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Purpose: Pancreatic adenocarcinoma (PAAD) presents an extremely high morbidity and mortality rate. Broccoli has excellent anti-cancer properties. However, the dosage and serious side effects still limit the application of broccoli and its derivatives for cancer therapy. Recently, extracellular vesicles (EVs) derived from plants are emerging as novel therapeutic agents. Thus, we conducted this study to determine the effectiveness of EVs isolated from Se-riched broccoli (Se-BDEVs) and conventional broccoli (cBDEVs) for the treatment of PAAD. Methods: In this study, we first isolated Se-BDEVs and cBDEVs by a differential centrifugation method, and characterized them by using nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Then, miRNA-seq was combined with target genes prediction, and functional enrichment analysis to reveal the potential function of Se-BDEVs and cBDEVs. Finally, the functional verification was conducted in PANC-1 cells. Results: Se-BDEVs and cBDEVs exhibited similar characteristics in size and morphology. Subsequent miRNA-seq revealed the expression of miRNAs in Se-BDEVs and cBDEVs. Using a combination of miRNA target prediction and KEGG functional analysis, we found miRNAs in Se-BDEVs and cBDEVs may play an important role in treating pancreatic cancer. Indeed, our in vitro study showed that Se-BDEVs had greater anti-PAAD potency than cBDEVs due to increased bna-miR167a_R-2 (miR167a) expression. Transfection with miR167a mimics significantly induced apoptosis of PANC-1 cells. Mechanistically, further bioinformatics analysis showed that IRS1, which is involved in the PI3K-AKT pathway, is the key target gene of miR167a. Conclusion: This study highlights the role of miR167a transported by Se-BDEVs which could be a new tool for counteracting tumorigenesis.
Assuntos
Adenocarcinoma , Brassica , Vesículas Extracelulares , MicroRNAs , Neoplasias Pancreáticas , Selênio , Humanos , Brassica/genética , Brassica/metabolismo , Selênio/uso terapêutico , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/tratamento farmacológico , Adenocarcinoma/tratamento farmacológico , Biofortificação , Fosfatidilinositol 3-Quinases/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Vesículas Extracelulares/metabolismo , Apoptose , Proteínas Substratos do Receptor de Insulina/metabolismo , Neoplasias PancreáticasRESUMO
Imbalanced nutrient intake causes abnormal energy metabolism, which results in obesity. There is feasible evidence that selenium-rich (Se-rich) foods may alleviate obesity and enhance general public health, but the underlying mechanisms remain elusive. Herein we examined the effect of Se supplementation on white adipose tissue beiging process. The mice were fed with a normal diet or a Se-deficient high-fat diet (DHFD) until significant differences in terms of body weight, glucose tolerance and insulin sensitivity. Next, mice in the DHFD group were changed to a high-fat diet (HFD) containing specified amounts of selenomethionine (SeMet) (0, 150, 300, and 600 µg/kg) and continued to feed for 14 weeks. Notably, 150 µg/kg SeMet supplement highly protected mice from DHFD-induced obesity, insulin resistance, and lipid deposits in the liver and kidney, and featured by the enhanced beiging process in white adipose tissue and increased energy expenditure. Moreover, upon cold challenge, 150 µg/kg SeMet supplement enhanced cold tolerance in mice by inducing adipose beiging to promote energy expenditure, as evidenced by the increased expression of uncoupling protein-1 (UCP1) in adipocytes. Similarly, SeMet (10 µM) promoted the differentiation of beige adipocytes from the stromal vascular fraction. Collectively, our data support that optimal supplementation of SeMet could enhance the beiging process to attenuate HFD-induced obesity, which provides new insights into the relationship between dietary SeMet and type 2 diabetes.
