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1.
BMC Cancer ; 21(1): 1278, 2021 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-34836510

RESUMO

BACKGROUND: Whereas there are many pharmacological interventions prescribed for patients with advanced anaplastic lymphoma kinase (ALK)- rearranged non-small cell lung cancer (NSCLC), comparative data between novel generation ALK-tyrosine kinase inhibitors (TKIs) remain scant. Here, we indirectly compared the efficacy and safety of first-line systemic therapeutic options used for the treatment of ALK-rearranged NSCLC. METHODS: We included all phase 2 and 3 randomised controlled trials (RCTs) comparing any two or three treatment options. Eligible studies reported at least one of the following outcomes: progression free survival (PFS), overall survival (OS), objective response rate (ORR), or adverse events of grade 3 or higher (Grade ≥ 3 AEs). Subgroup analysis was conducted according to central nervous system (CNS) metastases. RESULTS: A total of 9 RCTs consisting of 2484 patients with 8 treatment options were included in the systematic review. Our analysis showed that alectinib (300 mg and 600 mg), brigatinib, lorlatinib and ensartinib yielded the most favorable PFS. Whereas there was no significant OS or ORR difference among the ALK-TKIs. According to Bayesian ranking profiles, lorlatinib, alectinib 600 mg and alectinib 300 mg had the best PFS (63.7%), OS (35.9%) and ORR (37%), respectively. On the other hand, ceritinib showed the highest rate of severe adverse events (60%). CONCLUSION: Our analysis indicated that alectinib and lorlatinib might be associated with the best therapeutic efficacy in first-line treatment for major population of advanced NSCLC patients with ALK-rearrangement. However, since there is little comparative evidence on the treatment options, there is need for relative trials to fully determine the best treatment options as well as the rapidly evolving treatment landscape.

2.
Hepatology ; 2021 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-34435375

RESUMO

BACKGROUND & AIMS: Although the prevalence of nonalcoholic fatty liver disease (NAFLD) has risen dramatically to 25% of the adult population worldwide, there are as yet no approved pharmacological interventions for the disease due to uncertainty about the underlying molecular mechanisms. It is known that mitochondrial dysfunction is an important factor in the development of NAFLD. Mitochondrial antiviral signaling protein (MAVS) is a critical signaling adaptor for host defenses against viral infection. However, the role of MAVS in mitochondrial metabolism during NAFLD progression remains largely unknown. APPROACH & RESULTS: Based on expression analysis, we identified a marked downregulation of MAVS in hepatocytes during NAFLD progression. By employing MAVS global knockout and hepatocyte-specific MAVS knockout mice, we found that MAVS is protective against diet-induced NAFLD. MAVS deficiency induces extensive mitochondrial dysfunction during NAFLD pathogenesis which was confirmed as impaired mitochondrial respiratory capacity and membrane potential. Metabolomics data also showed the extensive metabolic disorders after MAVS deletion. Mechanistically, MAVS interacts with the N-terminal stretch of voltage-dependent anion channel 2 (VDAC2), which is required for the ability of MAVS to influence mitochondrial function and hepatic steatosis. CONCLUSIONS: In hepatocytes, MAVS plays an important role in protecting against NAFLD by helping to regulate healthy mitochondrial function. These findings provide new insights regarding the metabolic importance of conventional immune regulators and support the possibility that targeting MAVS may represent a new avenue for treating NAFLD.

3.
Cell Metab ; 33(8): 1640-1654.e8, 2021 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-34107313

RESUMO

Obesity is characterized by the excessive accumulation of the white adipose tissue (WAT), but healthy expansion of WAT via adipocyte hyperplasia can offset the negative metabolic effects of obesity. Thus, identification of novel adipogenesis regulators that promote hyperplasia may lead to effective therapies for obesity-induced metabolic disorders. Using transcriptomic approaches, we identified transmembrane BAX inhibitor motif-containing 1 (TMBIM1) as an inhibitor of adipogenesis. Gain or loss of function of TMBIM1 in preadipocytes inhibited or promoted adipogenesis, respectively. In vivo, in response to caloric excess, adipocyte precursor (AP)-specific Tmbim1 knockout (KO) mice displayed WAT hyperplasia and improved systemic metabolic health, while overexpression of Tmbim1 in transgenic mice showed the opposite effects. Moreover, mature adipocyte-specific Tmbim1 KO did not affect WAT cellularity or nutrient homeostasis. Mechanistically, TMBIM1 binds to and promotes the autoubiquitination and degradation of NEDD4, which is an E3 ligase that stabilizes PPARγ. Our data show that TMBIM1 is a potent repressor of adipogenesis and a potential therapeutic target for obesity-related metabolic disease.

