RESUMEN

During central nervous system (CNS) development, proper and timely induction of neurite elongation is critical for generating functional, mature neurons, and neuronal networks. Despite the wealth of information on the action of extracellular cues, little is known about the intrinsic gene regulatory factors that control this developmental decision. Here, we report the identification of Prox1, a homeobox transcription factor, as a key player in inhibiting neurite elongation. Although Prox1 promotes acquisition of early neuronal identity and is expressed in nascent post-mitotic neurons, it is heavily down-regulated in the majority of terminally differentiated neurons, indicating a regulatory role in delaying neurite outgrowth in newly formed neurons. Consistently, we show that Prox1 is sufficient to inhibit neurite extension in mouse and human neuroblastoma cell lines. More importantly, Prox1 overexpression suppresses neurite elongation in primary neuronal cultures as well as in the developing mouse brain, while Prox1 knock-down promotes neurite outgrowth. Mechanistically, RNA-Seq analysis reveals that Prox1 affects critical pathways for neuronal maturation and neurite extension. Interestingly, Prox1 strongly inhibits many components of Ca2+ signaling pathway, an important mediator of neurite extension and neuronal maturation. In accordance, Prox1 represses Ca2+ entry upon KCl-mediated depolarization and reduces CREB phosphorylation. These observations suggest that Prox1 acts as a potent suppressor of neurite outgrowth by inhibiting Ca2+ signaling pathway. This action may provide the appropriate time window for nascent neurons to find the correct position in the CNS prior to initiation of neurites and axon elongation.

Asunto(s)

Señalización del Calcio , Sistema Nervioso Central/patología , Proteínas de Homeodominio/metabolismo , Neuroblastoma/patología , Proyección Neuronal , Neuronas/patología , Proteínas Supresoras de Tumor/metabolismo , Animales , Células Cultivadas , Sistema Nervioso Central/metabolismo , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Proteínas de Homeodominio/genética , Humanos , Ratones , Neuroblastoma/metabolismo , Neuronas/metabolismo , Fosforilación , Transducción de Señal , Proteínas Supresoras de Tumor/genética

RESUMEN

A sensitive analytical method was developed and validated for the quantification of cotinine in mouse plasma after exposure to smoke of 0.5, 1.0, and 1.5 commercially available cigarettes, using liquid chromatography tandem mass spectrometry. The method was validated over a linear concentration range of 0.075-20.0 ng/mL with the R2 value being higher than 0.99. Both the precision (coefficient of variation; %) and accuracy (relative error; %) were within acceptable criteria of <15%. The lower limit of quantification (LLOQ) for cotinine was 0.075 ng/mL with sufficient specificity, accuracy, and precision. Following exposure to 0.5, 1.0, and 1.5 cigarette smoke, it was observed that the AUC and the Cmax increased linearly as the doses increased. The pharmacokinetics of cotinine was found linear for the range of 0.5-1.5 commercial cigarette smoke. The quantification of the concentration of cotinine in mouse plasma after smoke exposure will facilitate future behavioral and toxicological experiments in animals and may prove useful in predicting cotinine levels in humans during smoking.

Asunto(s)

Cotinina/sangre , Cotinina/farmacocinética , Exposición por Inhalación/análisis , Contaminación por Humo de Tabaco , Animales , Cotinina/química , Modelos Lineales , Masculino , Ratones , Ratones Endogámicos BALB C , Reproducibilidad de los Resultados , Sensibilidad y Especificidad

RESUMEN

The aim of the present study was to investigate the effect of rutin administration (100 mg/kg/day) to pentylenetetrazol (PTZ)-treated Balb-c mice (60 mg/kg/day), with respect to anxiety-like behavior using both open-field and elevated plus-maze (EPM) tests, and acetylcholinesterase (AChE) activity in salt-soluble (SS) fraction and detergent-soluble (DS) fraction of the cerebral cortex, hippocampus, striatum, midbrain, and diencephalon. Our results demonstrated that the administration of PTZ in 3 doses and the induction of seizures increased significantly anxiety behavior of mice and reduced significantly DS-AChE activity in all brain regions examined, while the reduction in the SS fraction was brain region-specific. Rutin administration to normal mice did not affect their behavior, while it induced a brain region-specific reduction in SS-AChE and a significant decrease in DS-AChE in all brain regions. We demonstrated for the first time that pretreatment of PTZ-mice with rutin (PTZ + Rutin group) prevented the manifestation of anxiety and induced interestingly a further significant reduction on the SS- and DS-AChE activities only in the cerebral cortex and striatum, in comparison with PTZ group. Our results show that rutin exhibits an important anxiolytic effect and an anticholinesterase activity in specific brain areas in the seizure model of PTZ.

