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1.
Molecules ; 26(12)2021 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-34205446

RESUMO

A combination of Fourier transform infrared spectroscopy in attenuated total reflectance geometry (FTIR-ATR) and 2D correlation analysis (2D-COS) was applied here for the first time in order to investigate the temperature-dependent dynamical evolution occurring in a particular type of inclusion complex, based on sulfobutylether-ß-cyclodextrin (SBE-ß-CD) as hosting agent and Coumestrol (7,12-dihydorxcoumestane, Coum), a poorly-soluble active compound known for its anti-viral and anti-oxidant activity. For this purpose, synchronous and asynchronous 2D spectra were calculated in three different wavenumber regions (960-1320 cm-1, 1580-1760 cm-1 and 2780-3750 cm-1) and over a temperature range between 250 K and 340 K. The resolution enhancement provided by the 2D-COS offers the possibility to extract the sequential order of events tracked by specific functional groups of the system, and allows, at the same time, the overcoming of some of the limits associated with conventional 1D FTIR-ATR analysis. Acquired information could be used, in principle, for the definition of an optimized procedure capable to provide high-performance T-sensitive drug carrier systems for different applications.


Assuntos
Cumestrol/química , beta-Ciclodextrinas/química , Antioxidantes/química , Antivirais/química , Portadores de Fármacos/química , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Temperatura
2.
Int J Mol Sci ; 22(9)2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-34062716

RESUMO

Non-alcoholic fatty liver disease (NAFLD) is considered the most common liver disorder, affecting around 25% of the population worldwide. It is a complex disease spectrum, closely linked with other conditions such as obesity, insulin resistance, type 2 diabetes mellitus, and metabolic syndrome, which may increase liver-related mortality. In light of this, numerous efforts have been carried out in recent years in order to clarify its pathogenesis and create new prevention strategies. Currently, the essential role of environmental pollutants in NAFLD development is recognized. Particularly, endocrine-disrupting chemicals (EDCs) have a notable influence. EDCs can be classified as natural (phytoestrogens, genistein, and coumestrol) or synthetic, and the latter ones can be further subdivided into industrial (dioxins, polychlorinated biphenyls, and alkylphenols), agricultural (pesticides, insecticides, herbicides, and fungicides), residential (phthalates, polybrominated biphenyls, and bisphenol A), and pharmaceutical (parabens). Several experimental models have proposed a mechanism involving this group of substances with the disruption of hepatic metabolism, which promotes NAFLD. These include an imbalance between lipid influx/efflux in the liver, mitochondrial dysfunction, liver inflammation, and epigenetic reprogramming. It can be concluded that exposure to EDCs might play a crucial role in NAFLD initiation and evolution. However, further investigations supporting these effects in humans are required.


Assuntos
Disruptores Endócrinos/toxicidade , Poluentes Ambientais/toxicidade , Metabolismo dos Lipídeos/efeitos dos fármacos , Hepatopatia Gordurosa não Alcoólica/metabolismo , Compostos Benzidrílicos/toxicidade , Cumestrol/toxicidade , Dioxinas/toxicidade , Disruptores Endócrinos/classificação , Genisteína/toxicidade , Humanos , Hepatopatia Gordurosa não Alcoólica/induzido quimicamente , Hepatopatia Gordurosa não Alcoólica/patologia , Fenóis/toxicidade , Fitoestrógenos/toxicidade , Bifenilos Policlorados/toxicidade
3.
Biomolecules ; 11(2)2021 02 12.
Artigo em Inglês | MEDLINE | ID: mdl-33673122

RESUMO

Coumestrol is a phytoestrogen widely known for its anti-diabetic, anti-oxidant, and anti-inflammatory properties. Thus, it gets a lot of attention as a potential agent in the nutritional therapy of diseases such as obesity and type 2 diabetes. In our study, we evaluated whether coumestrol affects insulin resistance development via the sphingolipid signaling pathway in primary rat hepatocytes. The cells were isolated from the male Wistar rat's liver with the use of collagenase perfusion. Next, we incubated the cells with the presence or absence of palmitic acid and/or coumestrol. Additionally, some groups were incubated with insulin. The sphingolipid concentrations were assessed by HPLC whereas the expression of all the proteins was evaluated by Western blot. Coumestrol markedly reduced the accumulation of sphingolipids, namely, ceramide and sphinganine through noticeable inhibition of the ceramide de novo synthesis pathway in insulin-resistant hepatocytes. Moreover, coumestrol augmented the expression of fatty acid transport proteins, especially FATP5 and FAT/CD36, which also were responsible for excessive sphingolipid accumulation. Furthermore, coumestrol altered the sphingolipid salvage pathway, which was observed as the excessive deposition of the sphingosine-1-phosphate and sphingosine. Our study clearly showed that coumestrol ameliorated hepatic insulin resistance in primary rat hepatocytes. Thus, we believe that our study may contribute to the discovery of novel preventive and therapeutic methods for metabolic disorders.


