RESUMO
In Daphnia magna, 20-hydroecdysone (20E) is the main molting hormone and its metabolism is of interest to identify new biomarkers of exposure to contaminants. The present study aimed to (i) assess baseline levels of 20E and transcription levels of four related-genes (shade, neverland, ultraspiracle, and ecdysteroid receptor); and (ii) evaluate effects in D. magna after 21 days of exposure to fenarimol (anti-ecdysteroid) and a mixture of gemfibrozil and clofibric acid (lipid-lowering drugs) at sublethal concentrations. Endpoints included transcription of the target genes and quantification of 20E, mortality, and reproduction of daphnids. Baseline results showed that average responses were relatively similar and did not vary more than 2-fold. However, intra-day variation was generally high and could be explained by sampling individuals with slightly different stages of their development. Exposure tests indicated a significant decrease in daphnid reproduction following chronic exposure to a concentration of 565 µg/L of fenarimol. However, no difference was observed between the control and exposed groups for any of the investigated genes, nor for the levels of 20E after 21 days of exposure. Following exposition to gemfibrozil and clofibric acid at 1 µg/L, no changes were observed for the measured parameters. These results suggest that changes in transcription levels of the target genes and concentrations of 20E may not be sensitive endpoints that can be used as biomarkers of sublethal exposure to the target compounds in D. magna. Measuring multiple time points instead of a single measure as well as additional molecular endpoints obtained from transcriptomic and metabolomic studies could afford more insights on the changes occurring in exposed daphnids to lipid-altering compounds and identify efficient biomarkers of sublethal exposure.
Assuntos
Ecdisterona , Poluentes Químicos da Água , Humanos , Animais , Ecdisterona/metabolismo , Ecdisterona/farmacologia , Muda/genética , Genfibrozila/toxicidade , Reprodução , Biomarcadores/metabolismo , Ácido Clofíbrico/metabolismo , Ácido Clofíbrico/farmacologia , Daphnia , Poluentes Químicos da Água/metabolismoRESUMO
Oxidative stress and inflammation have been considered the main factors in the liver injury of clofibrate (CF). To obtain a novel antihyperlipidemic agent with antioxidant, anti-inflammation and hepatoprotection, the combination of sesamol and clofibric acid moieties was performed and achieved sesamol-clofibrate (CF-Sesamol). CF-Sesamol showed significant hypolipidemia effects in hyperlipidemia mice induced by Triton WR 1339, reducing TG by 38.8% (P < 0.01) and TC by 35.1% (P < 0.01). CF-Sesamol also displayed an alleviating effect on hepatotoxicity. The hepatic weight and hepatic coefficient were decreased. The amelioration of liver function was observed, such as aspartate and lactate transaminases (AST and ALT), alkaline phosphatase (ALP) and total proteins (TP) levels. Liver histopathological examination showed that hepatocyte necrosis, cytoplasmic loosening, nuclear degeneration and inflammatory cell infiltration reduced obviously by treatment with CF-Sesamol. Related molecular mechanisms on hepatoprotection showed that CF-Sesamol up-regulated Nrf2 and HO-1 expression and down-regulated p-NF-κB p65 expression in hepatic tissues. CF-Sesamol has significant antioxidant and anti-inflammatory effects. Plasma antioxidant enzymes such as SOD and CAT increased, anti-lipid peroxidation product MDA decreased. The expression of TNF-α and IL-6 inflammatory cytokines in liver was significantly lower than that in the CF group. The results indicated that CF-Sesamol exerted more potent antihyperlipidemic effects and definite hepatoprotective activity partly through the Nrf2/NF-κB-mediated signaling pathway.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Antioxidantes/farmacologia , Benzodioxóis/farmacologia , Ácido Clofíbrico/farmacologia , Hipolipemiantes/farmacologia , Fenóis/farmacologia , Substâncias Protetoras/farmacologia , Animais , Anti-Inflamatórios não Esteroides/síntese química , Anti-Inflamatórios não Esteroides/química , Antioxidantes/síntese química , Antioxidantes/química , Benzodioxóis/sangue , Benzodioxóis/química , Ácido Clofíbrico/sangue , Ácido Clofíbrico/química , Relação Dose-Resposta a Droga , Hiperlipidemias/induzido quimicamente , Hiperlipidemias/tratamento farmacológico , Hipolipemiantes/síntese química , Hipolipemiantes/química , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Camundongos , Camundongos Endogâmicos , Simulação de Acoplamento Molecular , Estrutura Molecular , Estresse Oxidativo/efeitos dos fármacos , Fenóis/sangue , Fenóis/química , Polietilenoglicóis , Substâncias Protetoras/síntese química , Substâncias Protetoras/química , Relação Estrutura-AtividadeRESUMO