Asunto(s)

Acetilcolinesterasa/metabolismo , Ansiedad/tratamiento farmacológico , Ansiedad/enzimología , Encéfalo/enzimología , Pentilenotetrazol/toxicidad , Rutina/uso terapéutico , Convulsiones/tratamiento farmacológico , Convulsiones/enzimología , Animales , Encéfalo/efectos de los fármacos , Isoenzimas/metabolismo , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Aprendizaje por Laberinto/fisiología , Ratones , Ratones Endogámicos BALB C , Rutina/farmacología , Convulsiones/inducido químicamente , Resultado del Tratamiento

RESUMEN

In the version of this Article originally published, in Fig. 1c-e, on the x axes, the lines labelled 'Aß42' and 'Aß42(F19S;L34P)' grouped the data incorrectly; the line labelled Aß42 should have grouped the data for Random 1-2 and Clones 1-10, and the line labelled Aß42(F19S;L34P) should have only grouped the data for Random 1-2 on the right end of the plots and blots. These figures have now been corrected in all versions of the Article.

RESUMEN

This study aimed to investigate the potential neurotoxic effects of aflatoxin B1 (AFB1) and the preventive effects of saffron. Male Balb-c mice received AFB1 (0.6 mg/kg/day intraperitoneally for 4 days), saffron infusion (90 mg styles/200 mL, ad libitum access for 2 weeks) or saffron infusion plus AFB1 (saffron treatment as previously plus 0.6 mg AFB1/kg/day intraperitoneally for the last 4 days). Control mice were intraperitoneally injected with DMSO:saline (1:1, v/v) during AFB1 treatment. Learning/memory was assessed by passive avoidance task. The activity of acetylcholinesterase [AChE, salt-(SS)/detergent-soluble(DS) isoforms], butyrylcholinesterase (BuChE, SS/DS isoforms), monoamine oxidase (MAO-A, MAO-B), the levels of lipid peroxidation (malondialdehyde, MDA) and reduced glutathione (GSH), were determined in whole brain (minus cerebellum) and cerebellum. We demonstrate for the first time that AFB1 administration impaired the memory of adult mice and decreased significantly whole brain AChE and BuChE activity, cerebellar AChE activity and cerebral GSH content. Moreover, MAO isoforms activity in whole brain, MAO-B activity in cerebellum and MDA levels of both tissues were significantly higher after AFB1 treatment. Pre-treatment with saffron prevented memory decline, activation of MAO-A and MAO-B in whole brain and cerebellum, respectively, and lipid peroxidation triggered by AFB1. Interestingly, the activity of AChE isoforms in whole brain, DS-AChE in cerebellum and GSH levels of both tissues were further significantly decreased in saffron +AFB1-treated mice compared with AFB1 group. Our findings support the neuroprotective efficacy of saffron against AFB1 in adult mice.

Asunto(s)

Encéfalo/efectos de los fármacos , Cognición/efectos de los fármacos , Aprendizaje/efectos de los fármacos , Memoria/efectos de los fármacos , Extractos Vegetales/farmacología , Aflatoxina B1/farmacología , Animales , Antioxidantes/farmacología , Encéfalo/metabolismo , Aprendizaje/fisiología , Peroxidación de Lípido/efectos de los fármacos , Masculino , Ratones Endogámicos BALB C , Estrés Oxidativo/efectos de los fármacos , Té

RESUMEN

A systematic data quality validation and normalization strategy is an important component of the omic profile meta-analysis, ensuring comparability of the profiles and exclusion of experimental biases from the derived biological conclusions. In this study, we present the normalization methodology applied on the sets of cerebellum gas chromatography-mass spectrometry metabolic profiles of 124days old male and female animals in an adult-onset-hypothyroidism (AOH) mouse model before combining them into a sex-comparative analysis. The employed AOH model concerns the monitoring of the brain physiology of Balb/cJ mice after eight-week administration of 1%w/v KClO4 in the drinking water, initiated on the 60th day of their life. While originating from the same animal study, the tissues of the two sexes were processed and their profiles acquired and analyzed at different time periods. Hence, the previously published profile set of male mice was first re-annotated based on the presently available resources. Then, after being validated as acquired under the same analytical conditions, both profiles sets were corrected for derivatization biases and filtered for low-confidence measurements based on the same criteria. The final normalized 73-metabolite profiles contribute to the currently few available omic datasets of the AOH effect on brain molecular physiology, especially with respect to sex differentiation. Multivariate statistical analysis indicated one (unknown) and three (succinate, benzoate, myristate) metabolites with significantly higher and lower, respectively, cerebellum concentration in the hypothyroid compared to the euthyroid female mice. The respective numbers for the males were two and 24. Comparison of the euthyroid cerebellum metabolic profiles between the two sexes indicated 36 metabolites, including glucose, myo- and scyllo-inositol, with significantly lower concentration in the females versus the males. This implies that the female mouse cerebellum has been conditioned to smaller changes in its metabolic activity with respect to the pathways involving these metabolites compared to the male animals. In conclusion, our study indicated a much subtler AOH effect on the cerebellum metabolic activity of the female compared to the male mice. The leaner metabolic profile of the female mouse cerebellum was suggested as a potential factor contributing to this phenomenon.