Assuntos
Cumestrol/farmacologia , Hepatócitos/efeitos dos fármacos , Resistência à Insulina , Transdução de Sinais/efeitos dos fármacos , Esfingolipídeos/metabolismo , Animais , Ácidos Graxos/metabolismo , Hepatócitos/metabolismo , Masculino , Oxirredução , Ratos , Ratos Wistar
4.
Aging (Albany NY) ; 13(4): 5342-5357, 2021 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-33536350

RESUMO

Diabetes-induced oxidative stress is vital in initiating neuronal damage in the diabetic retina, leading to diabetic retinopathy (DR). This study investigates the possible effects of coumestrol (CMS) on streptozotocin (STZ)-induced DR. First, we established a rat model of DR by STZ injection and a cell model involving high-glucose (HG) exposure of human retinal microvascular endothelial cells (hRMECs). We characterized the expression patterns of oxidative stress indicators, pro-inflammatory cytokines, and pro-apoptotic proteins in hRMECs. Polymerase chain reaction showed sirtuin 1 (SIRT1) to be poorly expressed in the retinal tissues of STZ-treated rats and HG-exposed hRMECs, but its expression was upregulated upon treatment with CMS treatment. Furthermore, CMS treatment attenuated the STZ-induced pathologies such as oxidative stress, inflammation, and cell apoptosis. Consistent with the in vivo results, CMS activated the expression of SIRT1, thereby inhibiting oxidative stress, inflammation, and apoptosis of HG-treated hRMECs. From these findings, we concluded that CMS ameliorated DR by inhibiting inflammation, apoptosis and oxidative stress through activation of SIRT1.


Assuntos
Apoptose/efeitos dos fármacos , Cumestrol/farmacologia , Retinopatia Diabética/metabolismo , Células Endoteliais/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Fitoestrógenos/farmacologia , Retina/efeitos dos fármacos , Sirtuína 1/efeitos dos fármacos , Animais , Diabetes Mellitus Experimental/metabolismo , Retinopatia Diabética/patologia , Modelos Animais de Doenças , Células Endoteliais/metabolismo , Glucose/toxicidade , Humanos , Inflamação/metabolismo , Ratos , Retina/metabolismo , Retina/patologia , Vasos Retinianos/citologia , Sirtuína 1/metabolismo
5.
IET Nanobiotechnol ; 14(7): 574-583, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33010132

RESUMO

The authors tested the efficacy of two salt nanoparticles (NPs), namely, copper dioxide (CuO) and tri-calcium phosphate [Ca3(PO4)2] to induce resistance in green bean pods against grey mould and white rot diseases caused by Botrytis cinerea and Sclerotinia sclerotiorum, respectively. High amounts of phytoalexins, kievitone, coumestrol, phaseollidin, 6-ά-hydroxyphaseollin, and phaseollin, were detected in naturally infected and artificially inoculated green bean pods in response to the tested NPs. Green bean plants treated in the field with CuO and Ca3(PO4)2 NPs had the highest mRNA quantity of all the studied defence genes, receptor-like kinase (PvRK20), pathogenesis-related protein (PR1), 1,3-ß-D-glucanase (pvgluc), polygalacturonase inhibitor protein (PvGIP), and alpha-dioxygenase (a-DOX) than that of the control group. CuO NPs followed by Ca3(PO4)2 NPs at 0.15 mg ml-1 were the most potent in increasing the transcriptomic levels of pk20, DOX, PR1, PvGIP, and pvgluc. Field applications of both chemical elicitor NPs exhibited a non-genotoxic effect on the Paulista green bean DNA using eight ISSR primers. The field application of the studied NPs could effectively extend the shelf life of green bean pods by up to 21 days at 7 ± 1°C during marketing and export due to its potent effect against grey mould and white rot diseases.


Assuntos
Ascomicetos , Botrytis , Fabaceae/metabolismo , Fabaceae/microbiologia , Nanopartículas/química , Transcriptoma , Agricultura , Temperatura Baixa , Cobre/química , Cumestrol/análise , DNA/química , Primers do DNA/química , DNA de Plantas/química , Fungos , Perfilação da Expressão Gênica , Isoflavonas/análise , Microscopia Eletrônica de Transmissão , Mutagênicos , Tamanho da Partícula , Doenças das Plantas , Pterocarpanos/análise , Sesquiterpenos/análise , Software , Temperatura
6.
Free Radic Res ; 54(8-9): 629-639, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32924662

