RESUMO
Aversive stimuli activate corticotropin-releasing factor (CRF)-expressing neurons in the paraventricular nucleus of hypothalamus (PVNCRF neurons) and other brain stress systems to facilitate avoidance behaviors. Appetitive stimuli also engage the brain stress systems, but their contributions to reward-related behaviors are less well understood. Here, we show that mice work vigorously to optically activate PVNCRF neurons in an operant chamber, indicating a reinforcing nature of these neurons. The reinforcing property of these neurons is not mediated by activation of the hypothalamic-pituitary-adrenal (HPA) axis. We found that PVNCRF neurons send direct projections to the ventral tegmental area (VTA), and selective activation of these projections induced robust self-stimulation behaviors, without activation of the HPA axis. Similar to the PVNCRF cell bodies, self-stimulation of PVNCRF-VTA projection was dramatically attenuated by systemic pretreatment of CRF receptor 1 or dopamine D1 receptor (D1R) antagonist and augmented by corticosterone synthesis inhibitor metyrapone, but not altered by dopamine D2 receptor (D2R) antagonist. Furthermore, we found that activation of PVNCRF-VTA projections increased c-Fos expression in the VTA dopamine neurons and rapidly triggered dopamine release in the nucleus accumbens (NAc), and microinfusion of D1R or D2R antagonist into the NAc decreased the self-stimulation of these projections. Together, our findings reveal an unappreciated role of PVNCRF neurons and their VTA projections in driving reward-related behaviors, independent of their core neuroendocrine functions. As activation of PVNCRF neurons is the final common path for many stress systems, our study suggests a novel mechanism underlying the positive reinforcing effect of stressful stimuli.
Assuntos
Hormônio Liberador da Corticotropina , Hormônios Liberadores de Hormônios Hipofisários , Camundongos , Animais , Hormônio Liberador da Corticotropina/metabolismo , Hormônios Liberadores de Hormônios Hipofisários/metabolismo , Hormônios Liberadores de Hormônios Hipofisários/farmacologia , Sistema Hipotálamo-Hipofisário , Sistema Hipófise-Suprarrenal , Hipotálamo/metabolismo , Núcleo Hipotalâmico Paraventricular/metabolismo , Neurônios Dopaminérgicos/metabolismoRESUMO
Heating edible oils generates aldehydes, potentially leading to adverse health effects, making their analysis essential for quality control. This study presents a convenient miniaturized kapok fiber-supported liquid-phase extraction/in-situ derivatization method for the simultaneous extraction and derivatization of aldehydes in oils. The method involves placing 150 mg oil into a 1 mL pipette tip packed with 25 mg kapok fiber, adding 150 µL ACN with 1.5 mg mL-1 DNPH, and post 30-minute static extraction, retrieving the extractant with a pipettor for liquid chromatography-tandem mass spectrometry analysis. By optimizing critical parameters through a Box-Behnken design, the method exhibits good linearity (1-500 ng g-1, R2 ≥ 0.991), low detection limits (0.2-1.0 ng g-1), excellent accuracy (95.3-107.1%) and high precisions (relative standard deviation < 7.9%). This method simplifies sample preparation processes, cuts solvent use, and facilitates automation. It effectively identifies ten aldehyde variations in six heated oils, displaying distinct profiles consistent with prior research.
Assuntos
Aldeídos , Espectrometria de Massas em Tandem , Espectrometria de Massas em Tandem/métodos , Aldeídos/análise , Cromatografia Líquida , Extração Líquido-Líquido/métodos , Óleos de Plantas/química , Cromatografia Líquida de Alta Pressão/métodosRESUMO
The current study proposed a novel feather fiber-supported liquid extraction (FF-SLE) method for extracting analytes from oil samples. The natural feather fibers were used as the oil support material and directly loaded in the plastic tube of a disposable syringe to construct the low-cost extraction device (â¼0.5 CNY). The edible oil without any pretreatment including dilution was added directly to the extraction device, followed by the addition of the green extraction solvent of ethanol. As an example, the proposed method was applied to extract nine synthetic antioxidants from edible oils. The optimized extraction conditions for processing 0.5 g of oil were obtained when the syringe dimension was 5 mL, the extraction solvent was 0.5 mL of ethanol, the amount of feather fibers was 200 mg of duck feather fibers and the static extraction time was 10 min. The applications to seven kinds of feathers and seven kinds of edible oils all indicated the excellent oil removal efficiencies (>98.0%). Combined with high-performance liquid chromatography-ultraviolet, a quantification method was validated with satisfied linearity (R2≥0.994), accuracy (95.8-114.6%) and precision (≤8.3%) with the limits of detection ranging from 50 to 100 ng/g. The proposed FF-SLE method was simple, effective, convenient, low-cost, green and environmental-friendly for the extraction of analytes from oil samples prior to instrument analysis.
