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Oil adulteration is a global challenge in the production of high value-added natural oils. Raman spectroscopy combined with mathematical modeling can be used for adulteration detection of camellia oil (CAO). In this study, the advantages of traditional chemometrics and deep learning methods in identifying and quantifying adulterated CAO were compared from a statistical perspective, and no significant difference were founded in the identification of CAO at different levels of adulteration. The recognition rate of pure and adulterated CAO was 100 %, but there were misclassifications among different adulterated CAOs. The deep learning models outperformed chemometrics methods in quantitative prediction of adulteration level, with RP2, RMSEP, and RPD of the optimal ConvLSTM model achieved 0.999, 0.9 % and 31.5, respectively. The classifiers and models developed in this study based on deep learning have wide applicability and reliability, and provide a fast and accurate method for adulteration detection in CAO.
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Microplastics released from plastic-based filter bags during tea brewing have attracted widespread attention. Laser confocal micro-Raman and direct classical least squares were used to identify and estimate micron-sized microplastics. Characteristic peaks from pyrolysis-gas chromatography/mass spectrometry of polyethylene terephthalate, polypropylene, and nylon 6 were selected to construct curves for quantification submicron-sized microplastics. The results showed that microplastics released from tea bags in the tea infusions ranged from 80 to 1288 pieces (micron-sized) and 0 to 63.755 µg (submicron-sized) per filter bag. Nylon 6 woven tea bags released far fewer microplastics than nonwoven filter bags. In particular, a simple strategy of three pre-washes with room temperature water significantly reduced microplastic residues with removal rates of 76 %-94 % (micron-sized) and 80 %-87 % (submicron-sized), respectively. The developed assay can be used for the quantitative evaluation of microplastics in tea infusions, and the pre-washing reduced the risk of human exposure to microplastics during tea consumption.
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Caprolactama/análogos & derivados , Microplásticos , Poluentes Químicos da Água , Humanos , Plásticos/análise , Polímeros , Chá , Poluentes Químicos da Água/análise , Monitoramento Ambiental/métodosRESUMO
BACKGROUND: Selenium is an important nutritional supplement that mainly exists naturally in soil as inorganic selenium. Saccharomyces cerevisiae cells are excellent medium for converting inorganic selenium in nature into organic selenium. RESULTS: Under the co-stimulation of sodium selenite (Na2SeO3) and potassium selenite (K2SeO3), the activity of selenophosphate synthetase (SPS) was improved up to about five folds more than conventional Na2SeO3 group with the total selenite salts content of 30 mg/L. Transcriptome analysis first revealed that due to the sharing pathway between sodium ion (Na+) and potassium ion (K+), the K+ largely regulates the metabolisms of amino acid and glutathione under the accumulation of selenite salt. Furthermore, K+ could improve the tolerance performance and selenium-biotransformation yields of Saccharomyces cerevisiae cells under Na2SeO3 salt stimulation. CONCLUSION: The important role of K+ in regulating the intracellular selenium accumulation especially in terms of amino acid metabolism and glutathione, suggested a new direction for the development of selenium-enrichment supplements with Saccharomyces cerevisiae cell factory. © 2024 Society of Chemical Industry.
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Saccharomyces , Selênio , Selênio/metabolismo , Saccharomyces/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Selenito de Sódio/metabolismo , Ácido Selenioso/metabolismo , Glutationa/metabolismo , Sódio/metabolismo , Aminoácidos/metabolismo , Potássio/metabolismoRESUMO
The health risks posed by harmful substances resulting from the thermal degradation of frying oils are of great concern. Characteristic peak intensity ratios (PIRs) screened from Raman spectra were used to characterize the thermal degradation. High correlation coefficients between PIRs and acid values (AVs) of 0.972 (linear fitting), 0.984 (logarithmic function fitting), and 0.954 (linear fitting) for fried soybean oil, canola oil, and palm oil, were obtained at the PIRs of I1267/I1749, I1267/I1659, and I1267/I1749, respectively. The highly correlated PIRs common to the three oils were determined by Pearson's correlation coefficient combined with heat maps. To accommodate both linear and nonlinear features, a global model for predicting AVs of multi-varieties frying oils was constructed using a least-squares support vector machine algorithm, and the results performed well with a root mean square error of prediction of 0.016 and a ratio of prediction to deviation of 11.351. The whole results demonstrate that Raman spectroscopy could characterize the thermal degradation and has excellent quantitative analysis ability for food control based on AV in frying oils, thus providing a new approach to quality control of frying oils.