RESUMO

Doxorubicin (DOX) acts as the cornerstone in multiple tumour chemotherapy regimens, however, its clinical application is often impeded due to the induction of a severe cardiotoxicity that eventually provokes left ventricular dysfunction and congestive heart failure. Coumestrol (CMT) is a common dietary phytoestrogen with pleiotropic pharmacological effects. The present study aims to investigate the role and mechanism of CMT on DOX-induced cardiotoxicity. Mice were intragastrically administrated with CMT (5 mg/kg/day) for consecutive 2 weeks and then received a single intraperitoneal injection of DOX (15 mg/kg) to mimic the clinical toxic effects after 8-day additional feeding. To verify the role of 5' AMP-activated protein kinase alpha (AMPKα), AMPKα2 global knockout mice were used. H9C2 cells were cultured to further validate the beneficial role of CMT in vitro. CMT administration notably ameliorated oxidative damage, cell apoptosis and cardiac dysfunction in DOX-treated mice. Besides, we observed that DOX-induced reactive oxygen species overproduction and cardiomyocyte apoptosis were also reduced by CMT incubation in H9C2 cells. Mechanistically, CMT activated AMPKα and Ampkα deficiency abolished the beneficial effects of CMT in vivo and in vitro. Finally, we proved that protein kinase A (PKA) was required for CMT-mediated AMPKα activation and cardioprotective effects. CMT activated PKA/AMPKα pathway to alleviate DOX-induced oxidative damage, cell apoptosis and cardiac dysfunction. Our findings provide a promising therapeutic agent for cancer patients receiving anthracycline chemotherapy.


Assuntos
Proteínas Quinases Ativadas por AMP/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Cardiotoxicidade/tratamento farmacológico , Cardiotoxicidade/etiologia , Cumestrol/uso terapêutico , Doxorrubicina/efeitos adversos , Fitoestrógenos/uso terapêutico , Animais , Cardiotoxicidade/patologia , Cumestrol/farmacologia , Masculino , Camundongos , Fitoestrógenos/farmacologia
7.
Adv Biosyst ; 4(4): e1900187, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32293160

RESUMO

Isoflavones are a class of flavonoids present in legumes and are called phytoestrogens because of their estrogen-like activity. Endogenous estrogen is well known to regulate mammary gland morphogenesis during pregnancy. Each isoflavone also has different physiological activities. However, it is difficult to investigate the direct effect of each isoflavone in mammary morphogenesis in vivo because isoflavones are metabolized into different isoflavones by enteric bacteria. In this study, investigated are the direct influences of coumestrol, daidzein, and genistein on mammary structure development and future milk production ability of mammary epithelial cells (MECs) using in vitro culture models. Mouse MECs are cultured in Matrigel with basic fibroblast growth factor and epidermal growth factor to induce ductal branching and alveolar formation, respectively. Coumestrol and genistein inhibit ductal branching and alveolar formation by affecting the proliferation and migration of MECs with the induction of apoptosis. Daidzein hardly influences mammary structure development. Furthermore, pretreatment with coumestrol adversely affects the induction of milk production ability of MECs. These results suggest that each isoflavone differentially influences mammary morphogenesis and future milk production by affecting MEC behaviors. These results also suggest that the culture models are effective to study mammary epithelial morphogenesis in vitro.


Assuntos
Apoptose/efeitos dos fármacos , Cumestrol/efeitos adversos , Células Epiteliais/metabolismo , Genisteína/efeitos adversos , Lactação/efeitos dos fármacos , Glândulas Mamárias Animais/metabolismo , Animais , Cumestrol/farmacologia , Células Epiteliais/patologia , Feminino , Genisteína/farmacologia , Glândulas Mamárias Animais/patologia , Camundongos , Camundongos Endogâmicos ICR
8.
Daru ; 28(1): 97-108, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31912375

RESUMO

BACKGROUND: Phytoestrogens are plant compounds that are structurally similar to estrogen and that possess anti-cancer properties. Previous studies have reported that coumestrol, daidzein and genistein could induce cell death by reducing Annexin A1 protein in leukemic cell lines. Annexin A1 (ANXA1) is involved in cell progression, metastasis, and apoptosis in several types of cancer cells. The present study sought to investigate if the effects of phytoestrogens on apoptosis, cell cycle arrest and phagocytosis in ANXA1-knockdown leukemic cells are mediated through ANXA1 or occurred independently. METHODS: Transfection of ANXA1 siRNA was conducted to downregulate ANXA1 expression in Jurkat, K562 and U937 cells. Apoptosis and cell cycle assays were conducted using flow cytometry. Western blot was performed to evaluate ANXA1, caspases and Bcl-2 proteins expression. Phagocytosis was determined using hematoxylin and eosin staining. RESULTS: The expression of ANXA1 after the knockdown was significantly downregulated in all cell lines. Genistein significantly induced apoptosis associated with an upregulation of procaspase-3, -9, and - 1 in Jurkat cells. The Bcl-2 expression showed no significant difference in Jurkat, K562 and U937 cells. Treatment with phytoestrogens increased procaspase-1 expression in Jurkat and U937 cells while no changes were detected in K562 cells. Flow cytometry analysis demonstrated that after ANXA1 knockdown, coumestrol and genistein caused cell cycle arrest at G2/M phase in selected type of cells. The percentage of phagocytosis and phagocytosis index increased after the treatment with phytoestrogens in all cell lines. CONCLUSION: Phytoestrogens induced cell death in ANXA1-knockdown leukemia cells, mediated by Annexin A1 proteins. Graphical abstract.