Assuntos
Antioxidantes , Animais , Antioxidantes/análise , Cromatografia Líquida de Alta Pressão/métodos , Etanol , Plumas/química , Óleos de Plantas/análise , SolventesRESUMO
In this paper, a miniaturized kapok fiber-supported liquid extraction (mini-KF-SLE) method was proposed for selective extraction of pesticide residues in vegetable oils. The natural kapok fiber was used as an inert oil support material based on its hydrophobic and lipophilic properties, and the extraction device was conveniently constructed by loading 15 mg of kapok fiber at the lower middle part of a 1-mL pipette tip. The vegetable oil sample (150 mg) without any pretreatment was directly loaded, followed by the addition of 150 µL of acetonitrile (ACN) as the extractant. After static extraction for 30 min, the extractant was pipetted out with a pipettor. As the proof of concept, it was applied for extracting eight organochlorine pesticides (OCPs) from vegetable oils and the eluate was analyzed by gas chromatography-electron capture detector (GC-ECD). Under optimized conditions, the extraction recoveries of OCPs were calculated to be in ranges of 35.8-79.5%. The satisfied quantitation ability was verified by the established method with coefficients of determination (R2) being greater than 0.99. The limits of detection (LODs) were in ranges of 2.0-50.0 ng/g. The relative recoveries were in ranges of 78.3-117.0% with the inter-/intra-day relative standard deviation (RSD) both being less than 13.3%. The potential of mini-KF-SLE to extract other kinds of pesticides was further verified by the successful extracting three triazole pesticides in vegetable oils with good extraction recoveries (>41.4%). The proposed mini-KF-SLE in combination with instrument detection techniques has the great potential in the low-cost and high-throughput determination of various pesticide residues in vegetable oils.
Assuntos
Resíduos de Praguicidas , Óleos de PlantasRESUMO
In the current study, natural cotton fiber was served as the supporter of water, and the water acted as an extractant for liquid-phase microextraction of polar components in low-polar edible oils. An in-syringe extraction device was constructed to facilitate the extraction process by simply loading a certain amount of cotton fibers between the syringe needle and the plastic syringe tube. Then, the extraction process can be conveniently conducted by pull-push the syringe plunger. It can be regarded as a new type of dynamic liquid-phase microextraction method while operated more convent. For the feasibility study, the novel in-syringe cotton fiber-supported liquid extraction (CF-SLECF-SLE) pretreatment method was applied to extract free 3-mono-chloropropane-1,2-diol (3-MCPD) in edible oils. Specifically, the cotton fibers supported a certain amount of water by successfully pulling-pushing 1 mL of water and 1 mL of HEX in/out twice, respectively. Then, 2.0 mL of diluted oil sample (containing 0.4 g oil) was loaded in and out four times for extraction, during which process 3-MCPD was extracted into the supported water. The extracted 3-MCPD was desorbed with 1 mL of ethyl acetate (EA), derivatized with trimethyl silane imidazole (TMSI), and analyzed by gas chromatography-tandem mass spectrometry (GC-MS/MS). For three different spiked edible oils, the internal standard normalized matrix effect (IS-normalized ME) values were in ranges of 96.3-104.8% with RSD being 4.3%, benefiting the accurate quantitative analysis. The limit of quantification (LOQ) was calculated to be 2 ng/g, which met the regular determination requirement of 3-MCPD in edible oils. Satisfied linearity was obtained in 2-500 ng/g, with correlation coefficients (R2) being 0.998. The relative recoveries were in the ranges of 96.9-110.5%. The intra-/inter-day RSDs were less than 8.2% and 10.2%, respectively. The proposed method provides an efficient, simple, low-cost, and easy to automate strategy for determining free 3-MCPD in edible oils.
Assuntos
alfa-Cloridrina , Fibra de Algodão , Cromatografia Gasosa-Espectrometria de Massas/métodos , Óleos de Plantas/química , Seringas , Espectrometria de Massas em Tandem/métodos , Água , alfa-Cloridrina/análiseRESUMO
Resveratrol (RSV) is reported to have renoprotective activity against diabetic nephropathy, while the mechanisms underlying its function have not been fully elucidated. In this study, we investigate the effect and related mechanism of RSV against high glucose-induced epithelial to mesenchymal transition (EMT) in human tubular epithelial cells (HK-2). A typical EMT is induced by high glucose in HK-2 cells, accompanied by increased levels of reactive oxygen species (ROS). RSV exhibits a strong ability to inhibit high glucose-induced EMT by decreasing intracellular ROS levels via down-regulation of NADPH oxidase subunits NOX1 and NOX4. The activation of extracellular signal-regulated kinase (ERK1/2) is found to be involved in high glucose-induced EMT in HK-2 cells. RSV, like NADPH oxidase inhibitor diphenyleneiodonium, can block ERK1/2 activation induced by high glucose. Our results demonstrate that RSV is a potent agent against high glucose-induced EMT in renal tubular cells via inhibition of NADPH oxidase/ROS/ERK1/2 pathway.