RESUMEN
BACKGROUND: C-reactive protein (CRP) is widely used as a sensitive biomarker for inflammation. Increasing evidence suggests that CRP plays a role in inflammation. High-mobility group box-1 (HMGB1), a primarily nuclear protein, is passively released into the extracellular milieu by necrotic or damaged cells and is actively secreted by monocytes/macrophages. Extracellular HMGB1 as a potent inflammatory mediator has stimulated immense curiosity in the field of inflammation research. However, the molecular dialogue implicated between CRP and HMGB1 in delayed inflammatory processes remains to be explored. METHODS AND RESULTS: The levels of HMGB1 in culture supernatants were determined by Western blot analysis and enzyme-linked immunosorbent assay in macrophage RAW264.7 cells. Purified CRP induced the release of HMGB1 in a dose- and time-dependent fashion. Immunofluorescence analysis revealed nuclear translocation of HMGB1 in response to CRP. The binding of CRP to the Fc gamma receptor in RAW264.7 cells was confirmed by fluorescence-activated cell sorter analysis. Pretreatment of cells with IgG-Fc fragment, but not IgG-Fab fragment, efficiently blocked this binding. CRP triggered the activation of p38MAPK and ERK1/2, but not Jun N-terminal kinase. Moreover, both p38MAPK inhibitor SB203580 and small interfering RNA significantly suppressed the release of HMGB1, but not the MEK1/2 inhibitor U-0126. CONCLUSION: We demonstrated for the first time that CRP, a prominent risk marker for inflammation including atherosclerosis, could induce the active release of HMGB1 by RAW264.7 cells through Fc gamma receptor/p38MAPK signaling pathways, thus implying that CRP plays a crucial role in the induction, amplification, and prolongation of inflammatory processes, including atherosclerotic lesions.
Asunto(s)
Proteína C-Reactiva/metabolismo , Proteína HMGB1/metabolismo , Inflamación/metabolismo , Macrófagos/metabolismo , Transducción de Señal/fisiología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Animales , Western Blotting , Línea Celular , Activación Enzimática/fisiología , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Ratones , Transporte de Proteínas/fisiología , ARN Interferente Pequeño , TransfecciónRESUMEN
High mobility group box-1 (HMGB1) is intranuclear architectural protein. Once HMGB1 released to extracellular, it has been implicated to act as a novel proinflammatory cytokine, called "Late mediator", in SIRS/sepsis. This danger signal transmits through receptor for advanced glycation end-products (RAGE). In this study, we hypothesized whether treatment of anti-HMGB1 or anti-RAGE antibody would block LPS-lethality in septic mice. We measured survival rate of septic mice with or without antibodies, resulting in significant improvement with antibodies-treated mice. LPS-induced acute lung injury was almost disappeared by antibody treatment, because these antibodies were inhibited HMGB1 and RAGE expression in lung. Treatment of anti-HMGB1 or anti-RAGE antibody seems to be valid therapy against SIRS/sepsis.
Asunto(s)
Anticuerpos/uso terapéutico , Dominios HMG-Box/inmunología , Receptores Inmunológicos/inmunología , Sepsis/terapia , Animales , Ratones , Receptor para Productos Finales de Glicación Avanzada , Síndrome de Respuesta Inflamatoria Sistémica/terapiaRESUMEN
A cyclic polyisoprenoid compound, geranylgeranylacetone (GGA), has been used as antiulcer drug. GGA is also a potent inducer of heat shock proteins (HSPs). HSPs are considered to induce an antiviral effect; however, the detailed mechanism is unknown. To determine whether GGA might show antiviral activity and what the mechanism is, the effect of GGA against influenza virus (strain PR8) infection in vivo and in vitro was investigated. The results demonstrated that GGA treatment strongly suppressed the deleterious consequences of PR8 replication and was accompanied by an increase in HSP70 gene expression in mice. Results from in vitro analyses demonstrated that GGA significantly inhibited the synthesis of PR8-associated proteins and prominently enhanced expression of human myxovirus resistance 1 (MxA) followed by increased HSP70 transcription. Moreover, GGA augmented the expression of an interferon-inducible double-strand RNA-activated protein kinase (PKR) gene and promoted PKR autophosphorylation and concomitantly alpha subunit of eukaryotic initiation factor 2 phosphorylation during PR8 infection. It is proposed that GGA-induced HSP70 has potent antiviral activity by enhancement of antiviral factors and can clinically achieve protection from influenza virus infection.