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Óleos , Análise Espectral Raman , Óleo de Brassica napus , Óleo de Palmeira , Ácidos , Óleos de Plantas/químicaRESUMO
Pesticides residues in foodstuffs are longstanding of great concern to consumers and governments, thus reliable evaluation techniques for these residues are necessary to ensure food safety. Emerging ambient ionization mass spectrometry (AIMS), a transformative technology in the field of analytical chemistry, is becoming a promising and solid evaluation technology due to its advantages of direct, real-time and in-situ ionization on samples without complex pretreatments. To provide useful guidance on the evaluation techniques in the field of food safety, we offered a comprehensive review on the AIMS technology and introduced their novel applications for the analysis of residual pesticides in foodstuffs under different testing scenarios (i.e., quantitative, screening, imaging, high-throughput detection and rapid on-site analysis). Meanwhile, the creative combination of AIMS with high-resolution mass analyzer (e.g., orbitrap and time-of-flight) was fundamentally mentioned based on recent studies about the detection and evaluation of multi-residual pesticides between 2015 and 2021. Finally, the technical challenges and prospects associated with AIMS operation in food industry were discussed.
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Resíduos de Praguicidas , Praguicidas , Praguicidas/análise , Radar , Espectrometria de Massas/métodos , Resíduos de Praguicidas/análise , Inocuidade dos AlimentosRESUMO
Widespread use of organophosphorus pesticides (OPs), especially superfluous and unreasonable use, had brought huge harm to the environment and food chain. It is because only a small part of the pesticides sprayed reached the target, and the rest slid across the soil, causing pollution of groundwater and surface water resources. These pesticides accumulate in the environment, causing environmental pollution. Therefore, in recent years, the control and degradation of OPs have become a public spotlight and research hotspot. Due to its unique advantages such as versatility, environmental compatibility, controllability, and cost-effectiveness compatibility, electrochemical technology has become one of the most promising methods for degradation of OPs. The fundamental knowledge about electrochemical degradation on OPs was introduced in this review. Then, a comprehensive overview of four main types of practical electrochemical technologies to degrade pesticides were presented and evaluated. The knowledge contained herein should conduce to better understand the degradation of pesticides by electrochemical technology, and better exploit the degradation of pesticides in the environment and food. Overall, the objective of this review is to provide comprehensive guidance for rational design and application of electrochemical technology in the degradation of OPs for the safety of the environment and food chain in the future.
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Praguicidas , Compostos Organofosforados/metabolismo , TecnologiaRESUMO
Microplastic (MP) contamination is a public issue for the environment and for human health. Plastic-based food filter bags, including polyethylene terephthalate, polypropylene, nylon 6 (NY6), and polyethylene, are widely used for soft drink sub-packaging, increasing the risk of MPs in foods and the environment. Three types of commercially available filter bags, including non-woven and woven bags, were collected, and MPs released after soaking were mapped using Raman imaging combined with chemometrics. Compared with peak area imaging at a single characteristic peak, Raman imaging combined with direct classical least squares calculation was more efficient and reliable for identifying MP features. Up to 94% of the bags released MPs after soaking, and there was no significant correlation with soaking conditions. Most MPs were tiny fragments and particles, and a few were fibrous MPs 620-840 µm in size. Woven NY6 filter bags had the lowest risk of releasing MPs. Source exploration revealed that most MPs originated from fragments and particles adsorbed on the surface of bags and strings. The results of this study are applicable to filter bag risk assessment and provide scientific guidance for regulating MPs in food.