Assuntos
Anexina A1/genética , Cumestrol/farmacologia , Genisteína/farmacologia , Isoflavonas/farmacologia , Fitoestrógenos/farmacologia , Anexina A1/metabolismo , Apoptose/efeitos dos fármacos , Caspases/metabolismo , Ciclo Celular/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Humanos , Células Jurkat , Células K562 , Leucemia/genética , Leucemia/metabolismo , Fagocitose/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Interferente Pequeno/genética , Células THP-1 , Células U937
9.
Toxicol Sci ; 173(1): 19-31, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31626307

RESUMO

The present study assessed the potential of a generic physiologically based kinetic (PBK) model to convert in vitro data for estrogenicity to predict the in vivo uterotrophic response in rats for diethylstibestrol (DES), ethinylestradiol (EE2), genistein (GEN), coumestrol (COU), and methoxychlor (MXC). PBK models were developed using a generic approach and in vitro concentration-response data from the MCF-7 proliferation assay and the yeast estrogen screening assay were translated into in vivo dose-response data. Benchmark dose analysis was performed on the predicted data and available in vivo uterotrophic data to evaluate the model predictions. The results reveal that the developed generic PBK model adequate defines the in vivo kinetics of the estrogens. The predicted dose-response data of DES, EE2, GEN, COU, and MXC matched the reported in vivo uterus weight response in a qualitative way, whereas the quantitative comparison was somewhat hampered by the variability in both in vitro and in vivo data. From a safety perspective, the predictions based on the MCF-7 proliferation assay would best guarantee a safe point of departure for further risk assessment although it may be conservative. The current study indicates the feasibility of using a combination of in vitro toxicity data and a generic PBK model to predict the relative in vivo uterotrophic response for estrogenic chemicals.


Assuntos
Bioensaio/métodos , Estrogênios/toxicidade , Útero/fisiologia , Animais , Cumestrol/toxicidade , Dietilestilbestrol/toxicidade , Relação Dose-Resposta a Droga , Estrona , Etinilestradiol/toxicidade , Feminino , Genisteína/toxicidade , Cinética , Metoxicloro/toxicidade , Modelos Biológicos , Fenóis , Ratos , Útero/efeitos dos fármacos
10.
Cancer Epidemiol Biomarkers Prev ; 29(2): 500-508, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31826911

RESUMO

BACKGROUND: Very few previous studies have examined the relationship between thyroid cancer risk and intake of phytoestrogens (PE); furthermore, these studies have reached inconsistent results. METHODS: We analyzed data from a population-based case-control study in Connecticut from 2010 to 2011, including 387 histologically confirmed thyroid cancer cases and 433 population-based controls, with compound data available concerning specific PEs. Multivariate unconditional logistic regression models were used to estimate the associations between specific PEs and the risk of thyroid cancer, adjusting for potential confounders. RESULTS: An elevated risk of thyroid cancer was associated with moderate to high levels of coumestrol intake [OR = 2.48, 95% confidence interval (CI), 1.39-4.43 for 40-80 µg/day; OR = 2.41, 95% CI, 1.32-4.40 for 80-130 µg/day; and OR = 2.38, 95% CI, 1.26-4.50 for >200 µg/day compared with <40 µg/day], and the main elevation in risk appeared among microcarcinomas (≤1 cm). A decreased risk of papillary macrocarcinomas (>1 cm; OR = 0.26, 95% CI, 0.08-0.85 for 1,860-3,110 µg/day compared with <760 µg/day) was associated with moderate genistein intake among women. CONCLUSIONS: Our study suggests that high coumestrol intake increases the risk of thyroid cancer, especially microcarcinomas, whereas moderate amounts of genistein intake appear to be protective for females with thyroid macrocarcinomas. IMPACT: The study highlights the importance of distinguishing between microcarcinomas and macrocarcinomas in future research on the etiology of thyroid cancer.