Assuntos
Diabetes Mellitus Tipo 2 , Resistência à Insulina , Camundongos , Animais , Selenometionina/farmacologia , Diabetes Mellitus Tipo 2/metabolismo , Tecido Adiposo Branco/metabolismo , Obesidade/etiologia , Obesidade/prevenção & controle , Obesidade/metabolismo , Dieta Hiperlipídica/efeitos adversos , Metabolismo Energético , Camundongos Endogâmicos C57BL , Tecido Adiposo/metabolismoRESUMO
Aberrant amino acid metabolism is a common event in obesity. Particularly, subjects with obesity are characterized by the excessive plasma kynurenine (Kyn). However, the primary source of Kyn and its impact on metabolic syndrome are yet to be fully addressed. Herein, we show that the overexpressed indoleamine 2,3-dioxygenase 1 (IDO1) in adipocytes predominantly contributes to the excessive Kyn, indicating a central role of adipocytes in Kyn metabolism. Depletion of Ido1 in adipocytes abrogates Kyn accumulation, protecting mice against obesity. Mechanistically, Kyn impairs lipid homeostasis in adipocytes via activating the aryl hydrocarbon receptor (AhR)/Signal transducer and activator of transcription 3 /interleukin-6 signaling. Genetic ablation of AhR in adipocytes abolishes the effect of Kyn. Moreover, supplementation of vitamin B6 ameliorated Kyn accumulation, protecting mice from obesity. Collectively, our data support that adipocytes are the primary source of increased circulating Kyn, while elimination of accumulated Kyn could be a viable strategy against obesity.
Assuntos
Resistência à Insulina , Cinurenina , Adipócitos/metabolismo , Animais , Humanos , Indolamina-Pirrol 2,3,-Dioxigenase/metabolismo , Interleucina-6/metabolismo , Cinurenina/metabolismo , Camundongos , Obesidade , Receptores de Hidrocarboneto Arílico/genética , Receptores de Hidrocarboneto Arílico/metabolismo , Fator de Transcrição STAT3/metabolismo , Triptofano Oxigenase/metabolismoRESUMO
The prevalence and mortality rate of colorectal cancer (CRC) have been increasing dramatically worldwide. Pinus massoniana pollen, a well-known natural food, is one of the most commonly consumed traditional medicines in China. P. massoniana pollen polysaccharides (PPPS) have antitumor effects, but it remains unclear whether they can inhibit CRC. Here, we have demonstrated that PPPS inhibited CRC cell proliferation effectively, induced morphology changes, triggered apoptosis by upregulating key apoptosis-related proteins, and arrested the cell cycle at the G0/G1 phase. Moreover, PPPS markedly inhibited CRC cell metastasis by downregulating MMP-9 and inhibiting epithelial-mesenchymal transition. In vivo, PPPS exhibited potent antitumor activity and no observable toxicity in BALB/c nude mice bearing HCT-116 tumors. Most strikingly, PPPS pre-treatment dramatically inhibited the growth of incipient tumors, although not as effectively as in the PPPS-Ther group. Thus, our results suggest that PPPS can be a potential anti-CRC agent, paving the way for developing complex carbohydrates for tumor prevention and treatment.
Assuntos
Neoplasias Colorretais , Pinus , Animais , Apoptose , Proliferação de Células , Neoplasias Colorretais/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Pólen , Polissacarídeos/farmacologiaRESUMO
Oxidative stress is responsible for the pathogeneses of various diseases. Mitochondrial dysfunction, impaired DNA repair, and cellular damage followed by oxidative stress contribute to neurodegenerative diseases, such as Alzheimer disease (AD) and Parkinson disease (PD). Acupuncture is a traditional therapy that has been practiced for >3000 years in Asia. Many studies have demonstrated that acupuncture has notable antioxidative, anti-inflammatory, and antiapoptotic effects. However, the exact mechanism remains unclear. Nuclear factor erythroid 2-related factor (Nrf2) is crucial in regulating the redox equilibrium. Activated Nfr2 translocates into the nucleus, binds to the antioxidant response element (ARE), and initiates antioxidative enzyme transcription. In this review, we demonstrated the effects of acupuncture on oxidative stress amelioration in AD and PD animal models through Nrf2/ARE pathway activation and Nrf2/ARE-related pathway regulation. Thus, acupuncture could be a therapeutic option for AD and PD.