The biogenesis of peroxisomes in relation to the trafficking of proteins to peroxisomes has been extensively examined. However, the supply of phospholipids, which is needed to generate peroxisomal membranes in mammals, remains unclear. Therefore, we herein investigated metabolic alterations induced by clofibric acid, a peroxisome proliferator, in the synthesis of phospholipids, particularly phosphatidylethanolamine (PE) molecular species, and their relationship with the biogenesis of peroxisomal membranes. The subcutaneous administration of clofibric acid to rats at a relatively low dose (130 mg/kg) once a day time-dependently and gradually increased the integrated perimeter of peroxisomes per 100 µm2 hepatocyte cytoplasm (PA). A strong correlation was observed between the content (µmol/mg DNA) of PE containing arachidonic acid (20:4) and PA (r2 = 0.9168). Moreover, the content of PE containing octadecenoic acid (18:1) positively correlated with PA (r2 = 0.8094). The treatment with clofibric acid markedly accelerated the formation of 16:0-20:4 PE by increasing the production of 20:4 and the activity of acyl chain remodeling of pre-existing PE molecular species. Increases in the acyl chain remodeling of PE by clofibric acid were mainly linked to the up-regulated expression of the Lpcat3 gene. On the other hand, clofibric acid markedly increased the formation of palmitic acid (16:0)-18:1 PE through de novo synthesis. These results suggest that the enhanced formation of particular PE molecular species is related to increases in the mass of peroxisomal membranes in peroxisome proliferation in the liver.
Assuntos
Ácido Araquidônico/biossíntese , Ácido Araquidônico/química , Ácido Clofíbrico/farmacologia , Membranas Intracelulares/efeitos dos fármacos , Fígado/citologia , Peroxissomos/efeitos dos fármacos , Fosfatidiletanolaminas/química , Animais , Membranas Intracelulares/metabolismo , Masculino , Peroxissomos/metabolismo , Ratos , Ratos WistarRESUMO
In plants, tolerance to cadmium (Cd) stress is closely related to indole-3-acetic acid (IAA) and hydrogen peroxide (H2O2). However, it is unclear whether Cd-resistant and -sensitive varieties respond differently to Cd stress. In this study, the effects of dimethylthiourea (DMTU, a H2O2 scavenger) and p-chlorophenoxy isobutyric acid (PCIB, an IAA signaling inhibitor) on root growth, endogenous hormones and antioxidant system were investigated to decipher how DMTU and PCIB treatments alleviate the inhibition of root elongation in Cd-resistant (Commander) and -sensitive (Crossfire III) tall fescue varieties under Cd stress. Both varieties subjected to 10 µM Cd treatments for 12 h presented a substantial decrease in root elongation coupled with a reduction in brassinosteroid (BR) and zeatin riboside (ZR) contents, but the changes in IAA and abscisic acid (ABA) contents under Cd stress were opposite in the two varieties. In addition, the H2O2 content and antioxidant enzyme activities significantly increased in both varieties. However, pretreatment with PCIB or DMTU mitigated the inhibition of root elongation caused by Cd, accompanied by the significant changes of aforementioned physiological parameters. PCIB significantly reduced the IAA content in 'Commander', while DMTU significantly increased the IAA content in 'Crossfire III' and effectively relieved the inhibition of root elongation. But both treatments decreased the Cd-induced H2O2 accumulation. These results indicated that DMTU or PCIB can alleviate the Cd-inhibited root elongation in two varieties whose resistance differed under Cd stress, but they presented differences in the response of hormones, especially IAA, which may be due to the different adaptation mechanisms of two varieties in response to Cd stress.
Assuntos
Cádmio/toxicidade , Ácido Clofíbrico/farmacologia , Festuca/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Poluentes do Solo/toxicidade , Tioureia/análogos & derivados , Ácido Abscísico/metabolismo , Adaptação Fisiológica/efeitos dos fármacos , Festuca/crescimento & desenvolvimento , Festuca/metabolismo , Peróxido de Hidrogênio/metabolismo , Ácidos Indolacéticos/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Tioureia/farmacologiaRESUMO
The presence of pharmaceuticals in water sources, including in drinking water (DW), is increasingly being recognized as an emerging and global concern for the environment and public health. Based on the principles of the "One Health" initiative, the present work aims to understand the effects of clofibric acid (CA), a lipid regulator, on the behavior of a selected bacterium isolated from drinking water (DW). Biofilms of the opportunistic pathogen Stenotrophomonas maltophilia were exposed to CA for 12â¯weeksâ¯at 170 and 17000â¯ng/L. The effects of CA were evaluated on planktonic S. maltophilia susceptibility to chlorine and antibiotics (amoxicillin, ciprofloxacin, erythromycin, kanamycin, levofloxacin, oxacillin, spectinomycin, tetracycline and trimethoprim-sulfamethoxazole), biofilm formation, motility, siderophores production and on the adhesion and internalization of the human colon adenocarcinoma cell line (HT-29). It was found that CA did not affect planktonic S. maltophilia tolerance to chlorine exposure. Additionally, no effects were observed on biofilm formation, motility and siderophores production. However, biofilms formed after CA exposure were more tolerant to chlorine disinfection and lower CFU reductions were obtained. Of additional concern was the effect of CA exposure on S. maltophilia increased tolerance to erythromycin. CA exposure also slightly reduced S. maltophilia ability to invade HT-29â¯cells. In conclusion, this work reinforces the importance of studying the effects of non-antibiotic contaminants on the behavior of environmental microorganisms, particularly their role as drivers affecting resistance evolution and selection.