Assuntos
Inquéritos sobre Dietas/estatística & dados numéricos , Comportamento Alimentar , Fitoestrógenos/administração & dosagem , Neoplasias da Glândula Tireoide/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Connecticut/epidemiologia , Cumestrol/administração & dosagem , Cumestrol/efeitos adversos , Feminino , Genisteína/administração & dosagem , Genisteína/efeitos adversos , Humanos , Masculino , Pessoa de Meia-Idade , Fitoestrógenos/efeitos adversos , Fatores de Proteção , Fatores de Risco , Neoplasias da Glândula Tireoide/etiologia , Neoplasias da Glândula Tireoide/prevenção & controle , Adulto Jovem
11.
J Nutr Biochem ; 76: 108300, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31812908

RESUMO

Coumestrol is a dietary phytoestrogen with estrogen-mimicking characteristics. This study investigated the molecular mechanisms of antiobesity effects of coumestrol. Two weeks of coumestrol treatment reduced body weight and improved glucose tolerance of high-fat diet (HFD)-fed mice. Notably, coumestrol treatment reduced adiposity but expanded brown adipose tissue mass. In addition, coumestrol treatment induced up-regulation of brown adipocyte markers and lipolytic gene expression in adipose tissue. Mechanistically, coumestrol induced an increase in mitochondrial contents of brown adipose tissue, which was associated with up-regulation of adenosine monophosphate-activated protein kinase and sirtuin 1. In vitro knockdown of estrogen receptor 1 inhibited the effect of coumestrol on brown adipogenic marker expression, increase in mitochondrial contents and oxygen consumption rate in brown adipocytes. Furthermore, lineage tracing of platelet-derived growth factor receptor A-positive (PDGFRA+) adipocyte progenitors confirmed increased levels of de novo brown adipogenesis from PDGFRA+ cells by coumestrol treatment. In conclusion, our results indicate that coumestrol has antiobesity effects through the expansion and activation of brown adipose tissue metabolism.


Assuntos
Tecido Adiposo Marrom/metabolismo , Cumestrol/farmacologia , Obesidade/tratamento farmacológico , Obesidade/metabolismo , Adipócitos Bege/efeitos dos fármacos , Adipogenia , Tecido Adiposo Marrom/efeitos dos fármacos , Tecido Adiposo Branco/metabolismo , Adiposidade , Animais , Peso Corporal , Dieta Hiperlipídica , Teste de Tolerância a Glucose , Lipólise , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fitoestrógenos/farmacologia
12.
Int J Dev Neurosci ; 79: 86-95, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31693927

RESUMO

INTRODUCTION: Neonatal Hypoxia-Ischemia (HI) is a major cause of morbidity and mortality, and is frequently associated with short and long-term neurologic and cognitive impairments. The HI injury causes mitochondrial damage leading to increased production of reactive oxygen species (ROS). Phytoestrogens are non-steroidal plant substances structurally and functionally similar to estrogen. Coumestrol is a potent isoflavonoid with a protective effect against ischemic brain damage in adult rats. Our aim was to determine if coumestrol treatment following neonatal HI attenuates the long-term cognitive deficits induced by neonatal HI, as well as to investigate one possible mechanism underlying its potential effect. METHODS: On the 7th postnatal day, male Wistar rats were submitted to the Levine-Rice HI model. Intraperitoneal injections of 20 mg/kg of coumestrol, or vehicle, were administered immediately pre-hypoxia or 3 h post-hypoxia. At 12 h after HI the mitochondrial status and ROS levels were determined. At 60th postnatal day the cognitive deficits were revealed in the Morris water maze reference and working spatial memories. Following behavioral analysis, histological assessment was performed and reactive astrogliosis was measured by GFAP expression. RESULTS: Results demonstrate that both pre- and post-HI administration of coumestrol were able to counteract the long-term cognitive and morphological impairments caused by HI, as well as to block the late reactive astrogliosis. The pre-HI administration of coumestrol was able to prevent the early mitochondrial dysfunction in the hippocampus of injured rat pups. CONCLUSION: Present data suggest that coumestrol exerts protection against experimental neonatal brain hypoxia-ischemia through, at least in part, early modulation of mitochondrial function.


Assuntos
Encéfalo/efeitos dos fármacos , Cognição/efeitos dos fármacos , Disfunção Cognitiva/tratamento farmacológico , Cumestrol/farmacologia , Hipóxia-Isquemia Encefálica/tratamento farmacológico , Mitocôndrias/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Animais , Encéfalo/metabolismo , Disfunção Cognitiva/metabolismo , Cumestrol/uso terapêutico , Hipóxia-Isquemia Encefálica/metabolismo , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Mitocôndrias/metabolismo , Fármacos Neuroprotetores/uso terapêutico , Fitoestrógenos/farmacologia , Fitoestrógenos/uso terapêutico , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo
13.
J Nat Prod ; 82(4): 1014-1018, 2019 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-30840451

RESUMO

Casein kinase 2 (CK2) is an anti-apoptotic cancer-sustaining protein kinase. Its crystallographic structures with the natural compounds coumestrol, a phytoestrogen, and boldine, an alkaloid, are reported. Coumestrol shows different inhibitory activity against the isolated catalytic α-subunit and the α2ß2 holoenzyme and is able to discriminate between two conformations of the hinge/αD region, whose intrinsic flexibility is a relevant selectivity determinant among kinases. Boldine explores a small cavity at the bottom of the ATP-binding pocket through a local deviation from planarity, a unique case among CK2 inhibitors. The two compounds have different impacts on protein flexibility, which correlate with their different properties.