RESUMO
Herein, we review the characteristics of the six predominant venomous snakes in Taiwan and the effects of traditional Chinese medicine on the long-term outcomes of snakebite venom. We electronically searched databases, including PubMed, ClinicalKey, China National Knowledge Infrastructure, National Digital Library of Theses and Dissertations in Taiwan, and Airiti Library, from their inception to November 2019 by using the following Medical Subject Headings' keywords: snakebite, long-term, chronic, Chinese medicine, CAM, herb, and Taiwan. The most common long-term effects of snakebite envenomation include "migraine-like syndrome", brain injuries caused by hypoxia or intracranial hemorrhage, and chronic kidney disease. In addition, hypopituitarism is also worth mentioning. Traditional Chinese medicine can potentially be used in a complementary or alternative treatment for these effects, but additional studies are needed.
Assuntos
Encefalopatias/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Hipopituitarismo/tratamento farmacológico , Medicina Tradicional Chinesa , Insuficiência Renal Crônica/tratamento farmacológico , Mordeduras de Serpentes/complicações , Animais , Encefalopatias/etiologia , Infarto Cerebral/tratamento farmacológico , Infarto Cerebral/etiologia , Medicamentos de Ervas Chinesas/administração & dosagem , Humanos , Hipopituitarismo/etiologia , Hemorragias Intracranianas/tratamento farmacológico , Hemorragias Intracranianas/etiologia , Transtornos de Enxaqueca/tratamento farmacológico , Transtornos de Enxaqueca/etiologia , Insuficiência Renal Crônica/etiologia , TaiwanRESUMO
BACKGROUND: Injection of exogenous hyaluronic acid (HA) into the joint capsule improves symptoms of early stage osteoarthritis (OA). However, reactive oxygen species degrade HA into small oligosaccharides that can elicit pro-inflammatory responses. Likewise, disturbance of the antioxidant enzyme system and generation of oxidative stress by pro-inflammatory cytokines worsen knee OA. Accordingly, we proposed the use of aucubin, an antioxidant and anti-inflammatory compound, as a versatile adjuvant to HA for treating OA. METHODS: Primary human chondrocytes were cultured in media supplemented with aucubin in a series of concentrations (0, 0.01, 0.1, 1, and 10⯵g/ml) to study dose-dependent toxicity. We then evaluated the therapeutic effects of HA (100⯵g/ml) supplemented with aucubin (10⯵g/ml) on interleukin-1 beta (IL-1ß, 10â¯ng/ml)-stimulated chondrocytes. RESULTS: The use of aucubin did not change cell viability or alter lactate dehydrogenase release to normal chondrocytes. Although the proliferation and sulfated glycosaminoglycan production were not affected, aucubin partially restored the hypertrophic transformation of chondrocytes. Relative to treatment with HA or aucubin alone, real-time PCR revealed that aucubin-supplemented HA down-regulated the mRNA levels of tumor necrosis factor-alpha (TNF-α), corrected collagen type 1 and aggrecan, and up-regulated tissue inhibitor of metalloproteinase 1. Moreover, ELISA testing also showed a reduced TNF-α production. Although superoxide dismutases activity was still distributed, aucubin restored total antioxidant capacity of IL-1ß-stimulated chondrocytes. Western blotting further showed that aucubin inhibited cyclooxygenase-2 and regulated the nuclear factor (erythroid-derived 2)-like 2 pathway. CONCLUSION: Aucubin can enhance the anti-catabolic and anti-inflammatory effects of HA on OA chondrocytes.
Assuntos
Condrócitos/efeitos dos fármacos , Ácido Hialurônico/farmacologia , Glucosídeos Iridoides/farmacologia , Osteoartrite do Joelho/patologia , Antioxidantes/farmacologia , Cartilagem Articular/patologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Condrócitos/metabolismo , Condrócitos/patologia , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Ciclo-Oxigenase 2/metabolismo , Citocinas/genética , Citocinas/metabolismo , Glicosaminoglicanos/metabolismo , Humanos , Interleucina-1beta/farmacologia , Glucosídeos Iridoides/administração & dosagem , Fator de Necrose Tumoral alfa/genéticaRESUMO
Three new anthraquinones, lasianthurin B (1), C (2), lasianthuoside D (3), a new benzochromene, lasianthurin D (4), and a new furfural glycoside, lasianthuoside E (5), together with one known compound 4- hydroxymethyl-2-furaldehyde (6) were isolated from an alcohol extract of the root of Lasianthus acuminatissimus. Their structures were elucidated on the basis of extensive spectroscopic data analysis (including 1D, 2D NMR, X-ray, and MS experiments) and comparsion to literature data.