Asunto(s)

Cerebelo/metabolismo , Cromatografía de Gases y Espectrometría de Masas/métodos , Hipotiroidismo/metabolismo , Metabolómica/métodos , Animales , Peso Corporal , Cerebelo/química , Cerebelo/fisiopatología , Análisis por Conglomerados , Modelos Animales de Enfermedad , Femenino , Hipotiroidismo/fisiopatología , Masculino , Metaboloma , Ratones , Ratones Endogámicos BALB C , Análisis de Componente Principal , Factores Sexuales , Biología de Sistemas

RESUMEN

Protein misfolding and aggregation are common pathological features of several human diseases, including Alzheimer's disease and type 2 diabetes. Here, we report an integrated and generalizable bacterial system for the facile discovery of chemical rescuers of disease-associated protein misfolding. In this system, large combinatorial libraries of macrocyclic molecules are biosynthesized in Escherichia coli cells and simultaneously screened for their ability to rescue pathogenic protein misfolding and aggregation using a flow cytometric assay. We demonstrate the effectiveness of this approach by identifying drug-like, head-to-tail cyclic peptides that modulate the aggregation of the Alzheimer's disease-associated amyloid ß peptide. Biochemical, biophysical and biological assays using isolated amyloid ß peptide, primary neurons and various established Alzheimer's disease nematode models showed that the selected macrocycles potently inhibit the formation of neurotoxic amyloid ß peptide aggregates. We also applied the system to the identification of misfolding rescuers of mutant Cu/Zn superoxide dismutase-an enzyme linked with inherited forms of amyotrophic lateral sclerosis. Overall, the system enables the identification of molecules with therapeutic potential for rescuing the misfolding of disease-associated polypeptides.

RESUMEN

Metabolism being a fundamental part of molecular physiology, elucidating the structure and regulation of metabolic pathways is crucial for obtaining a comprehensive perspective of cellular function and understanding the underlying mechanisms of its dysfunction(s). Therefore, quantifying an accurate metabolic network activity map under various physiological conditions is among the major objectives of systems biology in the context of many biological applications. Especially for CNS, metabolic network activity analysis can substantially enhance our knowledge about the complex structure of the mammalian brain and the mechanisms of neurological disorders, leading to the design of effective therapeutic treatments. Metabolomics has emerged as the high-throughput quantitative analysis of the concentration profile of small molecular weight metabolites, which act as reactants and products in metabolic reactions and as regulatory molecules of proteins participating in many biological processes. Thus, the metabolic profile provides a metabolic activity fingerprint, through the simultaneous analysis of tens to hundreds of molecules of pathophysiological and pharmacological interest. The application of metabolomics is at its standardization phase in general, and the challenges for paving a standardized procedure are even more pronounced in brain studies. In this review, we support the value of metabolomics in brain research. Moreover, we demonstrate the challenges of designing and setting up a reliable brain metabolomic study, which, among other parameters, has to take into consideration the sex differentiation and the complexity of brain physiology manifested in its regional variation. We finally propose ways to overcome these challenges and design a study that produces reproducible and consistent results.