4.
Molecules ; 26(4)2021 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-33671270

RESUMO

The aim of this work was to characterize biogenic amines (BAs) in different parts of Lycium barbarum L. using HPLC with dansyl chloride derivatization, and jointly, to provide referential data for further exploration and utilization of Lycium barbarum L. The linear correlation coefficients for all BAs were above 0.9989. The limits of detection and quantification were 0.015-0.075 and 0.05-0.25 µg/mL, respectively. The relative standard deviations for the intra-day and inter-day precision were 0.66-2.69% and 0.91-4.38%. The described method has good repeatability and intermediate precision for the quantitative determination of BAs in different parts of Lycium barbarum L. Satisfactory recovery for all amines was obtained (79.3-110.3%). The result showed that there were four kinds of BAs. The highest putrescine content (20.9 ± 3.2 mg/kg) was found in the flower. The highest histamine content (102.7 ± 5.8 mg/kg) was detected in the bark, and the highest spermidine (13.3 ± 1.6 mg/kg) and spermine (23.7 ± 2.0 mg/kg) contents were detected in the young leaves. The high histamine (HIS) content in the bark may be one of the reasons why all of the parts of Lycium barbarum L., except the bark, are used for medicine or food in China. Meanwhile, the issue of the high concentration of HIS should be considered when exploiting or utilizing the bark of Lycium barbarum L.


Assuntos
Aminas Biogênicas/análise , Cromatografia Líquida de Alta Pressão/métodos , Compostos de Dansil/química , Lycium/química , Espectrometria de Massas , Padrões de Referência , Reprodutibilidade dos Testes , Soluções
5.
Pharmacology ; 106(3-4): 189-201, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33621976

RESUMO

AIM: It has been reported that necrostatin-1 (Nec-1) is a specific necroptosis inhibitor that could attenuate programmed cell death induced by myocardial ischemia/reperfusion (I/R) injury. This study aimed to observe the effect and mechanism of novel Nec-1 analog (Z)-5-(3,5-dimethoxybenzyl)-2-imine-1-methylimidazolin-4-1 (DIMO) on myocardial I/R injury. METHODS: Male SD rats underwent I/R injury with or without different doses of DIMO (1, 2, or 4 mg/kg) treatment. Isolated neonatal rat cardiomyocytes were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) treatment with or without DIMO (0.1, 1, 10, or 100 µM). Myocardial infarction was measured by TTC staining. Cardiomyocyte injury was assessed by lactate dehydrogenase assay (LDH) and flow cytometry. Receptor-interacting protein 1 kinase (RIP1K) and autophagic markers were detected by co-immunoprecipitation and Western blotting analysis. Molecular docking of DIMO into the ATP binding site of RIP1K was performed using GLIDE. RESULTS: DIMO at doses of 1 or 2 mg/kg improved myocardial infarct size. However, the DIMO 4 mg/kg dose was ineffective. DIMO at the dose of 0.1 µM decreased LDH leakage and the ratio of PI-positive cells followed by OGD/R treatment. I/R or OGD/R increased RIP1K expression and in its interaction with RIP3K, as well as impaired myocardial autophagic flux evidenced by an increase in LC3-II/I ratio, upregulated P62 and Beclin-1, and activated cathepsin B and L. In contrast, DIMO treatment reduced myocardial cell death and reversed the above mentioned changes in RIP1K and autophagic flux caused by I/R and OGD/R. DIMO binds to RIP1K and inhibits RIP1K expression in a homology modeling and ligand docking. CONCLUSION: DIMO exerts cardioprotection against I/R- or OGD/R-induced injury, and its mechanisms may be associated with the reduction in RIP1K activation and restoration impaired autophagic flux.


Assuntos
Autofagia/efeitos dos fármacos , Cardiotônicos/farmacologia , Cardiotônicos/uso terapêutico , Imidazóis/química , Indóis/química , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Animais , Animais Recém-Nascidos , Proteína Beclina-1/metabolismo , Catepsina B/metabolismo , Catepsina L/metabolismo , Morte Celular/efeitos dos fármacos , Hemodinâmica/efeitos dos fármacos , Masculino , Proteínas Associadas aos Microtúbulos/metabolismo , Simulação de Acoplamento Molecular , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/prevenção & controle , Traumatismo por Reperfusão Miocárdica/metabolismo , Miócitos Cardíacos/efeitos dos fármacos , Necroptose/efeitos dos fármacos , Cultura Primária de Células , Proteínas Serina-Treonina Quinases/biossíntese , Proteínas Serina-Treonina Quinases/metabolismo , Ratos Sprague-Dawley , Proteína Serina-Treonina Quinases de Interação com Receptores/metabolismo , Homologia Estrutural de Proteína
6.
Zhongguo Dang Dai Er Ke Za Zhi ; 22(9): 1017-1026, 2020 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-32933637