Assuntos
Biofilmes/efeitos dos fármacos , Ciprofloxacina/farmacologia , Ácido Clofíbrico/farmacologia , Stenotrophomonas maltophilia/efeitos dos fármacos , Antibacterianos/farmacologia , Anti-Infecciosos/farmacologia , Água Potável/microbiologia , Tolerância a Medicamentos , Células HT29 , Humanos , Stenotrophomonas maltophilia/patogenicidade , Stenotrophomonas maltophilia/ultraestrutura , VirulênciaRESUMO
There has been a developing technology in algae with pharmaceuticals wastewater. However, the effect and the underlying mechanism of pharmaceuticals on algae are not well understood. To investigate the effect and mechanism of pharmaceuticalson microalgae, four pharmaceuticals of clofibric acid (CLF), ciprofloxacin (CIP), diclofenac (DCF) and carbamazepine (CBZ) on C. pyrenoidosa culture were analyzed. At low concentrations (<10 mg/L), the pharmaceuticals, especially the DCF, exhibited positive effects on both the structure and function of algal cultures; algal growth (i.e., chlorophyll a accumulation, lipid accumulation) and activities of antioxidant enzymes were stimulated. The algal metabolite differences of various DCF concentrations were investigated and a total of 91 substances were identified, whose samples were clustered and clearly separated. The key metabolomics pathway analysis found that the DCF promoted the carbohydrate and fatty acid metabolic pathway in C. pyrenoidosa under relatively low concentrations (<10 mg/L). However, the algae metabolomics pathway was disturbed significantly under the action of a high concentration of DCF (>100 mg/L). The study detected the effects of four pharmaceuticals on C. pyrenoidosa and demonstrated that the usage of metabolomics analysis complemented with DCF could be an effective approach to understand the mechanism of molecular evolution in C. pyrenoidosa for microalgal biomass and bioenergy from wastewater in researches of biological resources.
Assuntos
Chlorella/efeitos dos fármacos , Chlorella/crescimento & desenvolvimento , Chlorella/metabolismo , Preparações Farmacêuticas , Carbamazepina/farmacologia , Catalase/metabolismo , Técnicas de Cultura de Células , Chlorella/citologia , Clorofila A/metabolismo , Ciprofloxacina/farmacologia , Ácido Clofíbrico/farmacologia , Diclofenaco/farmacologia , Relação Dose-Resposta a Droga , Metabolismo dos Lipídeos/efeitos dos fármacos , Metabolômica , Microalgas/efeitos dos fármacos , Tamanho da Partícula , Superóxido Dismutase/metabolismoRESUMO
Apical dominance is the process whereby the shoot tip inhibits the growth of axillary buds along the stem. It has been proposed that the shoot tip, which is the predominant source of the plant hormone auxin, prevents bud outgrowth by suppressing auxin canalization and export from axillary buds into the main stem. In this theory, auxin flow out of axillary buds is a prerequisite for bud outgrowth, and buds are triggered to grow by an enhanced proportional flow of auxin from the buds. A major challenge of directly testing this model is in being able to create a bud- or stem-specific change in auxin transport. Here we evaluate the relationship between specific changes in auxin efflux from axillary buds and bud outgrowth after shoot tip removal (decapitation) in the pea (Pisum sativum). The auxin transport inhibitor 1-N-naphthylphthalamic acid (NPA) and to a lesser extent, the auxin perception inhibitor p-chlorophenoxyisobutyric acid (PCIB), effectively blocked auxin efflux from axillary buds of intact and decapitated plants without affecting auxin flow in the main stem. Gene expression analyses indicate that NPA and PCIB regulate auxin-inducible, and biosynthesis and transport genes, in axillary buds within 3 h after application. These inhibitors had no effect on initial bud outgrowth after decapitation or cytokinin (benzyladenine; BA) treatment. Inhibitory effects of PCIB and NPA on axillary bud outgrowth only became apparent from 48 h after treatment. These findings demonstrate that the initiation of decapitation- and cytokinin-induced axillary bud outgrowth is independent of auxin canalization and export from the bud.