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Cerebral stroke, a common clinical problem, is the predominant cause of disability and death worldwide. Its prevalence increases and infarctions exacerbate with age. A Tibetan plant, Brassica rapa L., possesses multiple medicinal effects, such as anti-altitude sickness, anti-hyperlipidemia and anti-fatigue, as mentioned in the noted ancient Tibet pharmacopeia "The Four Medical Tantras". Our preliminary studies also showed the anti-hypoxia protection mechanism of B. rapa L., implying its possible relationship with anti-ischemic neuroprotection. However, the potential molecular mechanism of the active constituent of turnip against cerebral ischemia/reperfusion remains unclear. In our study, oxidative stress markers, including LDH, ROS, SOD, GPx and CAT were assayed. In controlled in vitro assays, we found that the turnip's active constituent had remarkable anti-hypoxia capability. We further showed the profound effects of the active constituent of turnip on the levels of apoptosis-related proteins, including Bax, Bcl-2 and caspase-3, which contributed to its anti-inflammatory activity. Western blot analysis results also implied that active-constituent pretreatment reversed the diminished expression of the PI3K/Akt/mTOR pathway mediated by oxygen glucose deprivation/reperfusion (OGD/R); further experimental evidence showed that the protective role was limited in the PI3K inhibitor (LY294002) treatment group. Our results demonstrated that the functional monomer of B. rapa L. exerted a neuroprotective effect against OGD/R-induced HT22 cell injury, and its potential mechanism provides a scientific basis for future clinical applications and its use as a functional food.
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Brassica rapa , Fármacos Neuroprotetores/uso terapêutico , Extratos Vegetais/uso terapêutico , Animais , Isquemia Encefálica/tratamento farmacológico , Linhagem Celular/efeitos dos fármacos , Hipocampo/citologia , Humanos , Fármacos Neuroprotetores/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Fitoterapia , Extratos Vegetais/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Traumatismo por Reperfusão/metabolismo , TibetRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: The Tibetan turnip (Brassica rapa L.) has a wide array of medicine properties including heat-clearing, detoxifying and anti-hypoxia as listed in the famous centuries-old Tibetan medicine classic "The Four Medical Tantras". Evidence-based medicine also indicated the anti-hypoxic effect of turnips, suggesting a potential link to neuroprotective effect on ischemic stroke. This thereby enables turnips to serve as a novel nontoxic agent in related treatment. AIM OF THE STUDY: This study aimed to investigate the neuroprotective effect and elucidate the mechanism of aqueous extract of turnip (AET) on cerebral ischemia/reperfusion. MATERIALS AND METHODS: The experimental models of cerebral ischemia included transient middle cerebral artery occlusion/reperfusion (MCAO) in C57BL/6J mice and oxygen-glucose deprivation/reoxygenation (OGD/R) in HT-22â¯cells. Long-term effect of AET on infarct volume was evaluated by microtubule-associated protein 2 (MAP2) immunofluorescence 28 days after MCAO, and on neurofunctional outcomes determined by rotarod, grid walking, and cylinder tests in the meantime. Efficacy of AET was determined by the cell viability, the release of lactate dehydrogenase (LDH) and reactive oxygen species (ROS) in neurons. The underlying mechanism of AET rescued OGD/R cells were characterized by PI3K, Akt and mTOR expressions, which were further used to validate AET's role in the pathway. RESULTS: AET can reduce cerebral infarct volume and ameliorate behavioral deficits of MCAO/R mice dose-dependently. In vitro experiment further demonstrated that suitable concentrations of AET inhibited ROS, LDH production and restored mitochondrial expression induced by OGD/R. AET pretreatment can reverse the OGD/R-induced decreased level of phosphorylation of PI3K, Akt, mTOR, whereas this effect was blocked in the LY294002 (PI3K inhibitor) treatment group. CONCLUSIONS: AET improved the survival of OGD/R-injured HT-22â¯cells by activating the PI3K/Akt/mTOR pathway. Based on the results above, aqueous extract of turnip has a protective effect on focal cerebral ischemic injury.