RESUMO

OBJECTIVE: To study the effect of advanced maternal age (AMA) on the development of hippocampal neural stem cells in offspring rats. METHODS: Ten 3-month-old and ten 12-month-old female Sprague-Dawley rats were housed individually with 3-month-old male rats (1:1, n=20), whose offspring rats were assigned to a control group and an AMA group. A total of 40 rats were randomly selected from each group. Immunofluorescence assay and Western blot were used to localize and determine the levels of protein expression of Nestin and doublecortin (DCX) on day 7 as well as neuronal nuclear antigen (NeuN) and glial fibrillary acidic protein (GFAP) on day 28 (n=8 for each marker). Immunofluorescence assay was also used to localize the hippocampal expression of polysialylated isoforms of neural cell adhesion molecule (PSA-NCAM) on day 14 (n=8 for each marker). RESULTS: According to the Western blot results, the AMA group had significantly lower protein expression of DCX than the control group (P<0.05), while there were no significant differences in the protein expression of Nestin, NeuN, and GFAP between the two groups (P>0.05). According to the results of immunofluorescence assay, the AMA group had significantly lower protein expression of Nestin, DCX, and PSA-NCAM in the hippocampal dentate gyrus (DG) region than the control group (P<0.05), while there were no significant differences in the above indices in the hippocampal CA1 and CA3 regions between the two groups (P>0.05). The AMA group had significantly higher expression of NeuN in the hippocampal CA1 region than the control group (P<0.01), while there were no significant differences in the expression of NeuN in the hippocampal DG and CA3 regions between the two groups (P>0.05). The AMA group had significantly lower expression of GFAP in the hippocampal CA1, CA3, and DG regions than the control group (P<0.05). CONCLUSIONS: AMA may cause inhibition of proliferation, survival, and migration of hippocampal neural stem cells. AMA may also affect their differentiation into neurons and astrocytes, which will eventually lead to developmental disorders of hippocampal neural stem cells in offspring rats.


Assuntos
Células-Tronco Neurais , Animais , Feminino , Hipocampo , Masculino , Idade Materna , Neurônios , Ratos , Ratos Sprague-Dawley
7.
Aging Cell ; 19(10): e13224, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32866348

RESUMO

CD47 serves as a ligand for signaling regulatory protein α (SIRPα) and as a receptor for thrombospondin-1 (TSP-1). Although CD47, TSP-1, and SIRPα are thought to be involved in the clearance of aged red blood cells (RBCs), aging-associated changes in the expression and interaction of these molecules on RBCs have been elusive. Using direct stochastic optical reconstruction microscopy (dSTORM)-based imaging and quantitative analysis, we can report that CD47 molecules on young RBCs reside as nanoclusters with little binding to TSP-1, suggesting a minimal role for TSP-1/CD47 signaling in normal RBCs. On aged RBCs, CD47 molecules decreased in number but formed bigger and denser clusters, with increased ability to bind TSP-1. Exposure of aged RBCs to TSP-1 resulted in a further increase in the size of CD47 clusters via a lipid raft-dependent mechanism. Furthermore, CD47 cluster formation was dramatically inhibited on thbs1-/- mouse RBCs and associated with a significantly prolonged RBC lifespan. These results indicate that the strength of CD47 binding to its ligand TSP-1 is predominantly determined by the distribution pattern and not the amount of CD47 molecules on RBCs, and offer direct evidence for the role of TSP-1 in phagocytosis of aged RBCs. This study provides clear nanoscale pictures of aging-associated changes in CD47 distribution and TSP-1/CD47 interaction on the cell surface, and insights into the molecular basis for how these molecules coordinate to remove aged RBCs.


Assuntos
Envelhecimento/sangue , Antígeno CD47/sangue , Eritrócitos/metabolismo , Trombospondina 1/sangue , Animais , Camundongos , Camundongos Endogâmicos C57BL , Oligopeptídeos/sangue
8.
Sensors (Basel) ; 19(17)2019 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-31466246

RESUMO

Air pollution is one of the major threats to human health. The monitoring of toxic NO2 gas in urban air emission pollution is becoming increasingly important. Thus, the development of an NO2 sensor with low power consumption, low cost, and high performance is urgent. In this paper, a planar structural micro hotplate gas sensor based on an AlN ceramic substrate with an annular Pt film heater was designed and prepared by micro-electro-mechanical system (MEMS) technology, in which Pt/Nb/In2O3 composite semiconductor oxide was used as the sensitive material with a molar ratio of In:Nb = 9:1. The annular thermal isolation groove was designed around the heater to reduce the power consumption and improve the thermal response rate. Furthermore, the finite element simulation analysis of the thermal isolation structure of the sensor was carried out by using ANSYS software. The results show that a low temperature of 94 °C, low power consumption of 150 mW, and low concentration detection of 1 to 10 ppm NO2 were simultaneously realized for the Nb-doped In2O3-based gas sensor. Our findings provide a promising strategy for the application of In2O3-based sensors in highly effective and low concentration NO2 detection.