Assuntos
Ácidos Indolacéticos/metabolismo , Pisum sativum/crescimento & desenvolvimento , Reguladores de Crescimento de Plantas/fisiologia , Transporte Biológico/efeitos dos fármacos , Ácido Clofíbrico/farmacologia , Perfilação da Expressão Gênica , Genes de Plantas , Modelos Biológicos , Pisum sativum/efeitos dos fármacos , Pisum sativum/metabolismo , Ftalimidas/farmacologia , Desenvolvimento Vegetal/efeitos dos fármacos , Reguladores de Crescimento de Plantas/metabolismo , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismoRESUMO
Hemoglobin (Hb) modifiers that stereospecifically inhibit sickle hemoglobin polymer formation and/or allosterically increase Hb affinity for oxygen have been shown to prevent the primary pathophysiology of sickle cell disease (SCD), specifically, Hb polymerization and red blood cell sickling. Several such compounds are currently being clinically studied for the treatment of SCD. Based on the previously reported non-covalent Hb binding characteristics of substituted aryloxyalkanoic acids that exhibited antisickling properties, we designed, synthesized and evaluated 18 new compounds (KAUS II series) for enhanced antisickling activities. Surprisingly, select test compounds showed no antisickling effects or promoted erythrocyte sickling. Additionally, the compounds showed no significant effect on Hb oxygen affinity (or in some cases, even decreased the affinity for oxygen). The X-ray structure of deoxygenated Hb in complex with a prototype compound, KAUS-23, revealed that the effector bound in the central water cavity of the protein, providing atomic level explanations for the observed functional and biological activities. Although the structural modification did not lead to the anticipated biological effects, the findings provide important direction for designing candidate antisickling agents, as well as a framework for novel Hb allosteric effectors that conversely, decrease the protein affinity for oxygen for potential therapeutic use for hypoxic- and/or ischemic-related diseases.
Assuntos
Antidrepanocíticos/química , Hemoglobinas/química , Regulação Alostérica/efeitos dos fármacos , Antidrepanocíticos/síntese química , Antidrepanocíticos/farmacologia , Sítios de Ligação , Ácido Clofíbrico/química , Ácido Clofíbrico/farmacologia , Hemoglobinas/metabolismo , Modelos Moleculares , Conformação Molecular , Ligação Proteica , Relação Estrutura-AtividadeRESUMO
PPARα is a ligand activated transcription factor belonging to the nuclear receptor subfamily, involved in fatty acid metabolism in tissues with high oxidative rates such as muscle, heart and liver. PPARα activation is important in steatosis, inflammation and fibrosis in preclinical models of non-alcoholic fatty liver disease identifying a new potential therapeutic area. In this work, three series of clofibric acid analogues conjugated with naphthyl, quinolin, chloroquinolin and anthracenyl scaffolds were synthesized. In an effort to obtain new compounds active as PPARα agonists, these molecules were evaluated for PPARα transactivation activity. Naphthyl and quinolin derivatives showed a good activation of PPARα; noteworthy, optically active naphthyl derivatives activated PPARα better than corresponding parent compound.
Assuntos
Ácido Clofíbrico/análogos & derivados , PPAR alfa/agonistas , Ácido Clofíbrico/farmacologia , HumanosRESUMO
To improve embryogenesis in microspore cultures of kale (Brassica oleracea L. var. acephala DC.), 6-benzylaminopurine (6-BA), naphthaleneacetic acid (NAA), arabinogalactan (AG), p-chlorophenoxyisobutyric acid (PCIB), and activated charcoal (AC) were added to the medium using four varieties of kale. The results showed that the addition of AG (0.1-0.2 g/L), AC (0.1-0.2 g/L) or a combination of 6-BA (0.1-0.2 mg/L) and NAA (0.1-0.2 mg/L) promoted embryo-genesis. Adding 40 µM PCIB or a combination of 40 µM PCIB and 0.2 g/L AC to NLN-13 medium at pH 5.8 effectively enhanced embryogenesis. Treatment with a combination of 40 µM PCIB and 10 mg/L AG gave the highest rate of embryonic induction, especially in genotype "Y007," which showed a twelve-fold increase in yield.