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Isquemia Encefálica/tratamento farmacológico , Brassica rapa/química , Fármacos Neuroprotetores/farmacologia , Extratos Vegetais/farmacologia , Animais , Isquemia Encefálica/patologia , Linhagem Celular , Modelos Animais de Doenças , Infarto da Artéria Cerebral Média , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fármacos Neuroprotetores/isolamento & purificação , Proteína Oncogênica v-akt/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Traumatismo por Reperfusão/tratamento farmacológico , Traumatismo por Reperfusão/patologia , Serina-Treonina Quinases TOR/metabolismo , TibetRESUMO
For the immense requirement on agriculture and animal husbandry, application of pesticides and veterinary drugs had become a normal state in the farming and ranching areas. However, to intently pursue the yields, large quantities of residues of pesticides and veterinary drugs have caused serious harm to both the environment and the food industry. To control and solve such an issue, a variety of novel techniques were developed in recent years. In this review, the development and features about point-of-care-testing (POCT) detection on the residues of pesticides and veterinary drugs, such as, electrochemistry (EC), enzyme-linked immunosorbent assay (ELISA) and nano-techniques, were systematically introduced. For each topic, we first interpreted the strategies and detailed account of such technical contributions on detection and assessment of the residues. Finally, the advantages and perspectives about mentioned techniques for ultrasensitive assessment and sensing on pesticides and veterinary drugs were summarized.
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Resíduos de Praguicidas , Praguicidas , Drogas Veterinárias , Animais , Contaminação de Alimentos/análise , Humanos , Resíduos de Praguicidas/análise , Praguicidas/análise , Drogas Veterinárias/análiseRESUMO
Torularhodin, extracted from Sporidiobolus pararoseus, is a significant carotenoid that is similar to lycopene in structure. Some studies have indicated torularhodin as having antioxidative activities. However, it has not been thoroughly studied with respect to its antioxidative activity and molecular mechanisms in liver injury. Therefore, the aim of this study was to elucidate the antioxidative activity of torularhodin against hydrogen peroxide- (H2O2-) induced damage and the mechanism involved through transcriptome analysis and to explore its antioxidant potential. BRL cells were first subjected to H2O2 damage and then treated with torularhodin. The results showed that at 10-5 g/ml, torularhodin had significant protective effects against H2O2-induced oxidative damage. Morphological and immunofluorescence staining showed that torularhodin could maintain cell integrity and enhance the activity of antioxidant enzymes in the cells. According to transcriptome analysis, 2808 genes were significantly differentially expressed (1334 upregulated and 1474 downregulated) after torularhodin treatment. These genes were involved in three major Gene Ontology categories (biological process, cellular component, and molecular function). Moreover, torularhodin was involved in some cellular pathways, such as cancer inhibition, antioxidation, and aging delay. Our data highlighted the importance of multiple pathways in the antioxidative damage of liver treated with torularhodin and will contribute to get the molecular mechanisms of torularhodin inhibition of hepatic oxidative damage.