Assuntos
Antraquinonas/química , Benzopiranos/química , Furaldeído/química , Glicosídeos/química , Rubiaceae/química , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Extratos Vegetais/química , Raízes de Plantas/química , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
This study investigated the polyphenol content, antioxidant activity, and inhibition ability of mushroom tyrosinase and melanogenesis of Dendrobium tosaense (DT) extract. Ground DT was extracted using deionized water (W) or 50% ethanol (50E) at room temperature (RT) or 50 °C (50T) for 20 min. The 50T + 50E extract exhibited the highest total phenol content 47.0 ± 4.0 mg gallic acid equivalent/g DT extract, the highest level of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) free-radical scavenging 66.0 ± 3.0 mg Trolox equivalent/g DT extract, and the highest reducing power 12.00 ± 0.50 mg vitamin C equivalent/g DT extract. The RT + W extract had the highest total flavonoid content 110.0 ± 3.0 mg quercetin equivalent/g DT extract. The RT + 50E extract had the lowest half maximal inhibitory concentration 1.30 ± 0.00 mg/mL for 2,2-diphenyl-1-picrylhydrazyl free-radical scavenging, and the lowest half maximal inhibitory concentration 6.40 ± 0.30 mg/mL for mushroom tyrosinase inhibition activity. DT extracts, especially RT + W and 50T + W, exhibited potent inhibitory effects on melanogenesis of B16/F10 cells. These results demonstrated the application potential of DT extract for skincare.
Assuntos
Antioxidantes/química , Antioxidantes/farmacologia , Dendrobium/química , Melaninas/biossíntese , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Agaricales/enzimologia , Animais , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida , Sequestradores de Radicais Livres/química , Sequestradores de Radicais Livres/farmacologia , Melanoma Experimental , Camundongos , Monofenol Mono-Oxigenase/antagonistas & inibidores , Oxirredução/efeitos dos fármacos , Compostos Fitoquímicos/química , Espectrometria de Massas em TandemRESUMO
OBJECTIVES: To compare diffusion tensor (DT)-derived indices from the thalamic nuclei and cerebrospinal fluid (CSF) hydrodynamic parameters for the prediction of gait responsiveness to the CSF tap test in early iNPH patients. METHODS: In this study, 22 patients with iNPH and 16 normal controls were enrolled with the approval of an institutional review board. DT imaging and phase-contrast magnetic resonance imaging were performed in patients and controls to determine DT-related indices of the sensorimotor-related thalamic nuclei and CSF hydrodynamics. Gait performance was assessed in patients using gait scale before and after the tap test. The Mann-Whitney U test and receiver operating characteristic (ROC) curve analysis were applied to compare group differences between patients and controls and assess the predictive performance of gait responsiveness to the tap test in the patients. RESULTS: Fractional anisotropy (FA) and axial diffusivity showed significant increases in the ventrolateral (VL) and ventroposterolateral (VPL) nuclei of the iNPH group compared with those of the control group (p < 0.05). The predictions of gait responsiveness of ventral thalamic FA alone (area under the ROC curve [AUC] < 0.8) significantly outperformed those of CSF hydrodynamics alone (AUC < 0.6). The AUC curve was elevated to 0.812 when the CSF peak systolic velocity and FA value were combined for the VPL nucleus, yielding the highest sensitivity (0.769) and specificity (0.778) to predict gait responses. CONCLUSIONS: Combined measurements of sensorimotor-related thalamic FA and CSF hydrodynamics can provide potential biomarkers for gait response to the CSF tap test in patients with iNPH. KEY POINTS: ⢠Ventrolateral and ventroposterolateral thalamic FA may predict gait responsiveness to tap test. ⢠Thalamic neuroplasticity can be assessed through DTI in idiopathic normal-pressure hydrocephalus. ⢠Changes in the CST associated with gait control could trigger thalamic neuroplasticity. ⢠Activities of sensorimotor-related circuits could alter in patients with gait disturbance. ⢠Management of patients with iNPH could be more appropriate.
