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1.
Food Chem ; 458: 140266, 2024 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-38964095

RESUMO

The interaction between proteins and soluble dietary fibers plays a vital role in the development of animal-derived foods. Herein, the effects of different contents (0-3.0%) of round-bracted psyllium husk powder (PHP) on the gelation behavior, microstructure, and intermolecular interactions of Andrias davidianus myofibrillar protein (MP) were investigated. Rheological and chemical forces suggested that PHP (1.5%-2.0%) enhanced the functional properties of MP at low ionic strength, thereby increasing the viscoelasticity of mixed gels. SDS-PAGE revealed that PHP reinforced the cross-linking and aggregation of protein molecules. Circular dichroism spectroscopy, low-field nuclear magnetic resonance, and scanning electron microscopy demonstrated that PHP induced the transformation of α-helix (decreased by 14.85%) to an ordered ß-sheet structure (increased by 81.58%), which was more favorable for the formation of dense network structure and improved (10.53%) the water retention of MP gels. This study provided new insights for PHP to effectively meliorate the heat-induced gelling properties of MP.

2.
Neuroscience ; 526: 196-203, 2023 08 21.
Artigo em Inglês | MEDLINE | ID: mdl-37419407

RESUMO

Tau protein hyperphosphorylation and formation of intracellular neurofibrillary tangles (NFTs) are one of the histopathological hallmarks of Alzheimer's disease (AD) and positively correlated with the severity of AD symptoms. NFTs contain a large number of metal ions that play an important role in regulating tau protein phosphorylation and AD progression. Extracellular tau induces primary phagocytosis of stressed neurons and neuronal loss by activating microglia. Here, we studied the effects of a multi-metal ion chelator, DpdtpA, on tau-induced microglial activation and inflammatory responses and the underlying mechanisms. Treatment with DpdtpA attenuated the increase in the expression of NF-κB and production of inflammatory cytokines, IL-1ß, IL-6 and IL-10, in rat microglial cells induced by expression of human tau40 proteins. Treatment with DpdtpA also suppressed tau protein expression and phosphorylation. Moreover, treatment with DpdtpA prevented tau-induced activation of glycogen synthase kinase-3ß (GSK-3ß) and inhibition of phosphatidylinositol-3-hydroxy kinase (PI3K)/AKT. Collectively, these results show that DpdtpA can attenuate tau phosphorylation and inflammatory responses of microglia by regulating the PI3K/AKT/GSK-3ß signal pathways, providing a new option to alleviate neuroinflammation for the treatment of AD.


Assuntos
Doença de Alzheimer , Proteínas tau , Ratos , Humanos , Animais , Proteínas tau/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Fosfatidilinositol 3-Quinase/farmacologia , Fosforilação , Microglia/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , Transdução de Sinais , Doença de Alzheimer/metabolismo , Quelantes/farmacologia
3.
Med Res Rev ; 43(5): 1346-1373, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-36924449

RESUMO

The P2X7 receptor is an exceptional member of the P2X purinergic receptor family, with its activation requiring high concentrations of extracellular adenosine 5'-triphosphate (ATP) that are often associated with tissue damage and inflammation. In the central nervous system (CNS), it is highly expressed in glial cells, particularly in microglia. In this review, we discuss the role and mechanisms of the P2X7 receptor in mediating neuroinflammation and other pathogenic events in a variety of traumatic CNS damage conditions, which lead to loss of neurological and cognitive functions. We raise the perspective on the steady progress in developing CNS-penetrant P2X7 receptor-specific antagonists that leverage the ATP-P2X7 receptor signaling axis as a potential therapeutic strategy to alleviate traumatic CNS damage and related complications.


Assuntos
Sistema Nervoso Central , Receptores Purinérgicos P2X7 , Humanos , Microglia , Antagonistas do Receptor Purinérgico P2X/farmacologia , Antagonistas do Receptor Purinérgico P2X/uso terapêutico , Trifosfato de Adenosina
4.
Brain Sci ; 12(9)2022 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-36138981

RESUMO

Gasdermin D-executing pyroptosis mediated by NLRP3 inflammasomes has been recognized as a key pathogenesis during stroke. Hydrogen sulfide (H2S) could protect CNS against ischemia/reperfusion (I/R)-induced neuroinflammation, while the underlying mechanism remains unclear. The study applied the middle cerebral artery occlusion/reperfusion (MCAO/R) model to investigate how the brain and the retinal injuries were alleviated in sodium hydrogen sulfide (NaHS)-treated rats. The rats were assigned to four groups and received an intraperitoneal injection of 50 µmol/kg NaHS or NaCl 15 min after surgery. Neurological deficits were evaluated using the modified neurologic severity score. The quantification of pro-inflammatory cytokines, NLRP3, caspase-1, and GSDMD were determined by ELISA and Western blot. Cortical and retinal neurodegeneration and cell pyroptosis were determined by histopathologic examination. Results showed that NaHS rescued post-stroke neurological deficits and infarct progression, improved retina injury, and attenuated neuroinflammation in the brain cortexes and the retinae. NaHS administration inhibits inflammation by blocking the NLRP3/caspase-1/GSDMD pathway and further suppressing neuronal pyroptosis. This is supported by the fact that it reversed the high-level of NLRP3, caspase-1, and GSDMD following I/R. Our findings suggest that compounds with the ability to donate H2S could constitute a novel therapeutic strategy for ischemic stroke.