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Carotenoides/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Basidiomycota/química , Basidiomycota/metabolismo , Carotenoides/química , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Perfilação da Expressão Gênica , Hepatócitos/citologia , Hepatócitos/metabolismo , Peróxido de Hidrogênio/farmacologia , RatosRESUMO
Alcoholic liver disease (ALD) is a major cause of liver injury worldwide. Oxidative damage is one of the main injuries caused by ALD. The aim of this study was to elucidate the preventive effects of torularhodin, extracted from Sporidiobolus pararoseus, on alcoholic liver injury in mice. The mechanisms involved were investigated using transcriptome analysis. Torularhodin supplementation decreased ethanol-induced aspartate transaminase (ALT), aspartate transaminase (AST) and low density lipoprotein (LDL) levels, and increased high density lipoprotein (HDL) levels in the serum of mice. In liver tissue, treatment with torularhodin increased ethanol-induced superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels and decreased tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) levels. Histological analysis showed that torularhodin could alleviate the negative effects of alcohol on the liver. Transcriptomic analysis showed that 806 genes were significantly differentially expressed (506 up-regulated and 300 down-regulated) after torularhodin treatment. These genes were involved in three main Gene Ontology categories (biological process, cellular component, and molecular function) and multiple pathways. Therefore, torularhodin was considered to have potential as a protective agent against ALD.
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Carotenoides/farmacologia , Hepatopatias Alcoólicas/tratamento farmacológico , Fígado/efeitos dos fármacos , Transcriptoma , Animais , Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Aspartato Aminotransferases/metabolismo , Basidiomycota/metabolismo , Etanol/efeitos adversos , Glutationa Peroxidase/metabolismo , Inflamação , Interleucina-1beta/metabolismo , Lipoproteínas LDL/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Estresse Oxidativo/efeitos dos fármacos , Farmacologia , Substâncias Protetoras/farmacologia , Superóxido Dismutase/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Ferric ions (Fe3+ ions) and l-cysteine (( l-Cys) in the human body have always played an irreplaceable role in biological processes, and overload or deficiency of Fe3+ ions and l-Cys in the biological system leads to various diseases. In this work, N,S-co-doped red-emitting carbon dots (R-CDs) were synthesized by a facile hydrothermal method. Because the doping of N and S gives a unique functional group distribution on the surface of R-CDs, it can be complexed with Fe3+ ions to construct an energy transfer quenching system. However, the presence of l-Cys competitively binds to Fe3+ ions, thus resulting in the photoluminescence recovery of R-CDs. Therefore, a "switch-on" dual function sensing platform has successfully been developed based on R-CDs for rapid identification and quantification of Fe3+ ions and l-Cys. The linear detection range of Fe3+ ions is 0-30 µM (limit of detection (LOD): 0.27 µM) and that of l-Cys is 0-24 µM (LOD: 0.14 µM). The sensor platform was used to detect Fe3+ ions and l-Cys in human serum samples with satisfactory results. Compared with traditional detection methods, this method is more time-saving and efficient and can be completed in 3 min. It is worth mentioning that the R-CDs not only has high optical stability but also has negligible cytotoxicity and has been successfully applied to in vitro/vivo imaging, indicating that R-CDs have excellent tissue penetration and biomarker potential. More interestingly, the switch-on fluorescence behavior for stepwise detection of Fe3+ ions and l-Cys can also be observed in cell imaging, which provides the possibility of visual detection of the probe to be applied in vivo.
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Torularhodin is a natural product extracted from Sporidiobolus pararoseus and has a similar chemical structure to ß-carotene. The antioxidative effects of torularhodin were investigated using DPPH, ABTS, a cell oxidative damage model in vitro, and a d-galactose-induced liver-injured mouse model in vivo. Cell experiments demonstrated that torularhodin had a powerful effect on oxidative damage caused by H2O2 to AML12 cells. Torularhodin significantly reduced inflammatory cytokines and increased the activity of antioxidant enzymes both in mouse serum and the liver. The inhibition of d-galactose-induced oxidative damage in the liver was correlated with the torularhodin-mediated effects on improving the activity of Nrf2/HO-1, reducing the expression of Bax and NF-κB p65 by western blot analysis. RT-PCR results demonstrated torularhodin upregulated the antioxidative mRNA expression of Nrf2, NQO1, and HO-1 in the liver. In summary, torularhodin significantly scavenged free radicals and prevented oxidative damage in vitro and reduced d-galactose-induced liver oxidation via promotion of the Nrf2/HO-1 pathways in vivo.