Assuntos
Brassica/embriologia , Carvão Vegetal/farmacologia , Ácido Clofíbrico/farmacologia , Galactanos/farmacologia , Sementes/crescimento & desenvolvimento , Brassica/genética , PloidiasRESUMO
Isolated microspores are reprogrammed in vitro by stress, becoming totipotent cells and producing embryos and plants via a process known as microspore embryogenesis. Despite the abundance of data on auxin involvement in plant development and embryogenesis, no data are available regarding the dynamics of auxin concentration, cellular localization and the expression of biosynthesis genes during microspore embryogenesis. This work involved the analysis of auxin concentration and cellular accumulation; expression of TAA1 and NIT2 encoding enzymes of two auxin biosynthetic pathways; expression of the PIN1-like efflux carrier; and the effects of inhibition of auxin transport and action by N-1-naphthylphthalamic acid (NPA) and α-(p-chlorophenoxy) isobutyric acid (PCIB) during Brassica napus microspore embryogenesis. The results indicated de novo auxin synthesis after stress-induced microspore reprogramming and embryogenesis initiation, accompanying the first cell divisions. The progressive increase of auxin concentration during progression of embryogenesis correlated with the expression patterns of TAA1 and NIT2 genes of auxin biosynthetic pathways. Auxin was evenly distributed in early embryos, whereas in heart/torpedo embryos auxin was accumulated in apical and basal embryo regions. Auxin efflux carrier PIN1-like gene expression was induced in early multicellular embryos and increased at the globular/torpedo embryo stages. Inhibition of polar auxin transport (PAT) and action, by NPA and PCIB, impaired embryo development, indicating that PAT and auxin action are required for microspore embryo progression. NPA also modified auxin embryo accumulation patterns. These findings indicate that endogenous auxin biosynthesis, action and polar transport are required in stress-induced microspore reprogramming, embryogenesis initiation and progression.
Assuntos
Brassica napus/metabolismo , Ácidos Indolacéticos/metabolismo , Proteínas de Plantas/metabolismo , Pólen/embriologia , Transporte Biológico , Vias Biossintéticas/genética , Brassica napus/citologia , Brassica napus/genética , Células Cultivadas , Cromatografia Líquida , Ácido Clofíbrico/farmacologia , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Temperatura Alta , Espectrometria de Massas/métodos , Microscopia Confocal , Microscopia de Interferência , Ftalimidas/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/genética , Pólen/efeitos dos fármacos , Pólen/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sementes/citologia , Sementes/genética , Sementes/metabolismo , Estresse FisiológicoRESUMO
BACKGROUND AND AIMS: Barley (Hordeum vulgare) double mutants Hv-Hd/tw2, formed by hybridization, are characterized by inherited phenotypic instability and by several new features, such as bract/leaf-like structures, long naked gaps in the spike, and a wide spectrum of variations in the basic and ectopic flowers, which are absent in single mutants. Several of these features resemble those of mutations in auxin distribution, and thus the aim of this study was to determine whether auxin imbalances are related to phenotypic variations and instability. The effects of auxin inhibitors and 2,4-D (2,4-dichlorophenoxyacetic acid) on variation in basic and ectopic flowers were therefore examined, together with the effects of 2,4-D on spike structure. METHODS: The character of phenotypic instability and the effects of auxin inhibitors and 2,4-D were compared in callus cultures and intact plants of single homeotic Hv-tw2 and Hv-Hooded/Kap (in the BKn3 gene) mutants and alternative double mutant lines: offspring from individual plants in distal hybrid generations (F9-F10) that all had the same BKn3 allele as determined by DNA sequencing. For intact plants, two auxin inhibitors, 9-hydroxyfluorene-9-carboxylic acid (HFCA) and p-chlorophenoxyisobutyric acid (PCIB), were used. KEY RESULTS: Callus growth and flower/spike structures of the Hv-tw2 mutant differed in their responses to HFCA and PCIB. An increase in normal basic flowers after exposure to auxin inhibitors and a decrease in their frequencies caused by 2,4-D were observed, and there were also modifications in the spectra of ectopic flowers, especially those with sexual organs, but the effects depended on the genotype. Exposure to 2,4-D decreased the frequency of short gaps and lodicule transformations in Hv-tw2 and of long naked gaps in double mutants. CONCLUSIONS: The effects of auxin inhibitors and 2,4-D suggest that ectopic auxin maxima or deficiencies arise in various regions of the inflorescence/flower primordia. Based on the phenotypic instability observed, definite trends in the development of ectopic flower structures may be detected, from insignificant outgrowths on awns to flowers with sterile organs. Phenotypically unstable barley double mutants provide a highly promising genetic system for the investigation of gene expression modules and trend orders.