5.
Luminescence ; 34(2): 153-161, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30628166

RESUMO

Two new twinborn benzimidazole derivates (L and A), which bonded pyridine via the ester space on the opposite and adjacent positions of the benzene ring of benzimidazole respectively, were designed and synthesized. Compound L displayed fluorescence quenching response only towards copper(II) ions (Cu2+ ) in acetonitrile solution with high selectivity and sensitivity. However, compound A presented 'on-off' fluorescence response towards a wide range of metal ions to different degrees and did not have selectivity. Furthermore, compound L formed a 1:1 complex with Cu2+ and the binding constant between sensor L and Cu2+ was high at 6.02 × 104  M-1 . Job's plot, mass spectra, IR spectra, 1 H-NMR titration and density functional theory (DFT) calculations demonstrated the formation of a 1:1 complex between L and Cu2+ . Chemosensor L displayed a low limit of detection (3.05 × 10-6  M) and fast response time (15 s) to Cu2+ . The Stern-Volmer analysis illustrated that the fluorescence quenching agreed with the static quenching mode. In addition, the obvious difference of L within HepG2 cells in the presence and absence of Cu2+ indicated L had the recognition capability for Cu2+ in living cells.


Assuntos
Benzimidazóis/química , Cobre/análise , Corantes Fluorescentes/química , Imagem Óptica , Teoria da Densidade Funcional , Células Hep G2 , Humanos , Íons/análise , Espectrometria de Fluorescência
6.
Sci China C Life Sci ; 50(6): 731-8, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18026860

RESUMO

Plasma membrane (PM) proteome is one of the major subproteomes present in the cell, and is very important in liver function. In the present work, C57 mouse liver PM was purified by density-gradient centrifugation. The purified PM was verified by electron microscope analysis and Western blotting. The results showed that the PM was enriched by more than 20-fold and the contamination of mitochondria was reduced by 2-fold compared with the homogenization fraction. Proteins were separated by 2DE and 1DE, trypsin-digested and submitted to ESI-Q-TOF and MALDI-TOF-TOF mass spectrometry or directly digested in solution and analyzed by LC-ESI ion trap mass spectrometry. In all, 547 non-redundant mouse liver PM proteins were identified, of which 34% contributed to plasma membrane or plasma membrane-related proteins. This study optimized and evaluated the HLPP plasma membrane proteome analysis method and made a systematic analysis on PM proteome.


Assuntos
Membrana Celular/química , Extratos Hepáticos/análise , Fígado/química , Proteoma/análise , Animais , Fracionamento Celular , Extratos Hepáticos/química , Proteínas de Membrana/análise , Proteínas de Membrana/isolamento & purificação , Camundongos , Camundongos Endogâmicos C57BL , Proteoma/química
7.
J Proteome Res ; 6(1): 34-43, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17203946

RESUMO

Plasma membrane (PM) has very important roles in cell-cell interaction and signal transduction, and it has been extensively targeted for drug design. A major prerequisite for the analysis of PM proteome is the preparation of PM with high purity. Density gradient centrifugation has been commonly employed to isolate PM, but it often occurred with contamination of internal membrane. Here we describe a method for plasma membrane purification using second antibody superparamagnetic beads that combines subcellular fractionation and immunoisolation strategies. Four methods of immunoaffinity were compared, and the variation of crude plasma membrane (CPM), superparamagnetic beads, and antibodies was studied. The optimized method and the number of CPM, beads, and antibodies suitable for proteome analysis were obtained. The PM of mouse liver was enriched 3-fold in comparison with the density gradient centrifugation method, and contamination from mitochondria was reduced 2-fold. The PM protein bands were extracted and trypsin-digested, and the resulting peptides were resolved and characterized by MALDI-TOF-TOF and ESI-Q-TOF, respectively. Mascot software was used to analyze the data against IPI-mouse protein database. Nonredundant proteins (248) were identified, of which 67% are PM or PM-related proteins. No endoplasmic reticulum (ER) or nuclear proteins were identified according to the GO annotation in the optimized method. Our protocol represents a simple, economic, and reproducible tool for the proteomic characterization of liver plasma membrane.


Assuntos
Fracionamento Celular/métodos , Membrana Celular/metabolismo , Separação Imunomagnética/métodos , Proteômica/métodos , Animais , Comunicação Celular , Centrifugação com Gradiente de Concentração , Bases de Dados de Proteínas , Fígado/metabolismo , Camundongos , Mitocôndrias/metabolismo , Transdução de Sinais , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Tripsina/farmacologia
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