Assuntos
Trifosfato de Adenosina/metabolismo , Aporfinas/metabolismo , Caseína Quinase II/metabolismo , Cumestrol/metabolismo , Estrutura Molecular
14.
Int J Pharm ; 562: 86-95, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30885651

RESUMO

Coumestrol is a polyphenol with promising therapeutic applications as phytoestrogen, antioxidant and potential cancer chemoprevention agent. The presence of two hydroxyl groups on its chemical structure, with orientation analogous to estradiol, is responsible of both, its antioxidant capacity and its estrogenic activity. However, several studies show that the interaction of polyphenols with food and plasma proteins reduces their antioxidant efficacy. We studied the interaction of coumestrol with bovine serum albumin protein (BSA) by fluorescence spectroscopy and circular dichroism techniques, and the effect of this interaction on its antioxidant activity as a hydroxyl radical scavenger. In addition, coumestrol antioxidant capacity profile using different assays (DPPH, ORAC-FL and ORAC-EPR) was studied. To explain its reactivity we used several methodologies, including DFT calculations, to define its antioxidant mechanism. Coumestrol antioxidant activity unveiled interesting antioxidant properties. BSA interaction with coumestrol reduces significantly photolytic degradation in several media thus preserving its antioxidant properties. Results suggest no significant changes in BSA structure and activity when interacting with coumestrol. Furthermore, this interaction is stronger than for other phytoestrogens such as daidzein and genistein. Considering our promising results, we reported for the first time the fabrication and characterization of coumestrol-loaded albumin nanoparticles. The resulting spherical and homogeneous nanoparticles showed a diameter close to 96 nm. The coumestrol incorporation efficiency in BSA NPs was 22.4%, which is equivalent to 3 molecules of coumestrol for every 10 molecules of BSA.


Assuntos
Antioxidantes/química , Cumestrol/química , Portadores de Fármacos/química , Nanopartículas/química , Fitoestrógenos/química , Soroalbumina Bovina/química , Radical Hidroxila/química
15.
Sci Rep ; 9(1): 1934, 2019 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-30760815

RESUMO

Coumestrol (CMS), a coumestan isoflavone, plays key roles in nodulation through communication with rhizobia, and has been used as phytoestrogens for hormone replacement therapy in humans. Because CMS content is controlled by multiple genetic factors, the genetic basis of CMS biosynthesis has remained unclear. We identified soybean genotypes with consistently high (Daewonkong) or low (SS0903-2B-21-1-2) CMS content over 2 years. We performed RNA sequencing of leaf samples from both genotypes at developmental stage R7, when CMS levels are highest. Within the phenylpropanoid biosynthetic pathway, 41 genes were tightly connected in a functional co-expression gene network; seven of these genes were differentially expressed between two genotypes. We identified 14 candidate genes involved in CMS biosynthesis. Among them, seven were annotated as encoding oxidoreductases that may catalyze the transfer of electrons from daidzein, a precursor of CMS. Two of the other genes, annotated as encoding a MYB domain protein and a MLP-like protein, may increase CMS accumulation in response to stress conditions. Our results will help to complete our understanding of the CMS biosynthetic pathway, and should facilitate development of soybean cultivars with high CMS content that could be used to promote the fitness of plants and human beings.


Assuntos
Vias Biossintéticas/fisiologia , Cumestrol , Regulação da Expressão Gênica de Plantas/fisiologia , RNA-Seq , Soja , Cumestrol/biossíntese , Cumestrol/genética , Perfilação da Expressão Gênica , Soja/genética , Soja/metabolismo
16.
Bioorg Chem ; 85: 140-151, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30612080