Assuntos
Ácido 2,4-Diclorofenoxiacético/farmacologia , Hordeum , Ácidos Indolacéticos/antagonistas & inibidores , Mutação , Reguladores de Crescimento de Plantas/farmacologia , Ácidos Carboxílicos/farmacologia , Ácido Clofíbrico/farmacologia , Flores/efeitos dos fármacos , Flores/genética , Flores/crescimento & desenvolvimento , Flores/metabolismo , Hordeum/efeitos dos fármacos , Hordeum/genética , Hordeum/crescimento & desenvolvimento , Fenótipo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Auxin is a master regulator of root growth by modulating its development under the constantly changing environment. Recently, an antagonistic interaction was suggested between SA and IAA signaling. Therefore, the purpose of this work was to analyze and compare the effect of the indole-3-acetic acid (IAA) signaling inhibitor p-chlorophenoxyisobutyric acid (PCIB) and salicylic acid (SA) as a potential IAA signaling inhibitor on the root growth, enzyme activity and reactive oxygen species (ROS) production in Cd- and IAA-treated barley root tips. Exposure of plants to Cd resulted in a more than threefold increase of IAA content in the root apex even 3h after the treatment. In addition, exogenously applied IAA evoked root responses such as root growth inhibition and swelling, ROS generation and activation of lipoxygenase or glutathione peroxidase identical to those induced by Cd. Furthermore, both Cd- and IAA-induced stress responses were markedly reversed by PCIB or SA post-treatment. Similarly to PCIB, SA did not affect the IAA content of root tips, suggesting the action of SA on the IAA signaling pathway in barley roots. SA probably does not alleviate the Cd toxicity in roots, but rather prevents or partially inhibits the root defense response to the presence of Cd through the inhibition of Cd-induced IAA-mediated ROS generation in roots.
Assuntos
Cádmio/toxicidade , Hordeum/efeitos dos fármacos , Ácidos Indolacéticos/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Ácido Salicílico/farmacologia , Ácido Clofíbrico/farmacologia , Hordeum/fisiologia , Lipoxigenase/metabolismo , Meristema/efeitos dos fármacos , Meristema/fisiologia , Proteínas de Plantas/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/fisiologia , Transdução de Sinais/efeitos dos fármacosRESUMO
KEY MESSAGE: Effective microspore embryogenesis in triticale is determined by a specific hormonal homeostasis: low value of IAA/cytokinins, IAA/ABA and cytokinins/ABA ratios as well as proper endogenous/exogenous auxin balance, which favours androgenic structure formation and green plant regeneration ability. The concentration of plant growth regulators (PGRs): auxins (Auxs), cytokinins (CKs) and abscisic acid (ABA) was measured in anthers of eight DH lines of triticale (× Triticosecale Wittm.), and associated with microspore embryogenesis (ME) responsiveness. The analysis was conducted on anthers excised from control tillers at the phase optimal for ME induction and then after ME-initiating tillers treatment (21 days at 4 °C). In control, IAA predominated among Auxs (11-39 nmol g(-1) DW), with IBA constituting only 1 % of total Auxs content. The prevailing isoforms of CKs were cis isomers of zeatin (121-424 pmol g(-1) DW) and zeatin ryboside (cZR, 146-432 pmol g(-1) DW). Surprisingly, a relatively high level (10-64 pmol g(-1) DW) of kinetin (KIN) was detected. Cold treatment significantly changed the levels of all analysed PGRs. The anthers of 'responsive' DH lines contained higher concentrations of IBA, cis and trans zeatin, cZR and ABA, and lower amount of IAA and KIN in comparison with 'recalcitrant' genotypes. However, the effects of exogenous ABA, p-chlorophenoxyisobutyric acid (PCIB) and 2,3,5-triiodobenzoic acid treatments suggest that none of the studied PGRs acts alone in the acquisition of embryogenic competency, which seems to be an effect of concerted PGRs crosstalk. The initiation of ME required a certain threshold level of ABA. A crucial prerequisite for high ME effectiveness was a specific PGRs homeostasis: lower Auxs level in comparison with CKs and ABA, and lower CKs/ABA ratio. A proper balance between endogenous Auxs in anthers and exogenous Auxs supplied by culture media was also essential.