9.
BMC Genomics ; 20(1): 587, 2019 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-31315571

RESUMO

BACKGROUND: Atrazine is widely used in agriculture and is a known endocrine disrupting chemical. Atrazine can seep into the water body through surface, posing a potential threat to the aquatic ecological environment and human drinking water source. In vertebrate, studies have shown that it can affect reproduction and development seriously, but its molecular mechanism for aquatic animals is unknown. Aquaculture is very common in China, especially common carp, whose females grow faster than males. However, the effects of atrazine on the reproduction of carp, especially miRNA, have not been investigated. RESULTS: In this study, common carp (Cyprinus carpio) at two key developmental stages were exposed to atrazine in vitro. Sex ratio was observed to analyze the effect of atrazine on the sex. MiRNA expression profiles were analysed to identify miRNAs related to gonad development and to reveal the atrazine mechanisms interfering with gonad differentiation. The results showed that the sex ratio was biased towards females. Atrazine exposure caused significant alteration of multiple miRNAs. Predicted targets of differently-expressed miRNAs were involved in many reproductive biology signalling pathways. CONCLUSIONS: Our results indicate that atrazine promoted the expression of female-biased genes by decreasing miRNAs in primordial gonad. In addition, our results indicate that atrazine can up-regulate aromatase expression through miRNAs, which supports the hypothesis that atrazine has endocrine-disrupting activity by altering the gene expression profile of the Hypothalamus-Pituitary-Gonad axis through its corresponding miRNAs.


Assuntos
Atrazina/toxicidade , Carpas/genética , Disruptores Endócrinos/toxicidade , Gônadas/efeitos dos fármacos , MicroRNAs/genética , Diferenciação Sexual/efeitos dos fármacos , Animais , Aromatase/genética , Feminino , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Gônadas/embriologia , Masculino , Razão de Masculinidade , Transcriptoma/efeitos dos fármacos
10.
Sci Rep ; 9(1): 8691, 2019 06 18.
Artigo em Inglês | MEDLINE | ID: mdl-31213623

RESUMO

Yellow River carp is widely cultivated in the world due to its economic value in aquaculture, and the faster growth of females compared to males. It is believed that microRNAs (miRNA) are involved in gonadal differentiation and development. qPCR is the most preferred method for miRNA functional analysis. Reliable reference genes for normalization in qRT-PCR are the key to ensuring the accuracy of this method. The aim of present research was to evaluate as well as identify the efficacy of reference genes for miRNA expression using qRT-PCR in Yellow River carp. Nine ncRNAs (miR-101, miR-23a, let7a, miR-26a, miR-146a, miR-451, U6, 5S, and 18S) were chosen and tested in four sample sets: (1) different tissues in adult carp, (2) different tissues in juvenile carp, (3) different early developmental stages of carp, and (4) different developmental stages of carp gonads. The stability and suitability values were calculated using NormFinder, geNorm, and BestKeeper software. The results showed that 5S was a suitable reference gene in different tissues of adult and juvenile carp. The genes 5S, 18S, and U6 were the most stable reference genes in the early developmental stages of carp. Let-7a and miR-23a were considered as the suitable reference genes in the development of gonads. All these reference genes were subsequently validated using miR-430. The results showed that genes 5S and 18S were the most suitable reference genes to normalize miRNA expression under normal growth conditions in early different developmental stages. The genes Let-7a, and miR-23a were the most suitable in different developmental stages. The present study is the first comprehensive study of the stability of miRNA reference genes in Yellow River carp, providing valuable as well as basic data for investigating more accurate miRNA expression during gonadal differentiation and development of carp.


Assuntos
Carpas/genética , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica no Desenvolvimento , MicroRNAs/genética , Animais , Carpas/crescimento & desenvolvimento , Feminino , Perfilação da Expressão Gênica/normas , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Masculino , Padrões de Referência , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rios
11.
J Appl Toxicol ; 38(11): 1437-1446, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30051583