RESUMEN

Thyroid hormones (TH) are essential for normal development and function of mammalian central nervous system (CNS); TH dysregulation has been implicated in several cognitive and behavioral deficits related to dysfunctions of neurotransmitter systems. In the present study, we investigated the effects of adult onset hypothyroidism on the activity of acetylcholinesterase (AChE) and on related behavioral parameters. For this purpose we used adult male Balb/cJ mice that were divided randomly into euthyroid and hypothyroid animal groups. Animals were rendered hypothyroid through administration of 1% w/v KClO4 in their drinking water for 8weeks. At the end of the treatment, learning/memory procedures were examined through step-through passive avoidance task while fear/anxiety was assessed using elevated plus-maze (EPM) and open-field (OF) tests. AChE activity was determined colorimetrically in two different fractions, salt-soluble fraction (SS) (containing mainly the G1 isoform) and detergent-soluble fraction (DS) (containing mainly the G4 isoform) in cerebral cortex, cerebellum, midbrain, hippocampus and striatum. Our results indicate that adult onset hypothyroidism caused significant memory impairment and increased fear/anxiety. Moreover, the activity of both isoforms of AChE was reduced in all brain regions examined in a brain region- and isoform-specific manner.

Asunto(s)

Acetilcolinesterasa/metabolismo , Encéfalo/enzimología , Hipotiroidismo/enzimología , Hipotiroidismo/psicología , Edad de Inicio , Animales , Ansiedad/enzimología , Reacción de Prevención/fisiología , Modelos Animales de Enfermedad , Miedo/fisiología , Isoenzimas/metabolismo , Masculino , Trastornos de la Memoria/enzimología , Ratones Endogámicos BALB C , Actividad Motora/fisiología , Percloratos , Compuestos de Potasio , Distribución Aleatoria

RESUMEN

Our goal was to delineate the mechanisms of selenite-induced oxidative stress in neonatal rats and investigate the potential of blueberry leaf polyphenols to counteract the induced stress. Vaccinium corymbosum leaf decoction (BLD) was analyzed by UPLC-MS and LC-DAD, along with its in vitro antioxidant activity (DPPH radical scavenging, FRAP, ferrous chelation). Newborn suckling Wistar rats were randomly divided into three groups: 'Se' and 'SeBLD' received 20 µmol Na2SeO3/kg BW subcutaneously (PN day 10); 'SeBLD' received 100 mg dry BLD/kg BW intraperitoneally (PN11 and 12) and Group 'C' received normal saline. Βiochemical analysis revealed tissue-specific effects of selenite. Brain as a whole was more resistant to selenite toxicity in comparison to liver; midbrain and cerebellum were in general not affected, but cortex was moderately disturbed. Liver lipid peroxidation, GSH, SOD, CAT, GPx were significantly affected, whereas proteolytic activity was not. BLD, which is rich in chlorogenic acid and flavonols (especially quercetin derivatives), exerted significant antioxidant protective effects in all regions. In conclusion, we provide for the first time an insight to the neonatal rat cerebral and liver redox response against a toxic selenite dose and blueberry leaf polyphenols.

Asunto(s)

Antioxidantes/farmacología , Arándanos Azules (Planta)/química , Química Encefálica/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Polifenoles/farmacología , Ácido Selenioso/toxicidad , Animales , Animales Recién Nacidos , Antioxidantes/metabolismo , Femenino , Peroxidación de Lípido/efectos de los fármacos , Hígado/efectos de los fármacos , Masculino , Oxidación-Reducción , Extractos Vegetales/farmacología , Hojas de la Planta/química , Ratas , Ratas Wistar

RESUMEN

Evaluation of fear and anxiety levels offers valuable insight on the impact of experimental conditions. The elevated plus-maze and the open field (thigmotactic responce) tests are two well-established behavioral procedures for the quantification of anxiety in rodents. In this study, Phobos, a novel, effective and simple application developed for recording rodents' behavior during the elevated plus-maze and the open-field test, is being presented. Phobos is able to generate all basic locomotor-related behavioral results at once, immediately after a simple manual record of the rodent's position, along with simultaneous analysis of the experiment in 5-min periods. The efficiency of Phobos is demonstrated by presenting results from the two behavioral tests showing that animal's behavior unfolds differently in each one. Phobos manages to ease the experimenter from laborious work by providing self-explanatory characteristics and a convenient way to record the behavior of the animal, while it quickly calculates all basic locomotor-related parameters, easing behavioral studies. Phobos is freely accessible at https://sourceforge.net/projects/phobosapplication/.