Assuntos
Grão Comestível/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Poaceae/metabolismo , Pólen/metabolismo , Ácido Abscísico/metabolismo , Ácido Abscísico/farmacologia , Transporte Biológico/efeitos dos fármacos , Células Cultivadas , Ácido Clofíbrico/farmacologia , Temperatura Baixa , Citocininas/metabolismo , Citocininas/farmacologia , Grão Comestível/citologia , Grão Comestível/genética , Genótipo , Imuno-Histoquímica , Ácidos Indolacéticos/metabolismo , Ácidos Indolacéticos/farmacologia , Microscopia de Fluorescência , Reguladores de Crescimento de Plantas/farmacologia , Técnicas de Embriogênese Somática de Plantas/métodos , Poaceae/citologia , Poaceae/genética , Pólen/embriologia , Pólen/genética , Análise de Componente Principal , Regeneração/efeitos dos fármacos , Regeneração/genética , Ácidos Tri-Iodobenzoicos/farmacologiaRESUMO
Fibrates have been reported to elevate the hepatic proportion of oleic acid (18:1n-9) through inducing stearoyl-CoA desaturase (SCD). Despite abundant studies on the regulation of SCD in the liver, little is known about this issue in the small intestine. The present study aimed to investigate the effect of clofibric acid on the fatty acid profile, particularly monounsaturated fatty acids (MUFA), and the SCD expression in intestinal mucosa. Treatment of rats with a diet containing 0.5% (w/w) clofibric acid for 7 days changed the MUFA profile of total lipids in intestinal mucosa; the proportion of 18:1n-9 was significantly increased, whereas those of palmitoleic (16:1n-7) and cis-vaccenic (18:1n-7) acids were not changed. Upon the treatment with clofibric acid, SCD was induced and the gene expression of SCD1, SCD2, and fatty acid elongase (Elovl) 6 was up-regulated, but that of Elovl5 was unaffected. Fat-free diet feeding for 28 days increased the proportions of 16:1n-7 and 18:1n-7, but did not effectively change that of 18:1n-9, in intestinal mucosa. Fat-free diet feeding up-regulated the gene expression of SCD1, but not that of SCD2, Elovl6, or Elovl5. These results indicate that intestinal mucosa significantly changes its MUFA profile in response to challenges by clofibric acid and a fat-free diet and suggest that up-regulation of the gene expression of SCD along with Elovl6 is indispensable to elevate the proportion of 18:1n-9 in intestinal mucosa.
Assuntos
Ácido Clofíbrico/farmacologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Estearoil-CoA Dessaturase/metabolismo , Acil-CoA Oxidase , Animais , Dieta com Restrição de Gorduras , Ácidos Graxos/metabolismo , Ácidos Graxos Monoinsaturados/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Mucosa Intestinal/enzimologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Lipídeos/química , Masculino , Oxirredutases/genética , PPAR alfa/genética , Ratos Wistar , Estearoil-CoA Dessaturase/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/genéticaRESUMO
Fibrates are used in biochemical and pharmacological studies as bioactive tools. Nevertheless, most studies have lacked information concerning the concentrations of fibric acids working inside tissues because a simple and sensitive method is not available for their quantitation. This study aimed to develop a simple and sensitive bioanalytical method for the quantitation of clofibric, bezafibric and fenofibric acids in samples of very small portions of tissues. Fibric acids were extracted into n-hexane-ethyl acetate from tissue homogenates (10 mg of liver, kidney or muscle) or serum (100 µL) and were derivatized with 4-bromomethyl-6,7-dimethoxycoumarin, followed by HPLC with fluorescence detection. These compounds were separated isocratically on a reversed phase with acetonitrile-water. Standard analytical curves were linear over the concentration range of 0.2-20 nmol/10 mg of liver. Precision and accuracy were within acceptable limits. Recovery from liver homogenates ranged from 93.03 to 112.29%. This method enabled the quantitation of fibric acids in 10 mg of liver from rats treated with clofibric acid, bezafibric acid or fenofibrate. From these analytical data, it became clear that there was no large difference in ratio of acyl-CoA oxidase 1 (Acox1) mRNA level to fibric acid content in the liver among the three fibric acids, suggesting that these three fibric acids have similar potency to increase expression of the Acox1 gene, which is a target of peroxisome proliferator-activated receptor α. Thus, the proposed method is a simple, sensitive and reliable tool for the quantitation of fibric acids working in vivo inside livers.
Assuntos
Acil-CoA Oxidase/metabolismo , Bezafibrato/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Ácido Clofíbrico/metabolismo , Fenofibrato/análogos & derivados , Ácidos Fíbricos/metabolismo , Fígado/metabolismo , Acil-CoA Oxidase/genética , Animais , Bezafibrato/farmacologia , Ácido Clofíbrico/farmacocinética , Ácido Clofíbrico/farmacologia , Fenofibrato/metabolismo , Fenofibrato/farmacocinética , Fenofibrato/farmacologia , Ácidos Fíbricos/farmacocinética , Ácidos Fíbricos/farmacologia , Masculino , PPAR alfa/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Reprodutibilidade dos TestesRESUMO
In an effort to develop safe and efficacious compounds for the treatment of metabolic disorders, new compounds based on a combination of clofibric acid, the active metabolite of clofibrate, and trans-stilbene, chalcone, and other lipophilic groups were synthesized. They were evaluated for PPARα transactivation activity; all branched derivatives showed an increase of the transcriptional activity of receptor compared to the linear ones. Noteworthy, stilbene and benzophenone branched derivatives activated the PPARα better than clofibric acid.