RESUMO

Cadmium is considered one of the most harmful carcinogenic heavy metals in the human body. Although many scientists have performed research on cadmium toxicity mechanism, the toxicokinetic process of cadmium toxicity remains unclear. In the present study, the kinetic response of proteome in/and A549 cells to exposure of exogenous cadmium was profiled. A549 cells were treated with cadmium sulfate (CdSO4 ) for different periods and expressions of proteins in cells were detected by two-dimensional gel electrophoresis. The kinetic expressions of proteins related to cadmium toxicity were further investigated by reverse transcription-polymerase chain reaction and western blotting. Intracellular cadmium accumulation and content fluctuation of several essential metals were observed after 0-24 hours of exposure by inductively coupled plasma mass spectrometry. Fifty-four protein spots showed significantly differential responses to CdSO4 exposure at both 4.5 and 24 hours. From these proteins, four expression patterns were concluded. Their expressions always exhibited a maximum abundance ratio after CdSO4 exposure for 24 hours. The expression of metallothionein-1 and ZIP-8, concentration of total protein, and contents of cadmium, zinc, copper, cobalt and manganese in cells also showed regular change. In synthesis, the replacement of the essential metals, the inhibition of the expression of metal storing protein and the activation of metal efflux system are involved in cadmium toxicity.


Assuntos
Cádmio/toxicidade , Proteínas de Transporte de Cátions/metabolismo , Metalotioneína/metabolismo , Proteoma/metabolismo , Células A549 , Cádmio/metabolismo , Proteínas de Transporte de Cátions/genética , Técnicas de Cultura de Células , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Expressão Gênica/efeitos dos fármacos , Humanos , Metalotioneína/genética , Proteoma/genética , Fatores de Tempo , Toxicocinética
12.
Artigo em Inglês | MEDLINE | ID: mdl-29856688

RESUMO

Benzo[α]pyrene (BaP) is a well-known carcinogen in edible oil. In this study, a method combined solid-phase extraction (SPE) with fluorescent detection was developed using tetraoxocalix[2]arene[2]triazine sorbent (SiO2-OCA) for the clean-up and enrichment of BaP. The interaction between SiO2-OCA and BaP involves a donor-acceptor complex mechanism. The experimental procedure was as follows: BaP was extracted from edible oil with DMF/H2O (9:1, v/v). Then, the ratio of DMF/H2O was adjusted to 1:2 prior to SPE. The final concentrate was analysed using a fluorescence detector at excitation and emission wavelengths of 255 and 420 nm. The method was fully validated. The linearity was in the range of 0.1-100 µg kg-1 with a coefficient of 0.999. The limits of detection and quantification were 0.03 and 0.1 µg kg-1, respectively. The average recoveries were in the range of 88.0 - 122.3%. The intraday and interday precisions were 6.8% and 9.2%, respectively. Compared with other methods, the method reported in this article shows a good detection limit, high reproducibility and recovery and linearity over a broad concentration range. This established method was also applied to evaluate real samples. The concentration of six tested samples was below 5 µg kg-1.


Assuntos
Benzo(a)pireno/análise , Calixarenos/química , Óleos Vegetais/química , Dióxido de Silício/química , Extração em Fase Sólida , Triazenos/química
13.
Zhonghua Nan Ke Xue ; 24(12): 1064-1068, 2018 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-32212483

RESUMO

Objective: To evaluate the effect of L-carnitine (LC) on low sperm acrosin activity in infertile man. METHODS: A total of 240 male infertility patients with low sperm acrosin activity were randomly assigned to an LC group (n = 180) and a control group (n = 60) to be treated with LC (1g, tid) and vitamin E (VE) capsules (100 mg, tid) respectively, both for 3 months. Based on the results of routine semen analysis, the patients in the experimental group were further divided into oligozoospermia, asthenozoospermia and normozoospermia subgroups. Semen parameters and sperm acrosin activity were examined before and after treatment. RESULTS: Totally, 220 of the patients completed the treatment and follow-up, 163 in the LC medication and 57 in the VE control group. Compared with the baseline, the percentage of progressively motile sperm (PMS) was significantly increased in the LC group after 3 months of treatment (ï¼»32.58 ± 1.13ï¼½% vs ï¼»36.35 ± 1.26ï¼½%, P < 0.05), and so was sperm acrosin activity (ï¼»37.05±0.66ï¼½ vs ï¼»58.61±1.93ï¼½ µIU/106 sperm, P < 0.01). Sperm concentration, PMS and sperm acrosin activity were also improved in the VE control group after treatment, but with no statistically significant difference (P > 0.05). In comparison with pretreatment, remarkable increases were observed after LC medication in sperm concentration in the oligozoospermia subgroup (ï¼»11.27 ± 0.73ï¼½ vs ï¼»21.82 ± 4.21ï¼½ ×106/ml, P < 0.01) and PMS in the asthenozoospermia patients (ï¼»20.61 ± 0.85ï¼½% vs ï¼»29.81 ± 1.88ï¼½%, P < 0.01). And sperm acrosin activity was even higher after treatment in the asthenozoospermia than in the oligozoospermia and normozoospermia subgroups (ï¼»60.85 ± 3.04ï¼½ vs ï¼»56.32 ± 2.86ï¼½ and ï¼»57.09 ± 6.31ï¼½ µIU/106 sperm, P < 0.05). CONCLUSIONS: L-carnitine can effectively elevate sperm acrosin activity in male infertility patients, particularly in those with asthenozoospermia.


