocking of the all excess ROS, while the EBR action was mainly concentrated on something other than the superoxide radical itself. The experiments on the model lipid membranes that mimic the environment of U-937 cells confirmed the affect of ZEN on the structure and physicochemical properties of human membranes. Although the presence of both Se and EBR reduced the effect of ZEN by blocking its interaction with a membrane, the action of Se was more evident.
Assuntos
Brassinosteroides/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Micotoxinas/farmacologia , Selênio/farmacologia , Esteroides Heterocíclicos/farmacologia , Brassinosteroides/química , Membrana Celular , Fragmentação do DNA , Humanos , Íons , Peroxidação de Lipídeos/efeitos dos fármacos , Óxido Nítrico/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Selênio/química , Esteroides Heterocíclicos/química , Células U937RESUMO
The discovery of brassinolide in 1979, a milestone in brassinosteroids research, has sparked great interest of brassinolide analogs (BLs) in agricultural applications. Among these BLs, propionylbrassinolide has captured considerable attention because it shows plant growth regulating activity with an excellent durability. Two impurities of propionylbrassinolide were isolated and purified by semi-preparative high-performance liquid chromatography (HPLC), and the chemical structures were confirmed. For simultaneous separation and determination of propionylbrassinolide and impurities, an efficient analytical method based on HPLC with evaporative light scattering detector (HPLC-ELSD) was developed. The optimized analysis was performed on a C18 reversed phase column (250 mm × 4.60 mm, 5 µm) with isocratic elution of acetonitrile and water (90:10, v/v) as the mobile phase. The drift tube temperature of the ELSD system was set to 50 °C and the auxiliary gas pressure was 150 kPa. The regression equations demonstrated a good linear relationship (R² = 0.9989-0.9999) within the test ranges. The limits of detection (LODs) and quantification (LOQs) for propionylbrassinolide, impurity 1 and 2 were 1.3, 1.2, 1,3 and 4.3, 4.0, 4.2 mg/L, respectively. The fully validated HPLC-ELSD method was readily applied to quantify the active ingredient and impurities in propionylbrassinolide technical concentrate. Moreover, the optimized separation conditions with ELSD have been successfully transferred to mass spectrometry (MS) detector for LC-MS determination.
Assuntos
Brassinosteroides/química , Cromatografia Líquida de Alta Pressão , Espalhamento de Radiação , Esteroides Heterocíclicos/química , Limite de Detecção , Espectrometria de Massas , Estrutura Molecular , Reprodutibilidade dos TestesRESUMO
DWARF1 (DWF1) is a sterol C-24 reductase that catalyses the conversion of 24-methylenecholesterol (24-MCHR) to campesterol (CR) in Arabidopsis. A loss-of-function mutant, dwf1, showed similar phenotypic abnormalities to brassinosteroid (BR)-deficient mutants. These abnormalities were reversed in the wild-type phenotype by exogenous application of castasterone (CS) and brassinolide (BL), but not dolichosterone (DS). Accumulation of DS and decreased CS were found in quantitative analysis of endogenous BRs in dwf1. The enzyme solution prepared from dwf1 was unable to convert 6-deoxoDS to 6-deoxoCS and DS to CS, as seen in either wild-type or 35S:DWF1 transgenic plants. This suggests that DWF1 has enzyme activity not only for a sterol C-24 reductase, but also for a BR C-24 reductase that catalyses C-24 reduction of 6-deoxoDS to 6-deoxoCS and of DS to CS in Arabidopsis. Overexpression of DWF1 in a BR-deficient mutant (det2 35S:DWF1) clearly rescued abnormalities found in det2, indicating that DWF1 functions in biosynthesis of active BRs in Arabidopsis. Expression of DWF1 is down-regulated by application of CS and BL and in a BR-dominant mutant, bes1-D. E-boxes in the putative promoter region of DWF1 directly bind to a BR transcription factor, BES1, implying that DWF1 expression is feedback-regulated by BR signaling via BES1. Overall, biosynthesis of 24-methylene BR is an alternative route for generating CS, which is mediated and regulated by DWF1 in Arabidopsis.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Brassinosteroides/biossíntese , Regulação da Expressão Gênica de Plantas , Oxirredutases/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Vias Biossintéticas , Brassinosteroides/química , Oxirredutases/genética , Esteroides Heterocíclicos/químicaRESUMO
Anthocyanins and proanthocyanidins (PAs) are crucial factors that affect the quality of grapes and the making of wine, which were stimulated by various stimuli and environment factors (sugar, hormones, light, and temperature). The aim of the study was to investigate the influence of exogenous 24-Epibrassinolide (EBR) and light on the mechanism of anthocyanins and PAs accumulation in grape berries. Grape clusters were sprayed with EBR (0.4 mg/L) under light and darkness conditions (EBR + L, EBR + D), or sprayed with deionized water under light and darkness conditions as controls (L, D), at the onset of veraison. A large amount of anthocyanins accumulated in the grape skins and was measured under EBR + L and L treatments, whereas EBR + D and D treatments severely suppressed anthocyanin accumulation. This indicated that EBR treatment could produce overlay effects under light, in comparison to that in dark. Real-time quantitative PCR analysis indicated that EBR application up-regulated the expression of genes (VvCHI1, VvCHS2, VvCHS3, VvDFR, VvLDOX, VvMYBA1) under light conditions. Under darkness conditions, only early biosynthetic genes of anthocyanin biosynthesis responded to EBR. Furthermore, we also analyzed the expression levels of the BR-regulated transcription factor VvBZR1 (Brassinazole-resistant 1) and light-regulated transcription factor VvHY5 (Elongated hypocotyl 5). Our results suggested that EBR and light had synergistic effects on the expression of genes in the anthocyanin biosynthesis pathway.
