RESUMO
Pigs have anatomical and physiological characteristics comparable to those in humans and, therefore, are a favorable model for immune function research. Interferons (IFNs) and inflammasomes have essential roles in the innate immune system. Here, we report that G10, a human-specific agonist of stimulator of interferon genes (STING), activates both type I IFN and the canonical NLRP3 inflammasome in a STING-dependent manner in porcine cells. Without a priming signal, G10 alone transcriptionally stimulated Sp1-dependent p65 expression, thus triggering activation of the nuclear factor-κB (NF-κB) signaling pathway and thereby priming inflammasome activation. G10 was also found to induce potassium efflux- and NLRP3/ASC/Caspase-1-dependent secretion of IL-1ß and IL-18. Pharmacological and genetic inhibition of NLRP3 inflammasomes increased G10-induced type I IFN expression, thereby preventing virus infection, suggesting negative regulation of the NLRP3 inflammasome in the IFN response in the context of STING-mediated innate immune activation. Overall, our findings reveal a new mechanism through which G10 activates the NLRP3 inflammasome in porcine cells and provide new insights into STING-mediated innate immunity in pigs compared with humans.
Assuntos
Imunidade Inata/efeitos dos fármacos , Inflamassomos/agonistas , Interferon Tipo I/metabolismo , Proteínas de Membrana/agonistas , Proteína 3 que Contém Domínio de Pirina da Família NLR/agonistas , Tiazinas/farmacologia , Animais , Proteínas Adaptadoras de Sinalização CARD/metabolismo , Caspase 1/metabolismo , Chlorocebus aethiops , Células HEK293 , Humanos , Inflamassomos/genética , Inflamassomos/imunologia , Inflamassomos/metabolismo , Interferon Tipo I/genética , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/imunologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Transdução de Sinais , Fator de Transcrição Sp1/genética , Fator de Transcrição Sp1/metabolismo , Sus scrofa , Células THP-1 , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo , Células VeroRESUMO
Polydopamine (PDA) can be formed by monomeric self-polymerization in water. This convenient behavior was exploited to prepare a molecularly imprinted polymer (MIP) layer on the surface of multi-walled carbon nanotubes (MWCNTs) with sunset yellow (SY) as a template molecule. The prepared nanocomposites were characterized, and their electrochemical behavior towards SY was investigated. Under the optimized conditions, a glassy carbon electrode modified with the imprinted nanocomposite showed a highly selective and ultrasensitive electrochemical response to SY compared with the performance of control electrodes and previously reported electrochemical sensors for SY. The improved behavior of the developed sensor can be attributed to its superficial highly matched imprinted cavities on the excellent electrocatalytic matrix of MWCNTs and the electronic barrier of the non-imprinted PDA to outside molecules. The fabricated sensor expressed a linear relationship to SY concentrations from 2.2nM to 4.64µM with a detection limit of 1.4nM (S/N = 3). The sensor also exhibited excellent selectivity for SY over its structural analogs, good stability, and adequate reproducibility. The prepared sensor was successfully used to detect SY in real spiked samples. This methodology has potential application value and may be readily adapted to design other PDA-based MIP sensors.
Assuntos
Compostos Azo/análise , Corantes/análise , Técnicas Eletroquímicas/métodos , Indóis/química , Impressão Molecular/métodos , Nanotubos de Carbono/química , Polímeros/química , Análise de Alimentos/métodos , Limite de Detecção , Reprodutibilidade dos TestesRESUMO
INTRODUCTION: Magnetic Fe3O4 nanoparticles were prepared by coprecipitation and then were coated with SiO2 on the surface. MATERIALS AND METHODS: Fe3O4@SiO2 composite microspheres were modified by KH570. Using molecular imprinting technology, atrazine magnetic molecularly imprinted polymer was prepared by using atrazine as template molecule, methacrylic acid as functional monomer and ethylene glycol dimethacrylate as cross-linkers. The morphology, composition and magnetic properties of magnetic nanoparticles were characterized. The recognition selectivity of polymer was studied for template molecule and simulation by UV spectrophotometry. The adsorption properties and selectivity ability were analyzed by Scatchard analysis. RESULTS: Scatchard linear regression analysis indicated that there are two binding sites of the target molecules. The magnetic molecularly imprinted polymer has been applied to the analysis of atrazine in real samples. CONCLUSION: The results show that: the recovery rates and the relative standard deviation were 94.0â¼98.7% and 2.1â¼4.0% in corn, the recovery rates and the relative standard deviation were 88.7â¼93.5% and 2.8â¼7.2% in water.