Assuntos
Carnitina , Infertilidade Masculina , Motilidade Espermática , Acrosina , Carnitina/farmacologia , Carnitina/uso terapêutico , Humanos , Infertilidade Masculina/tratamento farmacológico , Masculino , Sêmen , Contagem de Espermatozoides , Motilidade Espermática/efeitos dos fármacos , Espermatozoides , Vitamina E/uso terapêutico , Vitaminas/uso terapêutico
14.
Cell Rep ; 15(11): 2387-99, 2016 06 14.
Artigo em Inglês | MEDLINE | ID: mdl-27264185

RESUMO

The synchronized activity of six layers of cortical neurons is critical for sensory perception and the control of voluntary behavior, but little is known about the synaptic mechanisms of cortical synchrony across layers in behaving animals. We made single and dual whole-cell recordings from the primary somatosensory forepaw cortex in awake mice and show that L2/3 and L5 excitatory neurons have layer-specific intrinsic properties and membrane potential dynamics that shape laminar-specific firing rates and subthreshold synchrony. First, while sensory and movement-evoked synaptic input was tightly correlated across layers, spontaneous action potentials and slow spontaneous subthreshold fluctuations had laminar-specific timing; second, longer duration forepaw movement was associated with a decorrelation of subthreshold activity; third, spontaneous and sensory-evoked forepaw movements were signaled more strongly by L5 than L2/3 neurons. Together, our data suggest that the degree of translaminar synchrony is dependent upon the origin (sensory, spontaneous, and movement) of the synaptic input.


Assuntos
Comportamento Animal/fisiologia , Potenciais da Membrana/fisiologia , Córtex Somatossensorial/fisiologia , Animais , Extremidades/fisiologia , Camundongos , Movimento/fisiologia , Estimulação Física , Células Piramidais/fisiologia , Córtex Somatossensorial/citologia , Sinapses/fisiologia , Tato , Vigília
15.
Biol Trace Elem Res ; 170(1): 75-83, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26198133

RESUMO

Intracellular metal elements exist in mammalian cells with the concentration range from picomoles per litre to micromoles per litre and play a considerable role in various biological procedures. Element provided by different species can influence the availability and distribution of the element in a cell and could lead to different biological effects on the cell's growth and function. Zinc as an abundant and widely distributed essential trace element, is involved in numerous and relevant physiological functions. Zinc homeostasis in cells, which is regulated by metallothioneins, zinc transporter/SLC30A, Zrt-/Irt-like proteins/SLC39A and metal-response element-binding transcription factor-1 (MTF-1), is crucial for normal cellular functioning. In this study, we investigated the influences of different zinc species, zinc sulphate, zinc gluconate and bacitracin zinc, which represented inorganic, organic and biological zinc species, respectively, on cell cycle, viability and apoptosis in MDAMB231 cells. It was found that the responses of cell cycle, apoptosis and death to different zinc species in MDAMB231 cells are different. Western blot analysis of the expression of several key proteins in regulating zinc-related transcription, cell cycle, apoptosis, including MTF-1, cyclin B1, cyclin D1, caspase-8 and caspase-9 in treated cells further confirmed the observed results on cell level.


Assuntos
Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Zinco/farmacologia , Compartimento Celular , Linhagem Celular Tumoral , Humanos , Microscopia Confocal , Zinco/metabolismo
16.
PLoS One ; 10(7): e0133451, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26196515

RESUMO

Zinc, an essential trace element, is involved in many important physiological processes. Cell responses to zinc stress show time-dependent effects besides concentration-dependence and tissue-specificity. Herein, we investigated the time-dependent differential expression of the proteome in A549 cells after administered with ZnSO4 for both 9 and 24 h using 2DE. 123 differentially expressed protein spots were detected, most of which were up-regulated by Zn2+ treatment. Interestingly, 49 proteins exhibited significant differential expression repeatedly during these two treatment periods, and moreover showed a conserved change with different ratios and four time-dependent expression patterns. Pattern 1 (up-regulated with rapid initial induction and subsequent repression) and pattern 4 (down-regulated with steady repression) were the predominant expression patterns. The abundances of the proteins in patterns 1 and 4 after 24 h of zinc treatment are always lower than that after 9 h, indicating that exogenous zinc reduced the expression of proteins in cells after 24 h or longer. Importantly, these findings could also reflect the central challenge in detecting zinc homeostasis proteins by 2DE or other high throughput analytical methods resulting from slight variation in protein expression after certain durations of exogenous zinc treatment and/or low inherent protein content in cells. These time-dependent proteome expression patterns were further validated by measuring dynamic changes in protein content in cells and in expression of two proteins using the Bradford method and western blotting, respectively. The time-dependent changes in total zinc and free Zn2+ ion contents in cells were measured using ICP-MS and confocal microscopy, respectively. The kinetic process of zinc homeostasis regulated by muffling was further revealed. In addition, we identified 50 differentially expressed proteins which are predominantly involved in metabolic process, cellular process or developmental process, and function as binding, catalytic activity or structural molecule activity. This study further elucidates our understanding of dynamic nature of the cellular response to zinc stress and the mechanism of zinc homeostasis.