Assuntos
Antocianinas/biossíntese , Brassinosteroides/metabolismo , Hidroxibenzoatos/química , Proantocianidinas/biossíntese , Esteroides Heterocíclicos/metabolismo , Vitis/metabolismo , Antocianinas/genética , Antocianinas/metabolismo , Brassinosteroides/química , DNA Complementar/química , Expressão Gênica , Regulação da Expressão Gênica de Plantas , Hidroxibenzoatos/metabolismo , Luz , Proteínas de Plantas/metabolismo , Proantocianidinas/genética , Proantocianidinas/metabolismo , RNA/química , Metabolismo Secundário , Esteroides Heterocíclicos/química , Fatores de Transcrição/biossíntese , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Regulação para Cima , VinhoRESUMO
Brassinolide (BL) and castasterone (CS) are the representative members of brassinosteroid class of plant steroid hormone having plant growth promoting activity. In this study, eleven CS analogs bearing a variety of side chains were synthesized to determine the effect of the side chain structures on the BL-like activity. The plant hormonal activity was evaluated in a dwarf rice lamina inclination assay, and the potency was determined as the reciprocal logarithm of the 50% effective dose (ED50) from each dose-response curve. The reciprocal logarithm of ED50 (pED50) was decreased dramatically upon deletion of the C-28 methyl group of CS. The introduction of oxygen-containing groups such as hydroxy, methoxy, and ethoxycarbonyl was also unfavorable to the activity. The pED50 was influenced by the geometry of carbon-carbon double bond between C-24 and C-25 (cis and trans), but the introduction of a fluorine atom at the C-25 position of the double bond did not significantly change the activity. The binding free energy (ΔG) was calculated for all ligand-receptor binding interactions using molecular dynamics, resulting that ΔG is linearly correlated with the pED50.
Assuntos
Colestanóis/química , Reguladores de Crescimento de Plantas/química , Sítios de Ligação , Brassinosteroides/química , Brassinosteroides/metabolismo , Brassinosteroides/farmacologia , Colestanóis/metabolismo , Colestanóis/farmacologia , Ligantes , Simulação de Acoplamento Molecular , Oryza/efeitos dos fármacos , Oryza/crescimento & desenvolvimento , Reguladores de Crescimento de Plantas/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Estrutura Terciária de Proteína , Esteroides Heterocíclicos/química , Esteroides Heterocíclicos/metabolismo , Esteroides Heterocíclicos/farmacologiaRESUMO
Steroids are polycyclic compounds that have a wide range of biological activities. They are bio-synthesized from cholesterol through a series of enzyme-mediated transformations, so they are highly lipophilic and readily enter most cells to interact with intracellular receptors, making them ideal vehicles for targeting a broad array of pathologies. New curative agents for cancers have been developed from several steroidal derivatives. Some biologically important properties of modified steroids are dependent on structural features of the steroid moiety and their side chains. Therefore, chemical derivatization of steroids provides a way to modify their function, and many structure-activity relationships have been confirmed by such synthetic modifications. Several studies demonstrate that steroidal heterocyclic derivatives can be effective in the prevention and treatment of many types of hormone-dependent cancers. The present review is a concise report on steroidal heterocyclic derivatives, with special emphasis on steroid heterocyclic derivatives with 5 membered rings or six-membered rings having interesting therapeutic potential as enzyme inhibitors and cytotoxic drugs to be used as candidates for anti-cancer drug development.
Assuntos
Antineoplásicos/farmacologia , Esteroides Heterocíclicos/farmacologia , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Conformação Molecular , Esteroides Heterocíclicos/síntese química , Esteroides Heterocíclicos/químicaRESUMO
A number of water soluble sulfates of 24-epibrassinolide including the 2α,3α-disulfate and all possible monosulfates were synthesized. The target compounds were isolated in crystalline form as the corresponding sodium salts. Pyridine-sulfur trioxide complex was used as sulfating agent followed by transformation of the resulting pyridinium salts into the sodium sulfates by treatment with NaOH. The control of the regioselectivity was achieved by an appropriate use of acetyl and benzyl protecting groups.
