RÉSUMÉ
Activated macrophages play an important role in many inflammatory diseases including septic shock and atherosclerosis. However, the molecular mechanisms limiting macrophage activation are not completely understood. Members of the tripartite motif (TRIM) family have recently emerged as important players in innate immunity and antivirus. Here, we systematically analyzed mRNA expressions of representative TRIM molecules in human THP1-derived macrophages activated by different toll-like receptor (TLR) ligands. Twenty-nine TRIM members were highly induced (>3 fold) by one or more TLR ligands, among which 19 of them belong to TRIM C-IV subgroup. Besides TRIM21, TRIM22 and TRIM38 were shown to be upregulated by TLR3 and TLR4 ligands as previous reported, we identified a novel group of TRIM genes (TRIM14, 15, 31, 34, 43, 48, 49, 51 and 61) that were significantly up-regulated by TLR3 and TLR4 ligands. In contrast, the expression of TRIM59 was down-regulated by TLR3 and TLR4 ligands in both human and mouse macrophages. The alternations of the TRIM proteins were confirmed by Western blot. Finally, overexpression of TRIM59 significantly suppressed LPS-induced macrophage activation, whereas siRNA-mediated knockdown of TRIM59 enhanced LPS-induced macrophage activation. Taken together, the study provided an insight into the TLR ligands-induced expressions of TRIM family in macrophages.
Sujet(s)
Protéines adaptatrices de la transduction du signal/génétique , Macrophages péritonéaux/immunologie , Protéines membranaires/génétique , Récepteurs de type Toll/métabolisme , Protéines adaptatrices de la transduction du signal/métabolisme , Animaux , Lignée cellulaire , Cellules cultivées , Humains , Immunité innée , Lipopolysaccharides/pharmacologie , Activation des macrophages , Macrophages péritonéaux/effets des médicaments et des substances chimiques , Protéines membranaires/métabolisme , SourisRÉSUMÉ
With their unique optical properties and distinct Raman signatures, graphitic nanomaterials can serve as substrates for surface-enhanced Raman spectroscopy (SERS) or provide signal amplification for bioanalysis and detection. However, a relatively weak Raman signal has limited further biomedical applications. This has been addressed by encapsulating gold nanorods (AuNRs) in a thin graphitic shell to form gold graphitic nanocapsules. This step improves plasmon resonance, which enhances Raman intensity, and has the potential for integrating two-photon luminescence (TPL) imaging capability. However, changing the morphology of gold graphitic nanocapsules such that high quality and stability are achieved remains a challenge. To address this task, we herein report a confinement chemical vapor deposition (CVD) method to prepare the construction of AuNR-encapsulated graphitic nanocapsules with these properties. Specifically, through morphological modulation, we (1) achieved higher plasmon resonance with near-IR incident light, thus achieving greater Raman intensity, and (2) successfully integrated two-photon luminescence dual-modal (Raman/TPL) bioimaging capabilities. Cancer-cell-specific aptamers were further modified on the AuNR@G graphitic surface through simple, but strong, π-π interactions to achieve imaging selectivity through differential cancer cell recognition.
Sujet(s)
Or/composition chimique , Graphite/composition chimique , Imagerie multimodale/méthodes , Nanocapsules/composition chimique , Aptamères nucléotidiques/composition chimique , Survie cellulaire/effets des médicaments et des substances chimiques , Humains , Cellules MCF-7 , Microscopie confocale , Nanocapsules/toxicité , Nanotubes/composition chimique , Analyse spectrale Raman , Résonance plasmonique de surfaceRÉSUMÉ
Controlling and monitoring the drug delivery process is critical to its intended therapeutic function. Many nanocarrier systems for drug delivery have been successfully developed. However, biocompatibility, stability, and simultaneously tracing drugs and nanocarriers present significant limitations. Herein, we have fabricated a multifunctional nanocomposite by coating the gold nanorod (AuNR) with a biocompatible, superstable and fluorescent carbon layer, obtaining the AuNR@carbon core-shell nanocapsule. In this system, the carbon shell, originally obtained in aqueous glucose solutions and, therefore, biocompatible in physiological environments, could be simply loaded with cell-specific aptamers and therapeutic molecules through π-π interactions, a useful tool for cancer-targeted cellular imaging and therapy. Moreover, such a stable and intrinsic fluorescence effect of the AuNR@carbon enabled simultaneous tracking of released therapeutic molecules and nanocarriers under thermo-chemotherapy. The AuNR@carbons had high surface areas and stable shells, as well as unique optical and photothermal properties, making them promising nanostructures for biomedical applications.
Sujet(s)
Vecteurs de médicaments/composition chimique , Systèmes de délivrance de médicaments , Hyperthermie provoquée/méthodes , Nanocapsules/composition chimique , Antinéoplasiques/administration et posologie , Aptamères nucléotidiques , Carbone , Doxorubicine/administration et posologie , Or , Cellules HEK293 , Humains , Cellules MCF-7 , Nanoparticules métalliques/composition chimique , Nanotubes/composition chimiqueRÉSUMÉ
A group of chitin-binding proteins were isolated from tuberous roots of Raphanus sativus by affinity chromatography with deaminated regenerated chitin (Fig. 1). SDS-PAGE showed that there are at least five proteins in the sample (Fig. 2-b). Through carboxyl methyl-cellulose chromatography, two chitin-binding proteins with lysozyme activity, named as CBP1 and CBP2 (Fig. 3), were purified to homogeneity with the molecular weights of 26.9 kD and 24.8 kD respectively (Fig. 2-d, e). CBP1 and CBP2 were found to be bifunctional enzymes with activities of lysozyme and chitinase (Figs. 4, 5), but without chitosanase activity (Table 1). The CBP1 and CBP2 could be specifically absorbed by various forms of chitin, such as powdered, regenerated and colloidal forms chitin (Fig. 6). No disulfide bridge was observed in CBP1 and CBP2 by reduced/nonreduced one-dimensional SDS-PAGE (Fig. 7).