RESUMO

There is much interest in the use of phytoestrogens such as coumestrol in breast cancer intervention due to their antiestrogenic activity and multiple modes of tumor cell death. However, the clear beneficial effects of naturally occurring estrogen mimetic coumestrol remain controversial due to experimental evidence that it has been shown to stimulate MCF-7 cell proliferation via agonist effect on estrogen receptor at low concentration. Herein, to disconnect the ER interaction and apoptosis-specific mechanism of coumestrol, various 3, 9-di-O-substituted coumestrols (7a-7e) and their furan ring-opened analogs (5a-5e) were synthesized and assessed for antiproliferative properties. Attachment of a dimethylamine-containing side chain to 3-O of coumestrol led to the most promising compound 7e with improved antiproliferative activity (1.7-fold increase) against MCF-7 cells, decreased estrogen activity (>20 times weaker ERα binder) and a novel action to induce apoptosis. Mechanistic studies revealed that 7e is a tubulin polymerization inhibitor, which could arrest cell cycle at G2/M phase and induce apoptosis along with the decrease of mitochondrial membrane potential. In summary, such subtle modifications to the 3, 9-di-hydroxyl groups of coumestrol allow the generation of a novel apoptosis inducer with distinct pharmacological properties, providing an excellent starting point to future development of novel tumor-vascular disrupting agents targeting tubulin.


Assuntos
Aminas/farmacologia , Inibidores da Angiogênese/farmacologia , Apoptose/efeitos dos fármacos , Cumestrol/análogos & derivados , Cumestrol/farmacologia , Aminas/síntese química , Aminas/metabolismo , Inibidores da Angiogênese/síntese química , Inibidores da Angiogênese/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cumestrol/metabolismo , Receptor alfa de Estrogênio/metabolismo , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Moduladores de Tubulina/síntese química , Moduladores de Tubulina/metabolismo , Moduladores de Tubulina/farmacologia
17.
Environ Sci Pollut Res Int ; 25(32): 32346-32357, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30229492

RESUMO

Advanced oxidation processes have become increasingly important to treat non-biodegradable compounds entering environmental waters. In recent decades, water-soluble metallophthalocyanines have been shown to catalyse H2O2-containing oxidation reactions through the production of unique reactive species, nucleophilic metal-peroxo complexes. Few reports in the literature have examined water insoluble metallophthalocyanines (MPc). The oxidative catalytic activity of water insoluble manganese- and iron-phthalocyanine (MnPc, FePc) at pH 7 has been shown through the decolourisation of methylene blue and removal of bisphenol A. These studies expand on this previous study, exploring the catalytic activity of a range of metallophthalocyanines catalysts under both acidic and neutral conditions. FePc, while only active under neutral conditions, was the best performing catalyst. This activity was significantly improved upon by the addition of acetonitrile as a co-solvent, as well as increasing the ratio of H2O2 to catalyst. MnPc was catalytically active at both pH 3 and 7. FePc and MnPc catalysts showed the ability to remove bisphenol A in the presence of dam water. Reaction rates were reduced for bisphenol A removal with FePc as a catalyst but were unchanged in the presence of MnPc. The removal of 17ß-estradiol, estrone, and coumestrol was successfully demonstrated, with greater than 96% removal of all tested EDC's achieved. This is the first reported study showing the removal of the phytoestrogen, coumestrol. Even though considerably lower concentrations of costly catalysts and oxidation reagents were used in our work, the removal extent of EDC's by the MPc-catalysed oxidation reactions achieved here compares favourably with literature.


Assuntos
Disruptores Endócrinos , Peróxido de Hidrogênio/química , Indóis/química , Ferro/química , Manganês/química , Compostos Organometálicos/química , Purificação da Água/métodos , Compostos Benzidrílicos/análise , Catálise , Cumestrol/análise , Disruptores Endócrinos/análise , Estradiol/análise , Estrona/análise , Compostos Ferrosos/química , Concentração de Íons de Hidrogênio , Oxidantes/química , Oxirredução , Fenóis/análise , Fitoestrógenos/análise , Poluentes Químicos da Água/análise
18.
J Pharm Biomed Anal ; 161: 129-135, 2018 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-30149188

RESUMO

A combination of in vitro and in silico approaches was employed to investigate the estrogenic activities of flavonoid compounds from Psoralea corylifolia. In order to develop fluorescence polarization (FP) assay for flavonoids, a soluble recombinant protein human estrogen receptor α ligand binding domain (hERα-LBD) was produced in Escherichia coli strain. The competition binding experiment was performed by using coumestrol (CS) as a tracer. The result of FP assay suggested that the tested flavonoids can bind to hERα-LBD as affinity ligands, except for corylin. Then, molecular modeling was conducted to explore the binding modes between hERα-LBD and flavonoids. All the tested compounds fit into the hydrophobic binding pocket of hERα-LBD. The hydrophobic and hydrogen-bonding interactions are dominant forces to stabilize the flavonoids-hERα-LBD binding. It can be speculated from molecular docking study that the hydroxyl groups and prenyl group are essential for flavonoid compounds to possess estrogenic activities. Both methylation of hydroxyl group and cyclization of prenyl group significantly diminish the estrogenic potency of flavonoids. Furthermore, quantitative structure-activity relationship (QSAR) analysis was performed by the calculated binding energies of flavonoids coupled with their determined binding affinities. Comparison between the docking scores and the pIC50 values yields an R-squared value of 0.9722, indicating that the estrogenic potency of flavonoids is structure-dependent. In conclusion, molecular docking can potentially be applied for predicting the receptor-binding properties of undescribed compounds based on their molecular structure.