Asunto(s)

Aprendizaje por Laberinto , Actividad Motora , Programas Informáticos , Animales , Masculino , Ratones Endogámicos BALB C

RESUMEN

Our aim was to investigate the possible effects of regular drinking of Rosmarinus officinalis L. leaf infusion on behavior and on AChE activity of mice. Rosemary tea (2% w/w) phytochemical profile was investigated through LC/DAD/ESI-MS(n). Adult male mice were randomly divided into two groups: "Rosemary-treated" that received orally the rosemary tea for 4weeks and "control" that received drinking water. The effects of regular drinking of rosemary tea on behavioral parameters were assessed by passive avoidance, elevated plus maze and forced swimming tests. Moreover, its effects on cerebral and liver cholinesterase (ChE) isoforms activity were examined colorimetricaly. Phytochemical analysis revealed the presence of diterpenes, flavonoids and hydroxycinnamic derivatives in rosemary tea; the major compounds were quantitatively determined. Its consumption rigorously affected anxiety/fear and depression-like behavior of mice, though memory/learning was unaffected. ChE isoforms activity was significantly decreased in brain and liver of "rosemary treated" mice. In order to explain the tissue ChE inhibition, principal component analysis, pharmacophore alignment and molecular docking were used to explore a possible relationship between main identified compounds of rosemary tea, i.e. rosmarinic acid, luteolin-7-O-glucuronide, caffeic acid and known AChE inhibitors. Results revealed potential common pharmacophores of the phenolic components with the inhibitors. Our findings suggest that rosemary tea administration exerts anxiolytic and antidepressant effects on mice and inhibits ChE activity; its main phytochemicals may function in a similar way as inhibitors.

Asunto(s)

Ansiedad/prevención & control , Encéfalo/enzimología , Inhibidores de la Colinesterasa/farmacología , Colinesterasas/metabolismo , Depresión/prevención & control , Hígado/enzimología , Rosmarinus/química , Té , Animales , Simulación por Computador , Masculino , Espectrometría de Masas , Ratones , Ratones Endogámicos BALB C

RESUMEN

The aim of the present study was to investigate whether the underlying mechanism of lead (Pb)-induced effects on learning/memory and fear/anxiety behavior involves changes either on AChE G4 (most abundant in brain) or on G1 isoform activity, and/or to a putative local disruption of oxidant/antioxidant balance. Adult male mice were randomly divided into two groups (18 animals/group): a vehicle group [500ppm (mg/L) CH3COONa/day for 4weeks in their drinking water] and a Pb-treated group [500ppm Pb(CH3COO)2/day for 4weeks in their drinking water]. At the end of the treatment period, mice were subjected to the behavioral tasks. Learning/memory was tested by step-through passive avoidance test, whereas fear/anxiety was studied using the elevated plus-maze and thigmotaxis tests. Pb levels in mice brain were determined using atomic absorption spectrometry. AChE activity was determined colorimetrically, and GSH and MDA levels fluorometrically in whole brain minus cerebellum, cerebral cortex, midbrain, hippocampus, striatum and cerebellum. The possible correlations between learning/memory or fear/anxiety behavior with the AChE activity and/or the lipid peroxidation levels and GSH content were also examined. Pb consumption caused significant deficits on mice learning/memory ability and increased anxiety. The consumption of the Pb solution inhibited the activity of the two AChE isoforms in all brain regions tested. Moreover, Pb exposure increased lipid peroxidation and decreased GSH levels in all brain regions examined. Spearman correlation analysis revealed that the coefficients between the particular behaviors, AChE activity and redox balance were brain region- and AChE isoform-specific.

Asunto(s)

Acetilcolinesterasa/metabolismo , Trastornos de Ansiedad/fisiopatología , Encéfalo/fisiopatología , Intoxicación del Sistema Nervioso por Plomo en Adultos/fisiopatología , Trastornos de la Memoria/fisiopatología , Animales , Trastornos de Ansiedad/etiología , Miedo/fisiología , Glutatión/metabolismo , Isoenzimas/metabolismo , Intoxicación del Sistema Nervioso por Plomo en Adultos/complicaciones , Aprendizaje/fisiología , Peroxidación de Lípido/fisiología , Masculino , Malondialdehído/metabolismo , Trastornos de la Memoria/etiología , Ratones Endogámicos BALB C , Pruebas Neuropsicológicas