Assuntos
Brassinosteroides/química , Brassinosteroides/síntese química , Acetatos/química , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Estereoisomerismo , Esteroides Heterocíclicos/química , Sulfatos/química , Óxidos de Enxofre/químicaRESUMO
Brassinosteroids (BRs) are steroidal phytohormones that regulate various physiological processes, such as root development and stress tolerance. In the present study, we showed that brassinolide (BL) affects potato root in vitro growth in a dose-dependent manner. Low BL concentrations (0.1 and 0.01 µg/L) promoted root elongation and lateral root development, whereas high BL concentrations (1-100 µg/L) inhibited root elongation. There was a significant (P < 0.05) positive correlation between root activity and BL concentrations within a range from 0.01 to 100 µg/L, with the peak activity of 8.238 mg TTC·g-1 FW·h-1 at a BL concentration of 100 µg/L. Furthermore, plants treated with 50 µg/L BL showed enhanced salt stress tolerance through in vitro growth. Under this scenario, BL treatment enhanced the proline content and antioxidant enzymes' (superoxide dismutase, peroxidase, and catalase) activity and reduced malondialdehyde content in potato shoots. Application of BL maintain K+ and Na+ homeostasis by improving tissue K+/Na+ ratio. Therefore, we suggested that the effects of BL on root development from stem fragments explants as well as on primary root development are dose-dependent and that BL application alleviates salt stress on potato by improving root activity, root/shoot ratio, and antioxidative capacity in shoots and maintaining K+/Na+ homeostasis in potato shoots and roots.
Assuntos
Brassinosteroides/química , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Salinidade , Tolerância ao Sal , Solanum tuberosum/efeitos dos fármacos , Esteroides Heterocíclicos/química , Antioxidantes/química , Biomassa , Catalase/metabolismo , Relação Dose-Resposta a Droga , Peróxido de Hidrogênio/química , Malondialdeído/química , Fenótipo , Caules de Planta/metabolismo , Potássio/química , Prolina , Sódio/química , Cloreto de Sódio/química , Solanum tuberosum/crescimento & desenvolvimento , Superóxido Dismutase/metabolismoRESUMO
Anticancer agents consisting of hybrid molecules are used to improve effectiveness and diminish drug resistance. The current study aimed to introduce newly synthesized hetero-steroids of promising anticancer effects. Besides, the pro-apoptotic effects of new compounds were investigated extensively. Several pyrimidino-, triazolopyrimidino-, pyridazino-, and curcumin-steroid derivatives were synthesized, elucidated and confirmed using the spectral and analytical data. The synthesized hetero-steroids, compounds 9, 10, 11, 12, 13, 14, 15, 18, 20, 21, 22 and 24, were tested for their cytotoxic effects versus human breast cancer cells (MCF-7) using neutral red supravital dye uptake assay. Compound 24 (IC50=18µM) showed more inhibitory influence on MCF-7 growth. Using QRT-PCR (Quantitative real time-polymerase chain reaction), CCND1, Survivin, BCL-2, CDC2, P21 and P53, genes expression levels were investigated. The study results disclose that compounds 4, 7, 18, 24 knocked down the expression levels of CCND1, Survivin, BCL-2 and CDC2. However, P21 and P53 were up-regulated by compounds 21, 22. This study introduced promising pro-apoptotic anticancer agents acting through the modulation of key regulators of apoptosis and cell cycle genes.
Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/metabolismo , Curcumina/farmacologia , Esteroides Heterocíclicos/farmacologia , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Proteína Quinase CDC2 , Curcumina/química , Ciclina D1/metabolismo , Quinases Ciclina-Dependentes/metabolismo , Feminino , Humanos , Células MCF-7 , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Esteroides Heterocíclicos/química , Proteína Supressora de Tumor p53RESUMO
Brassinosteroids (Brs) are a newly recognized group of active steroidal hormones that occur at low concentrations in all plant parts and one of the active and stable forms is 24-epibrassinolide (EBR). We investigated the effect of EBR on tomato (Lycopersicon esculentum Mill.) and its mechanism when seedlings were exposed to low temperature and poor light stress conditions. Leaves of stress-tolerant 'Zhongza9' and stress-sensitive 'Zhongshu4' cultivars were pre-treated with spray solutions containing either 0.1 µM EBR or no EBR (control). The plants were then transferred to chambers where they were exposed to low temperatures of 12 °C/6 °C (day/night) under a low light (LL) level of 80 µmol · m(-2) · s(-1). Exogenous application of EBR significantly increased the antioxidant activity of superoxide dismutase, catalase and peroxidase, and decreased the rate of O2 · (-) formation and H2O2 and malondialdehyde contents. Additionally, the ATP synthase ß subunit content was increased by exogenous hormone application. Based on these results, we conclude that exogenous EBR can elicit synergism between the antioxidant enzyme systems and the ATP synthase ß subunit so that scavenging of reactive oxygen species becomes more efficient. These activities enable plants to cope better under combined low temperature and poor light stresses.