Assuntos
Líquido Cefalorraquidiano/fisiologia , Marcha/fisiologia , Hidrocefalia de Pressão Normal/fisiopatologia , Tálamo/fisiologia , Idoso , Anisotropia , Estudos de Casos e Controles , Imagem de Difusão por Ressonância Magnética , Imagem de Tensor de Difusão , Feminino , Humanos , Hidrodinâmica , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Curva ROC , Sensibilidade e EspecificidadeRESUMO
Proinflammatory cytokines and reactive oxygen species (ROS) are known to be involved in the initiation and progression of osteoarthritis (OA). New evidence clarifying the correlation between ROS and inflammation has indicated that oxidative stress can up-regulate inflammatory cytokines. l-Ascorbic acid (AA), an antioxidant, has been shown to have anti-inflammatory effects and improve matrix deposition in chondrocytes. The purpose of this study was to examine the effects of hyaluronic acid (HA; 100 µg/mL) supplemented with AA (50 µg/mL) on human normal and interleukin-1 beta-stimulated (IL-1ß, 10 ng/mL) chondrocytes. HA, AA, and HA + AA treatment did not change cell morphology, viability, proliferation, and glycosaminoglycan production in normal chondrocytes. HA, AA, and HA + AA, by contrast, partially restored viability and morphology of hypertrophic chondrocytes, and HA and HA + AA further decreased the cytotoxicity of IL-1ß. Real-time PCR revealed that AA and HA + AA had no substantial effects on unstimulated chondrocytes, except for down-regulation of matrix metalloproteinase (MMP)-9 mRNA levels. For IL-1ß-stimulated chondrocytes, significant down-regulation of IL-1ß, tumor necrosis factor-alpha (TNF-α), MMP-3, and MMP-9 mRNA expression was found when cells were cultured in HA-supplemented media. Moreover, HA + AA supplementation further significantly decreased MMP-3 and MMP-9 mRNA expression. The protein production of MMP-3 was decreased, with a significant difference between the HA + AA group and HA group. The antioxidant capacity and superoxide dismutases activity were also partially restored in stimulated chondrocytes. HA supplemented with AA modulates MMPs expression and antioxidant fuction in chondrocytes. AA may enhance the anticatabolic effects of HA on OA chondrocytes. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1809-1817, 2018.
Assuntos
Ácido Ascórbico/farmacologia , Condrócitos/enzimologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Ácido Hialurônico/farmacologia , Metaloproteinase 3 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Osteoartrite/metabolismo , Idoso , Idoso de 80 Anos ou mais , Antioxidantes/farmacologia , Ácido Ascórbico/agonistas , Condrócitos/patologia , Sinergismo Farmacológico , Feminino , Humanos , Ácido Hialurônico/agonistas , Masculino , Pessoa de Meia-Idade , Osteoartrite/tratamento farmacológico , Osteoartrite/patologia , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Reference proteins and biomarkers are important for the quantitative evaluation of protein abundance. Chlamydomonasreinhardtii was grown under five stress conditions (dark, cold, heat, salt, and glucose supplementation), and the OD750 and total protein contents were evaluated on days 0, 1, 2, 4, and 6 of culture. Antibodies for 20 candidate proteins were generated, and the protein expression patterns were examined by western blotting. Reference protein(s) for each treatment were identified by calculating the Pearson's correlation coefficient (PCC) between target protein abundance and total protein content. Histone H3, beta tubulin 1 (TUB-1), ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (RBCL), and mitochondrial F1F0 ATP synthase subunit 6 (ATPs-6) were the top reference proteins, because they were expressed stably under multiple stress conditions. The average relative-fold change (ARF) value of each protein was calculated to identify biomarkers. Heat shock protein 90B (HSP90B), flagellar associated protein (FAP127) and ATP synthase CF0 A subunit (ATPs-A) were suitable biomarkers for multiple treatments, while receptor of activated protein kinase C1 (RCK1), biotin carboxylase (BCR1), mitochondrial phosphate carrier protein (MPC1), and rubisco large subunit N-methyltransferase (RMT1) were suitable biomarkers for the dark, cold, heat, and glucose treatments, respectively.