RESUMEN

PURPOSE: The goals of this study were to monitor the effect of drinking of herbal tea from Sideritis clandestina subsp. clandestina for 6 weeks on behavioral and oxidant/antioxidant parameters of adult male mice and also to evaluate its phytochemical composition. METHODS: The phytochemical profile of the Sideritis tea was determined by liquid chromatography-UV diode array coupled to ion-trap mass spectrometry with electrospray ionization interface. The effects of two doses of the herbal infusion (2 and 4% w/v, daily) intake on anxiety-like state in mice were studied by the assessment of their thigmotactic behavior. The oxidant/antioxidant status of brain (-Ce), liver and heart of adult male Balb-c mice following the consumption of Sideritis tea was also evaluated via the measurement of malondialdehyde (MDA) and reduced glutathione (GSH) levels using fluorometric assays. Our study was further extended to determine the antioxidant effects of the herbal tea on specific brain regions (cerebral cortex, cerebellum and midbrain). RESULTS: The identified compounds were classified into several natural product classes: quinic acid derivatives, iridoids, phenylethanol glycosides and flavonoids. Our results showed that only the 4% Sideritis tea exhibited anxiolytic-like properties as evidenced by statistically significant (p < 0.05) decrease in the thigmotaxis time and increase in the number of entries to the central zone in comparison with the control group. Consumption of both tea doses (2 and 4% w/v) elevated GSH (12 and 28%, respectively, p < 0.05) and decreased MDA (16 and 29%, p < 0.05) levels in brain (-Ce), while liver and heart remained unaffected. In regard to the effect of herbal tea drinking (2 and 4% w/v) on specific brain regions, it caused a significant increase in GSH of cerebellum (13 and 36%, respectively, p < 0.05) and midbrain (17 and 36%, p < 0.05). Similarly, MDA levels were decreased in cerebellum (45 and 79%, respectively, p < 0.05) and midbrain (50 and 63%, respectively, p < 0.05), whereas cerebral cortex remained unaffected. CONCLUSIONS: Mountain tea drinking prevents anxiety-related behaviors and confers antioxidant protection to rodent's tissues in a region-specific, dose-dependent manner, and its phytochemical constituents are shown for the first time.

Asunto(s)

Antioxidantes/farmacología , Bebidas , Extractos Vegetales/farmacología , Sideritis/química , Animales , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Cromatografía Liquida , Flavonoides/farmacología , Glutatión/análisis , Glicósidos/farmacología , Corazón/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Malondialdehído/análisis , Espectrometría de Masas , Ratones , Ratones Endogámicos BALB C

RESUMEN

In the present study, the possible reversal effects of saffron against established aluminum (Al)-toxicity in adult mice, were investigated. Control, Al-treated (50 mg AlCl(3)/kg/day diluted in the drinking water for 5 weeks) and Al+saffron (Al-treatment as previously plus 60 mg saffron extract/kg/day intraperitoneally for the last 6 days), groups of male Balb-c mice were used. We assessed learning/memory, the activity of acetylcholinesterase [AChE, salt-(SS)/detergent-soluble(DS) isoforms], butyrylcholinesterase (BuChE, SS/DS isoforms), monoamine oxidase (MAO-A, MAO-B), the levels of lipid peroxidation (MDA) and reduced glutathione (GSH), in whole brain and cerebellum. Brain Al was determined by atomic absorption spectrometry, while, for the first time, crocetin, the main active metabolite of saffron, was determined in brain after intraperitoneal saffron administration by HPLC. Al intake caused memory impairment, significant decrease of AChE and BuChE activity, activation of brain MAO isoforms but inhibition of cerebellar MAO-B, significant elevation of brain MDA and significant reduction of GSH content. Although saffron extract co-administration had no effect on cognitive performance of mice, it reversed significantly the Al-induced changes in MAO activity and the levels of MDA and GSH. AChE activity was further significantly decreased in cerebral tissues of Al+saffron group. The biochemical changes support the neuroprotective potential of saffron under toxicity.

Asunto(s)

Aluminio/toxicidad , Encéfalo/efectos de los fármacos , Crocus/química , Memoria/efectos de los fármacos , Fármacos Neuroprotectores/farmacología , Acetilcolinesterasa/metabolismo , Aluminio/análisis , Aluminio/farmacocinética , Animales , Conducta Animal/efectos de los fármacos , Encéfalo/metabolismo , Carotenoides/análisis , Carotenoides/metabolismo , Cognición/efectos de los fármacos , Glutatión/metabolismo , Inyecciones Intraperitoneales , Aprendizaje/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/enzimología , Malondialdehído/metabolismo , Trastornos de la Memoria/inducido químicamente , Trastornos de la Memoria/tratamiento farmacológico , Ratones , Ratones Endogámicos BALB C , Monoaminooxidasa/metabolismo , Estrés Oxidativo , Extractos Vegetales/farmacología , Vitamina A/análogos & derivados