Assuntos
Receptor alfa de Estrogênio/efeitos dos fármacos , Estrogênios/química , Estrogênios/farmacologia , Flavonoides/química , Flavonoides/farmacologia , Polarização de Fluorescência/métodos , Psoralea/química , Ligação Competitiva/efeitos dos fármacos , Cumestrol/farmacologia , Ligação de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Modelos Moleculares , Simulação de Acoplamento Molecular , Estrutura Molecular , Relação Quantitativa Estrutura-Atividade
19.
Anal Chim Acta ; 1032: 107-113, 2018 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-30143207

RESUMO

A fluorescence polarization (FP) assay based on estrogen receptor was developed for the determination of bisphenol compounds (BPs). The human estrogen receptor α ligand binding domain (hERα-LBD) and coumestrol were employed as recognition element and fluorescent probe, respectively. Competitive displacement of tracer from receptor suggested that BPs exhibited dose-dependent binding to hERα-LBD. In order to elucidate the structural basis for the interaction between BPs and hERα-LBD, molecular dynamics simulations were performed to explore their complexation mechanism. The docked bisphenol compounds adopted agonist/antagonist conformations with varying positions and orientations in the hydrophobic binding pocket, depending on their structural characteristics of bridging moieties. Interestingly, the calculated binding energies were generally correlated with the experimentally measured affinities, indicating a potential advantage of the molecular modeling approach in predicting the binding potencies of putative ligands. Considering that the real samples may contain more than one BP, the established FP assay can potentially be used as a pre-screening method to determine the total amounts of bisphenol compounds.


Assuntos
Compostos Benzidrílicos/análise , Cumestrol/química , Polarização de Fluorescência , Corantes Fluorescentes/química , Fenóis/análise , Receptores de Estrogênio/química , Humanos , Modelos Moleculares
20.
Biomed Pharmacother ; 102: 1015-1024, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29710518

RESUMO

Prenylation of bioactive natural compounds has been postulated to be able to enhance the utilization rate and affinity of the compounds with cell membranes, thus promote their bioactivities. Coumestrol, isolated from Medicago sativa, has been known as a phytoestrogen which has bone health benefits. In our previous work, psoralidin, a prenylated coumestrol, was proved to have a higher ability than coumestrol to promote bone formation and to attenuate resorption in vitro. However, it remains to be investigated whether psoralidin will have stronger bone health benefits than coumestrol. In the current study, psoralidin was isolated from Psoralea corylifolia L. and the osteotropic activities of coumestrol and psoralidin were compared in ovariectomized (OVX) rats. Both coumestrol and psoralidin were found to suppress OVX-induced bone loss in vivo, as shown by improved total bone mineral content (t-BMC) or density (t-BMD) and mineral apposition rate, bone biomechanical properties, microstructure and trabecular bone formation, enhanced osteogenic differentiation but suppressed adipogenic differentiation of bone marrow stromal cells (BMSCs), and activation of PI3K/Akt axis and downstream factors such as GSK3ß/ß-catenin and Nrf-2/HO-1. However, psoralidin was shown to have higher activities than coumestrol in the above measurements/indices. Our findings demonstrate that psoralidin, as a novel anti-osteoporosis candidate, could suppress bone loss in OVX rats and have better osteoprotective effects than coumestrol, which may be related to the presence of the isopentenyl group in psoralidin.


Assuntos
Benzofuranos/farmacologia , Cumarínicos/farmacologia , Cumestrol/química , Cumestrol/farmacologia , Osteogênese/efeitos dos fármacos , Pentanos/química , Fosfatidilinositol 3-Quinases/metabolismo , Substâncias Protetoras/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Adipogenia/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Benzofuranos/química , Biomarcadores/sangue , Biomarcadores/urina , Fenômenos Biomecânicos/efeitos dos fármacos , Densidade Óssea/efeitos dos fármacos , Remodelação Óssea/efeitos dos fármacos , Calcificação Fisiológica , Cumarínicos/química , Estradiol/farmacologia , Feminino , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Minerais/metabolismo , Osteoclastos/efeitos dos fármacos , Osteoclastos/enzimologia , Ovariectomia , Oxirredução , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Fosfatase Ácida Resistente a Tartarato/metabolismo , Útero/efeitos dos fármacos , Ganho de Peso/efeitos dos fármacos
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