Assuntos
Chlamydomonas reinhardtii/metabolismo , Proteínas de Plantas/metabolismo , Proteoma , Proteômica , Estresse Fisiológico , Biomarcadores , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Proteômica/métodosRESUMO
OBJECTIVE: To determine bacteriostatic abilities of Artemisia argyi extracts, and to explore the effect of Artemisia argyi extracts on oral ulcer in rats.â© Methods: We extracted the mixture of Artemisia argyi volatile oils and water-extraction by leaching method and evaluated the anti-microbial effect of Artemisia argyi extracts on common oral floras in vitro. The rat cheeks were burnt by NaOH to establish the models of oral ulcer. The curative effects of crude drug of Artemisia argyi extracts at 2.0, 1.0, 0.5 g/mL on oral ulcer in rats were evaluated by measuring the oral ulcer healing time. Serum TNF-α level and expression of proliferating cell nuclear antigen (PCNA) were analyzed by ELASA and immunohistochemical staining.â© Results: Artemisia argyi extracts obviously inhibited the Staphylococcus aureus and Streptococcus. NaOH-made oral ulcer in rats were successfully established. The crude drug at 2.0 and 1.0 g/mL obviously reduced healing time, significantly inhibited the release of TNF-α, and improved the PCNA level in the ulcer tissues (All P<0.01). The extracts obviously reduced the local inflammatory reaction and promoted tissue repair of oral ulcer.â© Conclusion: Artemisia argyi extracts promote tissue repair of oral ulcer via inhibiting bacterial growth, reducing the release of TNF-α and improving the PCNA level.
Assuntos
Artemisia/química , Úlceras Orais/tratamento farmacológico , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Animais , Anti-Infecciosos/farmacologia , Anti-Infecciosos/uso terapêutico , Inflamação/tratamento farmacológico , Óleos de Plantas/farmacologia , Óleos de Plantas/uso terapêutico , Ratos , Resultado do Tratamento , Cicatrização/efeitos dos fármacosRESUMO
Vacuoles play a fundamental role in storage and remobilization of various nutrients, including phosphorus (P), an essential element for cell growth and development. Cells acquire P primarily in the form of inorganic orthophosphate (Pi). However, the form of P stored in vacuoles varies by organism and tissue. Algae and yeast store polyphosphates (polyPs), whereas plants store Pi and inositol phosphates (InsPs) in vegetative tissues and seeds, respectively. In this review, we summarize how vacuolar P molecules are stored and reallocated and how these processes are regulated and co-ordinated. The roles of SYG1/PHO81/XPR1 (SPX)-domain-containing membrane proteins in allocating vacuolar P are outlined. We also highlight the importance of vacuolar P in buffering the cytoplasmic Pi concentration to maintain cellular homeostasis when the external P supply fluctuates, and present additional roles for vacuolar polyP and InsP besides being a P reserve. Furthermore, we discuss the possibility of alternative pathways to recycle Pi from other P metabolites in vacuoles. Finally, future perspectives for researching this topic and its potential application in agriculture are proposed.
Assuntos
Proteínas de Membrana/genética , Fósforo/metabolismo , Proteínas de Plantas/genética , Plantas/metabolismo , Vacúolos/metabolismo , Leveduras/metabolismo , Proteínas de Membrana/metabolismo , Proteínas de Plantas/metabolismoRESUMO
The incidence of osteoporosis has increased among the elderly population. Establishing a model of bone remodeling for screening new drugs is critical to identify safe and effective treatments for osteoporosis. In this study, we established a platform to investigate the therapeutic effects of collagenous peptides extracted from scales of two kinds of fish, namely, sparidae and chanos. These peptides were prepared using seven concentrations of collagenous peptide: 100, 80, 60, 40, 20, 10 and 1 mg/ml. Experimental results indicated that collagenous peptides promoted the proliferation of osteoblasts and inhibited the proliferation of mature osteoclasts; the effective concentration of collagenous peptide-sparidae was 10 mg/ml and that of collagenous peptide-chanos was 40 mg/ml. These findings demonstrate that, to a certain extent, collagenous peptides extracted from fish scales can be used to prevent osteoporosis to assist bone remodeling.