RESUMEN

Inhibitors of acetylcholine breakdown by acetylcholinesterase (AChE) constitute the main therapeutic modality for Alzheimer's disease. In the search for natural products with inhibitory action on AChE, this study investigated the activity of saffron extract and its constituents by in vitro enzymatic and molecular docking studies. Saffron has been used in traditional medicine against Alzheimer's disease. Saffron extract showed moderate AChE inhibitory activity (up to 30%), but IC(50) values of crocetin, dimethylcrocetin, and safranal were 96.33, 107.1, and 21.09 µM, respectively. Kinetic analysis showed mixed-type inhibition, which was verified by in silico docking studies. Safranal interacts only with the binding site of the AChE, but crocetin and dimethylcrocetin bind simultaneously to the catalytic and peripheral anionic sites. These results reinforce previous findings about the beneficial action of saffron against Alzheimer's disease and may be of value for the development of novel therapeutic agents based on carotenoid-based dual binding inhibitors.

Asunto(s)

Inhibidores de la Colinesterasa , Crocus/química , Acetilcolinesterasa/metabolismo , Enfermedad de Alzheimer/tratamiento farmacológico , Sitios de Unión , Carotenoides/química , Carotenoides/metabolismo , Inhibidores de la Colinesterasa/metabolismo , Inhibidores de la Colinesterasa/farmacología , Inhibidores de la Colinesterasa/uso terapéutico , Humanos , Cinética , Modelos Moleculares , Extractos Vegetales/farmacología , Relación Estructura-Actividad , Vitamina A/análogos & derivados

RESUMEN

Amyloid precursor protein (APP) altered metabolism, Aß-overproduction/aggregation and oxidative stress are implicated in the development of Alzheimer's disease pathology. Based on our previous data indicating that administration of a polyphenol-rich (PrB) blueberry extract (from wild Vaccinium angustifolium) is memory enhancing in healthy mice and in order to delineate the neuroprotective mechanisms, this study investigated the antioxidant effects of PrB in H2O2-induced oxidative damage, Aß peptide fibrillogenesis and APP metabolism. PrB suppressed H2O2-initiated oxidation (DCF assay) and cell death (MTT assay) in SH-SY5Y cells. Protective effects were observed on Chinese hamster ovary (CHO) cells overexpressing APP770 carrying the mutation Val717Phe only at high concentrations, while further damage on HEK293 cells was induced after co-treatment with 250 µM H2O2 and PrB in comparison with H2O2 alone. Using the thioflavine T assay, blueberry polyphenols inhibited Aß-aggregation (~70%, 15 µg/mL) in a time-dependent manner, while in the CHO(APP770) cells it had no effect on APP metabolism as assessed by western blot. The results suggest that blueberry polyphenols exhibit antioxidant and/or pro-oxidant properties according to the cellular environment and have no effect on APP metabolism.

Asunto(s)

Precursor de Proteína beta-Amiloide/metabolismo , Arándanos Azules (Planta)/química , Peróxido de Hidrógeno/toxicidad , Polifenoles/farmacología , Animales , Antocianinas/farmacología , Células CHO , Supervivencia Celular , Cricetinae , Frutas/química , Células HEK293 , Humanos , Estrés Oxidativo , Extractos Vegetales/farmacología , Especies Reactivas de Oxígeno/análisis

RESUMEN

Oxidative stress is involved in the pathophysiology of neurodegenerative diseases and aging. Many species of the genus Sideritis (mountain tea) are widely consumed in the Mediterranean region as herbal tea. This study evaluated the effect of supplementation of mice with herbal tea from Sideritis clandestina subsp. peloponnesiaca on the antioxidant status of different brain regions. To select the most bioactive herbal tea, the polyphenolic content (Folin-Ciocalteu method) and the antioxidant properties (ferric reducing antioxidant power [FRAP] and 2,2-diphenyl-1-picrylhydrazyl assays) of several taxa and different populations of the S. clandestina infusions were measured in vitro. Male adult mice had ad libitum access to water (control) or the herbal tea (4% w/v) for 6 weeks. At the end of the treatment period we assessed the total antioxidant power (FRAP assay) and the levels of malondialdehyde (indicator of lipid peroxidation) and reduced glutathione in the cerebral cortex, cerebellum, and midbrain. These biochemical measures have also been determined in liver samples used as a comparative reference peripheral tissue. Consumption of 4% herbal tea increased the total antioxidant power of the midbrain by 72% (P<.05); a significant (P<.05) decrease in malondialdehyde levels and increase in reduced glutathione content of the cerebellum (78% and 27%, respectively) and midbrain (59% and 32%, respectively) were also observed. These findings indicate that mountain tea consumption enhances the antioxidant defense of the adult rodent brain in a region-specific manner.