Assuntos
Chalcona/química , Ácido Clofíbrico/farmacologia , PPAR alfa/agonistas , Estilbenos/química , Linhagem Celular , Chalcona/farmacologia , Ácido Clofíbrico/química , Humanos , Estrutura Molecular , Estilbenos/farmacologia , Relação Estrutura-Atividade , Ativação Transcricional/efeitos dos fármacosRESUMO
Hepatic triacylglycerol (TAG) homeostasis is maintained by carefully regulated balance between its synthesis and disposal. Impairment in this balance causes steatosis. The aims of this study were i) to uncover whether fibrates control TAG concentration through the action of adipose triglyceride lipase (ATGL) and ii) to compare the potency of the effects on ATGL expression and TAG concentration among fenofibrate, bezafibrate, and clofibric acid in the liver of rats. Treatments of rats with the three fibrates induced ATGL and concomitantly decreased hepatic TAG concentration. The upregulation of ATGL was likely mediated through the activation of peroxisome proliferator-activated receptor α. Fibrates also expanded capacity of fatty acid ß-oxidation. Importantly, three fibric acids (fenofibric, bezafibric, and clofibric acids) that are active metabolites formed in the liver exhibited almost the same potency to elevate ATGL expression in vivo, despite the fact that there were considerable differences in this regard among fenofibrate, bezafibrate, and clofibric acid when compared on the basis of their dosage. These results suggest that ATGL represents a potential therapeutic target for ameliorating hepatic steatosis and that fibric acids are promising agents to ameliorate and/or protect against hepatic steatosis.
Assuntos
Bezafibrato/farmacologia , Ácido Clofíbrico/farmacologia , Fenofibrato/farmacologia , Lipase/metabolismo , Fígado/enzimologia , Fígado/metabolismo , Triglicerídeos/metabolismo , Regulação para Cima/efeitos dos fármacos , Animais , Bezafibrato/uso terapêutico , Ácido Clofíbrico/uso terapêutico , Fígado Gorduroso/tratamento farmacológico , Fígado Gorduroso/etiologia , Fígado Gorduroso/prevenção & controle , Fenofibrato/uso terapêutico , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Terapia de Alvo Molecular , PPAR alfa/metabolismo , Ratos , Ratos WistarRESUMO
The genes encoding nuclear receptors comprise multiple 5'untranslated exons, which give rise to several transcripts encoding the same protein, allowing tissue-specific regulation of expression. Both human and mouse peroxisome proliferator activated receptor (PPAR) α genes have multiple promoters, although their function is unknown. Here we have characterised the rat PPARα promoter region and have identified three alternative PPARα transcripts, which have different transcription start sites owing to the utilisation of distinct first exons. Moreover these alternative PPARα transcripts were differentially expressed between adipose tissue and liver. We show that while the major adipose (P1) and liver (P2) transcripts were both induced by dexamethasone, they were differentially regulated by the PPARα agonist, clofibric acid, and leptin. Leptin had no effect on the adipose-specific P1 transcript, but induced liver-specific P2 promoter activity via a STAT3/Sp1 mechanism. Moreover in Wistar rats, leptin treatment between postnatal day 3-13 led to an increase in P2 but not P1 transcription in adipose tissue which was sustained into adulthood. This suggests that the expression of the alternative PPARα transcripts are in part programmed by early life exposure to leptin leading to persistent change in adipose tissue fatty acid metabolism through specific activation of a quiescent PPARα promoter. Such complexity in the regulation of PPARα may allow the expression of PPARα to be finely regulated in response to environmental factors.
Assuntos
Regiões 5' não Traduzidas/genética , Éxons/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Leptina/farmacologia , PPAR alfa/genética , Tecido Adiposo/metabolismo , Animais , Animais Recém-Nascidos , Ácido Clofíbrico/farmacologia , Feminino , Células Hep G2 , Humanos , Fígado/metabolismo , Camundongos , Especificidade de Órgãos , Regiões Promotoras Genéticas/genética , Ratos , Ratos Wistar , Transcrição Gênica/efeitos dos fármacosRESUMO
A series of previously synthesized chiral derivatives of clofibric and phenylacetic acids, acting as dual agonists towards the peroxisome proliferator-activated receptors (PPARs) α and γ, was taken into account, and the efficacy of these compounds was analyzed by means of 2D-, 3D-QSAR and docking studies with the goal to gain deeper insights into the three-dimensional determinants governing ligands selectivity for PPARs. By multiregressional analysis a correlation between the lipophilicity and PPARα activity was found, whereas for PPARγ the correlation was achieved once efficacy was related to the presence of polar groups on agonists scaffold. Docking of these compounds further corroborated this hypothesis, and then provided a valid support for subsequent chemometric analysis and pharmacophore models development for both receptors subtypes. Computational results suggested site directed mutagenesis experiments which confirmed the importance of amino acid residues in PPAR activity, allowing the identification of critical hotspots most likely taking over PPARs selectivity.