Assuntos
Colágeno/uso terapêutico , Proteínas de Peixes/uso terapêutico , Osteoblastos/efeitos dos fármacos , Osteoclastos/efeitos dos fármacos , Osteoporose/prevenção & controle , Animais , Reabsorção Óssea/prevenção & controle , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Colágeno/farmacologia , Avaliação Pré-Clínica de Medicamentos , Proteínas de Peixes/farmacologia , Humanos , PerciformesRESUMO
Research has indicated that fatigue is a critical factor in cognitive lapses because it negatively affects an individual's internal state, which is then manifested physiologically. This study explores neurophysiological changes, measured by electroencephalogram (EEG), due to fatigue. This study further demonstrates the feasibility of an online closed-loop EEG-based fatigue detection and mitigation system that detects physiological change and can thereby prevent fatigue-related cognitive lapses. More importantly, this work compares the efficacy of fatigue detection and mitigation between the EEG-based and a nonEEG-based random method. Twelve healthy subjects participated in a sustained-attention driving experiment. Each participant's EEG signal was monitored continuously and a warning was delivered in real-time to participants once the EEG signature of fatigue was detected. Study results indicate suppression of the alpha- and theta-power of an occipital component and improved behavioral performance following a warning signal; these findings are in line with those in previous studies. However, study results also showed reduced warning efficacy (i.e. increased response times (RTs) to lane deviations) accompanied by increased alpha-power due to the fluctuation of warnings over time. Furthermore, a comparison of EEG-based and nonEEG-based random approaches clearly demonstrated the necessity of adaptive fatigue-mitigation systems, based on a subject's cognitive level, to deliver warnings. Analytical results clearly demonstrate and validate the efficacy of this online closed-loop EEG-based fatigue detection and mitigation mechanism to identify cognitive lapses that may lead to catastrophic incidents in countless operational environments.
Assuntos
Atenção/fisiologia , Encéfalo/fisiopatologia , Eletroencefalografia/métodos , Fadiga/diagnóstico , Fadiga/fisiopatologia , Retroalimentação Psicológica , Estimulação Acústica/métodos , Adulto , Ritmo alfa/fisiologia , Condução de Veículo/psicologia , Cognição/fisiologia , Fadiga/terapia , Estudos de Viabilidade , Retroalimentação Psicológica/fisiologia , Feminino , Humanos , Masculino , Tempo de Reação , Fatores de Tempo , Interface Usuário-Computador , Adulto JovemRESUMO
RIG-I (retinoic acid inducible gene-I) is a key mediator of antiviral immunity, able to couple detection of infection by RNA and DNA viruses to the induction of interferons. In the present study, a RIG-I gene from grass carp Ctenopharyngodon idella (CiRIG-I) was isolated and characterized. The full-length cDNA of CiRIG-I was of 3198 bp and encoded a polypeptide of 947 amino acids with an estimated molecular mass of 108,730 Da and a predicted isoelectric point of 5.85, including six main overlapping structural domains: two CARDs (caspase activation and recruitment domain), one ResIII (conserved restriction domain of bacterial type III restriction enzyme), one DEXDc (DEAD/DEAH box helicase domain), one HELICc (helicase superfamily c-terminal domain) and one RD (regulatory domain). The CiRIG-I mRNA was widespread expression in the tested 15 tissues by semi-quantitative RT-PCR (sqRT-PCR) assay. The CiRIG-I expressions in spleen and liver were significantly induced following grass carp reovirus (GCRV) infection. CiRIG-I mRNA expression was rapidly and significantly up-regulated in vitro after GCRV infection, and the CiRIG-I transcripts were also significantly enhanced in vitro post the synthetic double stranded RNA polyinosinic-polycytidylic potassium salt (poly(I:C)) stimulation. These results collectively suggested that CiRIG-I was an inducible protein, involved in the antiviral innate immune defense to GCRV in grass carp, and laid the foundation for the further mechanism research of RIG-I in fishes.