Assuntos
Proteoma/análise , Proteômica/métodos , Estresse Fisiológico , Sulfato de Zinco/farmacologia , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Western Blotting , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Eletroforese em Gel Bidimensional , Humanos , Cinética , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Espectrometria de Massas/métodos , Microscopia Confocal , Proteoma/classificação , Proteoma/metabolismo , Fatores de Tempo , Regulação para Cima/efeitos dos fármacos
17.
Toxicology ; 333: 76-88, 2015 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-25896364

RESUMO

The most well-known cause of chemotherapy-induced cardiotoxicity is doxorubicin (DOX). The ubiquitin-proteasome system (UPS) is the main cellular machinery for protein degradation in eukaryotic cells. However, the expression pattern of the UPS in DOX-induced cardiotoxicity remains unclear. C57BL/6 mice were intraperitoneally injected with a single dose of DOX (15mg/kg). After 1, 3 and 5 days, cardiac function and apoptosis were detected with echocardiography and TUNEL assay. Microarray assay and qPCR analysis were also performed at day 5. We found that DOX caused a significant decrease in cardiac function at day 5 and increase in cardiomyocyte apoptosis at days 3 and 5. Microarray data revealed that totally 1185 genes were significantly regulated in DOX-treated heart, and genes involved in apoptosis and the UPS were mostly altered. Among them, the expression of 3 immunoproteasome catalytic subunits (ß1i, ß2i and ß5i) was markedly down-regulated. Moreover, DOX significantly decreased proteasome activities and enhanced polyubiquitinated proteins in the heart. Importantly, overexpression of immunoproteasome catalytic subunits (ß1i, ß2i or ß5i) significantly attenuated DOX-induced cardiomyocyte apoptosis and other UPS gene expression while knockdown of them significantly increased DOX-induced cardiomyocyte apoptosis. These effects were partially associated with increased degradation of multiple pro-apoptotic proteins. In conclusion, our studies suggest that immunoproteasome plays an important role in DOX-induced cardiomyocyte apoptosis, and may be a novel therapeutic target for prevention of DOX-induced cardiotoxicity.


Assuntos
Antibióticos Antineoplásicos , Doxorrubicina , Perfilação da Expressão Gênica , Cardiopatias/genética , Miócitos Cardíacos/enzimologia , Complexo de Endopeptidases do Proteassoma/genética , Animais , Apoptose , Células Cultivadas , Modelos Animais de Doenças , Feminino , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica , Cardiopatias/induzido quimicamente , Cardiopatias/enzimologia , Cardiopatias/imunologia , Cardiopatias/patologia , Cardiopatias/fisiopatologia , Camundongos Endogâmicos C57BL , Miócitos Cardíacos/imunologia , Análise de Sequência com Séries de Oligonucleotídeos , Complexo de Endopeptidases do Proteassoma/metabolismo , Interferência de RNA , Fatores de Tempo , Transcrição Genética , Transfecção , Ubiquitinação
18.
Cardiovasc Toxicol ; 15(4): 377-93, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25575753

RESUMO

Clinical use of doxorubicin (DOX) in cancer therapy is limited by its dose-dependent cardiotoxicity. But molecular mechanisms underlying this phenomenon have not been well defined. This study was to investigate the effect of DOX on the changes of global genomics in hearts. Acute cardiotoxicity was induced by giving C57BL/6J mice a single intraperitoneal injection of DOX (15 mg/kg). Cardiac function and apoptosis were monitored using echocardiography and TUNEL assay at days 1, 3 and 5. Myocardial glucose and ATP levels were measured. Microarray assays were used to screen gene expression profiles in the hearts at day 5, and the results were confirmed with qPCR analysis. DOX administration caused decreased cardiac function, increased cardiomyocyte apoptosis and decreased glucose and ATP levels. Microarrays showed 747 up-regulated genes and 438 down-regulated genes involved in seven main functional categories. Among them, metabolic pathway was the most affected by DOX. Several key genes, including 2,3-bisphosphoglycerate mutase (Bpgm), hexokinase 2, pyruvate dehydrogenase kinase, isoenzyme 4 and fructose-2,6-bisphosphate 2-phosphatase, are closely related to glucose metabolism. Gene co-expression networks suggested the core role of Bpgm in DOX cardiomyopathy. These results obtained in mice were further confirmed in cultured cardiomyocytes. In conclusion, genes involved in glucose metabolism, especially Bpgm, may play a central role in the pathogenesis of DOX-induced cardiotoxicity.