Asunto(s)

Antioxidantes/metabolismo , Bebidas , Cerebelo/metabolismo , Corteza Cerebral/metabolismo , Mesencéfalo/metabolismo , Sideritis/química , Animales , Flores/química , Glutatión/metabolismo , Peroxidación de Lípido , Masculino , Malondialdehído/metabolismo , Ratones , Ratones Endogámicos BALB C , Oxidación-Reducción , Hojas de la Planta/química , Tallos de la Planta/química , Polifenoles/análisis , Especificidad de la Especie

RESUMEN

Brain aging is characterized by cognitive decline and memory deficits that could be the result of oxidative stress and impaired cholinergic function. In this study, the effects of a daily, 7-day, intraperitoneal administration of saffron on cognitive functions were examined in both healthy adult (4 months old) and aged (20 months old), male Balb-c mice (n=8/group), by passive avoidance test. Whole brain homogenates (minus cerebellum) were collected for examination of brain oxidative markers, caspase-3 and acetylcholinesterase (AChE) activity. Results showed that saffron-treated mice exhibited significant improvement in learning and memory, accompanied by reduced lipid peroxidation products, higher total brain antioxidant activity and reduced caspase-3 activity in both age groups of mice. Furthermore, salt- and detergent-soluble AChE activity was significantly decreased only in adult mice. Thus, we showed, for the first time, that the significant cognitive enhancement conferred by saffron administration in mice, is more closely related to the antioxidant reinforcement. Next, we compared the effect of saffron (1-250 µg/mL), crocetin and safranal (1-125 µM) on H(2)O(2)-induced toxicity in human neuroblastoma SH-SY5Y cells. Both saffron and crocetin provided strong protection in rescuing cell viability (MTT assay), repressing ROS production (DCF assay) and decreasing caspase-3 activation. These data, together with earlier studies suggest that crocetin is a unique and potent antioxidant, capable of mediating the in vivo effects of saffron.

Asunto(s)

Envejecimiento/psicología , Antioxidantes/farmacología , Crocus , Memoria/efectos de los fármacos , Acetilcolinesterasa/metabolismo , Animales , Reacción de Prevención/efectos de los fármacos , Química Encefálica/efectos de los fármacos , Carotenoides/farmacología , Caspasa 3/metabolismo , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Humanos , Peróxido de Hidrógeno/toxicidad , Inyecciones Intraperitoneales , Aprendizaje/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos BALB C , Oxidantes/toxicidad , Extractos Vegetales/administración & dosificación , Extractos Vegetales/farmacología , Especies Reactivas de Oxígeno/metabolismo , Vitamina A/análogos & derivados

RESUMEN

Although adult-onset hypothyroidism (AOH) has been connected to neural activity alterations, including movement, behavioral, and mental dysfunctions, the underlying changes in brain metabolic physiology have not been investigated in a systemic and systematic way. The current knowledge remains fragmented, referring to different experimental setups and recovered from various brain regions. In this study, we developed and applied a gas chromatography-mass spectrometry (GC-MS) metabolomics protocol to obtain a holistic view of the cerebellar metabolic physiology in a Balb/cJ mouse model of prolonged adult-onset hypothyroidism induced by a 64-day treatment with 1% potassium perchlorate in the drinking water of the animals. The high-throughput analysis enabled the correlation between multiple parallel-occurring metabolic phenomena; some have been previously related to AOH, while others implicated new pathways, designating new directions for further research. Specifically, an overall decline in the metabolic activity of the hypothyroid compared to the euthyroid cerebellum was observed, characteristically manifested in energy metabolism, glutamate/glutamine metabolism, osmolytic/antioxidant capacity, and protein/lipid synthesis. These alterations provide strong evidence that the mammalian cerebellum is metabolically responsive to AOH. In light of the cerebellum core functions and its increasingly recognized role in neurocognition, these findings further support the known phenotypic manifestations of AOH into movement and cognitive dysfunctions.

Asunto(s)

Cerebelo/metabolismo , Cromatografía de Gases y Espectrometría de Masas/métodos , Hipotiroidismo/metabolismo , Metabolómica/métodos , Animales , Peso Corporal , Cerebelo/química , Análisis por Conglomerados , Modelos Animales de Enfermedad , Hipotiroidismo/inducido químicamente , Masculino , Redes y Vías Metabólicas , Metaboloma , Ratones , Ratones Endogámicos BALB C , Análisis por Micromatrices , Percloratos , Compuestos de Potasio , Análisis de Componente Principal , Reproducibilidad de los Resultados