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Antioxidantes/administração & dosagem , Basidiomycota/química , Carotenoides/administração & dosagem , Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Animais , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Galactose/efeitos adversos , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Humanos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos ICR , NAD(P)H Desidrogenase (Quinona)/genética , NAD(P)H Desidrogenase (Quinona)/metabolismo , Fator 2 Relacionado a NF-E2/genética , NF-kappa B/genética , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacosRESUMO
An ON-OFF-ON dual-function fluorescent nanoprobe is described for the trace detection of ferric ions and inositol hexaphosphate (IP6) in living cells. It is based on the use of yellow-fluorescent nitrogen-doped carbon dots (YN-CDs). Highly fluorescent YN-CDs were synthesized by a hydrothermal process. They have an absolute quantum yield of 2.15% and excitation/emission peaks at 420/575 nm. Fluorescence is quenched by Fe3+via photo-induced electron transfer. The quenchometric assay has a 34 nM detection limit for Fe(iii). On addition of IP6 which has a high affinity for Fe3+ due to the formation of Fe-O-P bonds, fluorescence becomes gradually restored. The resulting ON-OFF-ON assays for Fe(iii) and IP6 are reliable and sensitive. IP6 can be detected at concentrations as low as 2 nM. The nanoprobe was then applied to the determination of Fe3+ and IP6 in living cells in a food matrix. Furthermore, YN-CDs exhibited excellent biocompatibility. Hence, the probe can be applied as a fluorescent ink for bioimaging, both in vitro (cancer cells and bacteria) and in vivo (nematodes and mice).
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Ferro/análise , Ácido Fítico/análise , Pontos Quânticos/química , Animais , Apium/química , Carbono/química , Grão Comestível/química , Escherichia coli , Fluorescência , Análise de Alimentos , Frutas/química , Células Hep G2 , Humanos , Limite de Detecção , Camundongos Nus , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Nematoides , Espectrometria de Fluorescência/métodosRESUMO
: Aloe vera is a medicinal plant species of the genus Aloe with a long history of usage around the world. Acemannan, considered one of the main bioactive polysaccharides of Aloe vera, possesses immunoregulation, anti-cancer, anti-oxidation, wound healing and bone proliferation promotion, neuroprotection, and intestinal health promotion activities, among others. In this review, recent advancements in the extraction, purification, structural characteristics and biological activities of acemannan from Aloe vera were summarized. Among these advancements, the structural characteristics of purified polysaccharides were reviewed in detail. Meanwhile, the biological activities of acemannan from Aloe vera determined by in vivo, in vitro and clinical experiments are summarized, and possible mechanisms of these bioactivities were discussed. Moreover, the latest research progress on the use of acemannan in dentistry and wound healing was also summarized in details. The structure-activity relationships of acemannan and its medical applications were discussed. Finally, new perspectives for future research work on acemannan were proposed. In conclusion, this review summarizes the extraction, purification, structural characteristics, biological activities and pharmacological applications of acemannan, and provides information for the industrial production and possible applications in dentistry and wound healing in the future.
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Aloe/química , Mananas/química , Mananas/isolamento & purificação , Animais , Humanos , Mananas/farmacologia , Plantas Medicinais/química , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Relação Estrutura-Atividade , Cicatrização/efeitos dos fármacosRESUMO
Lipopolysaccharide (LPS) is the main virulence factor of Gram-negative bacteria, which can incite inflammation in tissues by inducing cells to secrete a variety of proinflammatory mediators, including cytokines, chemokines, interleukins, and prostaglandins. Herein, we chose LPS as an inducer to establish an inflammatory model of HeLa cells, and explored the effects of LPS on energy metabolism. We treated HeLa cells with different concentrations (0, 0.4, 1.0, 2.0, 4.0, and 6.0 µg/mL) of LPS for 24 h, and explored its effects on intercellular adenosine triphosphate (ATP) levels, intercellular nitrous oxide (NO) content, mitochondrial functions, and enzyme activities related to energy metabolism. Furthermore, we used metabonomics to study the metabolites that participated in energy metabolism. We found a positive correlation between LPS concentrations and intracellular ATP levels. In addition, LPS increased intracellular NO production, altered mitochondrial functions, strengthened glycolytic enzyme activities, and changed metabolites related to energy metabolism. Hence, in this study, we showed that LPS can strengthen energy metabolism by enhancing glycolysis, which could be used as an early diagnostic biomarker or a novel therapeutic target for inflammation-associated cancers.