Assuntos
Antibióticos Antineoplásicos , Cardiomiopatias/genética , Doxorrubicina , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica/efeitos dos fármacos , Redes Reguladoras de Genes/efeitos dos fármacos , Análise de Sequência com Séries de Oligonucleotídeos , Trifosfato de Adenosina/metabolismo , Animais , Apoptose/genética , Bisfosfoglicerato Mutase/genética , Bisfosfoglicerato Mutase/metabolismo , Cardiomiopatias/induzido quimicamente , Cardiomiopatias/patologia , Cardiomiopatias/fisiopatologia , Células Cultivadas , Modelos Animais de Doenças , Metabolismo Energético/genética , Glucose/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , Ratos Sprague-Dawley , Fatores de Tempo
19.
Nat Neurosci ; 17(11): 1560-6, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25262494

RESUMO

The temperature of an object provides important somatosensory information for animals performing tactile tasks. Humans can perceive skin cooling of less than one degree, but the sensory afferents and central circuits that they engage to enable the perception of surface temperature are poorly understood. To address these questions, we examined the perception of glabrous skin cooling in mice. We found that mice were also capable of perceiving small amplitude skin cooling and that primary somatosensory (S1) cortical neurons were required for cooling perception. Moreover, the absence of the menthol-gated transient receptor potential melastatin 8 ion channel in sensory afferent fibers eliminated the ability to perceive cold and the corresponding activation of S1 neurons. Our results identify parts of a neural circuit underlying cold perception in mice and provide a new model system for the analysis of thermal processing and perception and multimodal integration.


Assuntos
Neurônios/fisiologia , Percepção/fisiologia , Sensação Térmica/fisiologia , Animais , Temperatura Baixa , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Estimulação Física/métodos , Pele/inervação , Fenômenos Fisiológicos da Pele , Canais de Cátion TRPM/metabolismo , Tato/fisiologia
20.
PLoS One ; 9(8): e105797, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25162517

RESUMO

Zinc plays a critical role in many biological processes. However, it is toxic at high concentrations and its homeostasis is strictly regulated by metal-responsive transcription factor 1 (MTF-1) together with many other proteins to protect cells against metal toxicity and oxidative stresses. In this paper, we used high-resolution two-dimensional gel electrophoresis (2DE) to profile global changes of the whole soluble proteome in human lung adenocarcinoma (A549) cells in response to exogenous zinc treatment for 24 h. Eighteen differentially expressed proteins were identified by MALDI TOF/TOF and MASCOT search. In addition, we used Western blotting and RT-PCR to examine the time-dependent changes in expression of proteins regulated by MTF-1 in response to Zn treatment, including the metal binding protein MT-1, the zinc efflux protein ZnT-1, and the zinc influx regulator ZIP-1. The results indicated that variations in their mRNA and protein levels were consistent with their functions in maintaining the homeostasis of zinc. However, the accumulation of ZIP-1 transcripts was down-regulated while the protein level was up-regulated during the same time period. This may be due to the complex regulatory mechanism of ZIP-1, which is involved in multiple signaling pathways. Maximal changes in protein abundance were observed at 10 h following Zn treatment, but only slight changes in protein or mRNA levels were observed at 24 h, which was the time-point frequently used for 2DE analyses. Therefore, further study of the time-dependent Zn-response of A549 cells would help to understand the dynamic nature of the cellular response to Zn stress. Our findings provide the basis for further study into zinc-regulated cellular signaling pathways.


Assuntos
Proteínas de Ligação a DNA/genética , Células Epiteliais/efeitos dos fármacos , Proteoma/genética , Alvéolos Pulmonares/efeitos dos fármacos , RNA Mensageiro/genética , Fatores de Transcrição/genética , Sulfato de Zinco/farmacologia , Proteínas de Transporte de Cátions/genética , Proteínas de Transporte de Cátions/metabolismo , Linhagem Celular Tumoral , Proteínas de Ligação a DNA/metabolismo , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Homeostase/genética , Humanos , Metalotioneína/genética , Metalotioneína/metabolismo , Proteoma/metabolismo , Alvéolos Pulmonares/citologia , Alvéolos Pulmonares/metabolismo , RNA Mensageiro/metabolismo , Transdução de Sinais , Estresse Fisiológico , Fatores de Tempo , Fatores de Transcrição/metabolismo
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