Assuntos
Carpas/genética , Carpas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Adjuvantes Imunológicos/farmacologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Carpas/classificação , Doenças dos Peixes/imunologia , Perfilação da Expressão Gênica , Imunidade Inata , Fígado/efeitos dos fármacos , Fígado/imunologia , Dados de Sequência Molecular , Filogenia , Poli I-C/farmacologia , Infecções por Reoviridae/imunologia , Infecções por Reoviridae/veterinária , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Baço/efeitos dos fármacos , Baço/imunologia , Fatores de Tempo , Fatores de Transcrição/químicaRESUMO
IPS-1 (interferon-ß promoter stimulator 1), also known as MAVS/VISA/Cardif, plays a central role in antiviral immunity. In this manuscript, we cloned and characterized IPS-1 from grass carp Ctenopharyngodon idella (designated as CiIPS-1). The CiIPS-1 cDNA is 2412 bp long and consists of a 5' untranslated region (UTR) of 124 bp, a 3' UTR of 497 bp with three cytokine RNA instability motifs (ATTTA) and a polyadenylation signal (AATAAA), and an open reading frame (ORF) of 1791 bp encoding a polypeptide of 596 amino acids with a calculated molecular mass of 64.1 kDa and a theoretical isoelectric point of 4.79. Structural analysis showed that the CiIPS-1 protein contained an N-terminal CARD (caspase activation and recruitment domain), a central proline-rich domain, a putative TRAF2-binding motif and a C-terminal transmembrane domain. Similarity analysis of the deduced amino acid sequence of the CiIPS-1 by MatGAT software revealed that the CiIPS-1 shared 27.8-76.4% identity and 47.4-85.2% similarity with other known piscine IPS-1 sequences. The CiIPS-1 mRNA was constitutively expressed in the examined tissues, higher in spleen, and was induced by grass carp reovirus (GCRV) injection by semi-quantitative RT-PCR assay. Quantitative real-time RT-PCR analysis revealed that the CiIPS-1 mRNA expression was rapidly and significantly up-regulated in vivo and in vitro after GCRV infection, and the CiIPS-1 transcripts were also significantly enhanced in vitro post the synthetic double stranded RNA polyinosinic-polycytidylic potassium salt (poly(I:C)) stimulation. These results indicated that CiIPS-1 was an inducible acute-phase protein and involved in the immune reaction to GCRV in grass carp.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Carpas , Clonagem Molecular , Regulação da Expressão Gênica/fisiologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA Complementar/genética , Dados de Sequência Molecular , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reoviridae , Infecções por Reoviridae/metabolismo , Infecções por Reoviridae/veterinária , Infecções por Reoviridae/virologiaRESUMO
LGP2 (laboratory of genetics and physiology 2), a homologue of RIG-I (Retinoic acid inducible gene-I) and MDA5 (Melanoma differentiation associated gene 5) without the CARD (caspase activation and recruitment domain) required for signaling, plays a pivotal role in modulating signaling by RIG-I and MDA5 for interferon (IFN) synthesis. In this study, a novel LGP2 gene from grass carp Ctenopharyngodon idella (designated as CiLGP2) was isolated and characterized. The full-length cDNA of CiLGP2 was of 2920 bp with five instability motifs (ATTTA). The open reading frame was of 2043 bp and encoded a polypeptide of 680 amino acids, including five main overlapping structural domains: two DEXDc (DEAD/DEAH box helicase domain), one ResIII (conserved restriction domain of bacterial type III restriction enzyme), one HELICc (helicase superfamily c-terminal domain) and one RD (regulatory domain). There was one more alpha-helix in the RD, compared with that in human. The CiLGP2 mRNA was ubiquitous expression in the tested tissues, was high level in spleen, skin, heart and intestine tissues, and was up-regulated by grass carp reovirus (GCRV) injection by semi-quantitative RT-PCR (sqRT-PCR) assay. The CiLGP2 expression in spleen was significantly up-regulated at 12 h (14.5 folds, P < 0.05), reached the crest at 24 h (19.0 folds, P < 0.05), and then dropped a little at 48 h (10.4 folds) post-injection of GCRV and kept this level in the following test period (P < 0.05). In liver, the temporal expression of CiLGP2 mRNA was significantly increased at 24 h (3.8 times, P < 0.05), reached peak at 48 h (10.7 times, P < 0.05), and then decreased a little bit at 72 h (5.8 times, P < 0.05) and kept this high level by the end of the test (P < 0.05). These results collectively suggested that CiLGP2 was a novel member of RLR gene family, engaging in the early stage of antiviral innate immune defense in grass carp, and laid the foundation for the further mechanism research of LGP2 in fishes.