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Metabolismo Energético/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Trifosfato de Adenosina/metabolismo , Células HeLa , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/fisiologia , Neoplasias/metabolismo , Óxido Nítrico/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
Antioxidant evaluation of bioactive compounds is limited, since many methods lack a real physiological environment that can be used conveniently and intuitively. In this study, a simple, label-free and effective electrochemical biosensor method has been developed to evaluate the antioxidant effect of phloretin (Ph) by 3D cell modification on a glassy carbon electrode (GCE). In response to this, A549 cells were immobilized onto a self-assembled Ê-cysteine/gold nanoparticle (AuNPs/Ê-Cys)-modified GCE surface by a simple drop casting after encapsulated in alginate. The electrochemical impedance spectroscopy (EIS) results showed that the impedance value (Ret) increased with the concentration of H2O2 in the range of 0-60⯵mol/L with the correlation of 0.990 which acted as an oxidative stress model inducer. However, the EIS value decreased with the co-incubation of Ph ranging from 10 to 100⯵mol/L, showing a dose-dependent manner and time effect, indicating that the variation of Ret was responded to the antioxidant effect. The response impedance of the biosensor is linear to Ph concentrations from 20⯵mol/L to 100⯵mol/L with the detection limit (LOD) as 1.96⯵mol/L. A significant correlation was observed between reactive oxygen species (ROS) values and Ret values following the concentrations of Ph, thus demonstrating the good biological relevance of cell-based electrochemical method. The strategy has been used to evaluate Ph antioxidant capacity in real cells with satisfactory results, indicating the feasibility of biosensor analysis for antioxidant evaluation.
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Técnicas Biossensoriais/métodos , Carbono/química , Técnicas Eletroquímicas , Eletrodos , Nanopartículas Metálicas/química , Floretina/análise , Células A549 , Ácido Algínico/química , Técnicas Biossensoriais/instrumentação , Cisteína/química , Ouro/química , Humanos , Peróxido de Hidrogênio/química , Limite de DetecçãoRESUMO
MicroRNAs (miRNAs), a kind of single-stranded small RNA molecule, play significant roles in the physiological and pathological processes of human beings. Currently, miRNAs have been demonstrated as important biomarkers critically related to many diseases and life nature, including several cancers and cell senescence. It is valuable to establish sensitive assays for monitoring the levels of intracellular up-regulated/down-regulated miRNA expression, which would contribute to the early prediction of the tumor risk and cardiovascular disease. Here, an oriented gold nanocross (AuNC)-decorated gold nanorod (AuNR) probe with "OFF-enhanced ON" fluorescence switching was developed based on fluorescence resonance energy transfer and surface enhanced fluorescence (FRET-SEF) principle. The nanoprobe was used to specifically detect miRNA in vitro, which gave two linear responses represented by the equation F = 1830.32 log C + 6349.27, R2 = 0.9901, and F = 244.41 log C + 1916.10, R2 = 0.9984, respectively, along with a detection limit of 0.5 aM and 0.03 fM, respectively. Furthermore, our nanoprobe was used to dynamically monitor the expression of intracellular up-regulated miRNA-34a from the HepG2 and H9C2 cells stimulated by AFB1 and TGF-ß1, and the experimental results showed that the new probe not only could be used to quantitively evaluate miRNA oncogene in vitro, but also enabled tracking and imaging of miRNAs in living cells.