RESUMEN
BACKGROUND: The experimental autoimmune myocarditis (EAM) model is valuable for investigating myocarditis pathogenesis. M1-type macrophages and CD4+T cells exert key pathogenic effects on EAM initiation and progression. Baicalein (5,6,7-trihydroxyflavone, C15H10O5, BAI), which is derived from the Scutellaria baicalensis root, is a primary bioactive compound with potent anti-inflammatory and antioxidant properties. BAI exerts good therapeutic effects against various autoimmune diseases; however, its effect in EAM has not been thoroughly researched. PURPOSE: This study aimed to explore the possible inhibitory effect of BAI on M1 macrophage polarisation and CD4+T cell differentiation into Th1 cells via modulation of the JAK-STAT1/4 signalling pathway, which reduces the secretion of pro-inflammatory factors, namely, TNF-α and IFN-γ, and consequently inhibits TNF-α- and IFN-γ-triggered apoptosis in cardiomyocytes of the EAM model mice. STUDY DESIGN AND METHODS: Flow cytometry, immunofluorescence, real-time quantitative polymerase chain reaction (q-PCR), and western blotting were performed to determine whether BAI alleviated M1/Th1-secreted TNF-α- and IFN-γ-induced myocyte death in the EAM model mice through the inhibition of the JAK-STAT1/4 signalling pathway. RESULTS: These results indicate that BAI intervention in mice resulted in mild inflammatory infiltrates. BAI inhibited JAK-STAT1 signalling in macrophages both in vivo and in vitro, which attenuated macrophage polarisation to the M1 type and reduced TNF-α secretion. Additionally, BAI significantly inhibited the differentiation of CD4+T cells to Th1 cells and IFN-γ secretion both in vivo and in vitro by modulating the JAK-STAT1/4 signalling pathway. This ultimately led to decreased TNF-α and IFN-γ levels in cardiac tissues and reduced myocardial cell apoptosis. CONCLUSION: This study demonstrates that BAI alleviates M1/Th1-secreted TNF-α- and IFN-γ-induced cardiomyocyte death in EAM mice by inhibiting the JAK-STAT1/4 signalling pathway.
Asunto(s)
Apoptosis , Modelos Animales de Enfermedad , Flavanonas , Interferón gamma , Quinasas Janus , Miocarditis , Miocitos Cardíacos , Factor de Transcripción STAT1 , Transducción de Señal , Factor de Necrosis Tumoral alfa , Animales , Factor de Transcripción STAT1/metabolismo , Transducción de Señal/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Quinasas Janus/metabolismo , Ratones , Flavanonas/farmacología , Masculino , Interferón gamma/metabolismo , Apoptosis/efectos de los fármacos , Factor de Necrosis Tumoral alfa/metabolismo , Miocarditis/tratamiento farmacológico , Factor de Transcripción STAT4/metabolismo , Enfermedades Autoinmunes/tratamiento farmacológico , Ratones Endogámicos BALB C , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Scutellaria baicalensis/química , Células TH1/efectos de los fármacos , Diferenciación Celular/efectos de los fármacosRESUMEN
Although cotton dressing is one of the most commonly used wound management materials, it lacks antimicrobial and healing-promoting activity. This work developed a multilayer electroactive composite cotton dressing (Ag/Zn@Cotton/Paraffin) with exudate-activated electrical stimulation and antibacterial activity by the green and sustainable magnetron-sputtering and spraying methods. The inner hydrophilic layer of the cotton dressing was magnetron sputtered with silver/zinc galvanic couple arrays (Ag/Zn), which can be activated by wound exudate, generating an electrical stimulation (ES) into the wound. The Ag/Zn@Cotton showed efficient antibacterial activities against S. aureus and E. coli. Meanwhile, the paraffin-sprayed outer surface showed excellent antibacterial adhesion rates for S. aureus (99.82 %) and E. coli (97.92 %). The in vitro cell experiments showed that the ES generated by Ag/Zn@Cotton/Paraffin increased the migration of fibroblasts, and the in vivo mouse model indicated that the Ag/Zn@Cotton/Paraffin could enhance wound healing via re-epithelialization, inflammatory inhibition, collagen deposition, and angiogenesis. MTT method and live/dead staining showed that Ag/Zn@Cotton/Paraffin had no significant cytotoxic effects. This work may shed some light on designing and fabricating multi-functional electroactive composited dressings based on traditional biomedical textiles.
Asunto(s)
Fibra de Algodón , Staphylococcus aureus , Ratones , Animales , Escherichia coli , Parafina , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Cicatrización de Heridas , VendajesRESUMEN
Adjuvants are an indispensable component of vaccines, but there are few adjuvants for human vaccines. H2 receptor blockers, inhibiting gastric acid secretion, have immune enhancement effects. Ranitidine (RAN) is a water-soluble H2 receptor blocker, and whether it has an immune-enhancing effect is still unknown. In this study, flow cytometry, western blotting, and immunofluorescence methods were used to analyze whether RAN could activate macrophage polarization to the M1 phenotype in vivo and in vitro. Here, we found that the M1 inflammatory cytokine levels and surface markers in RAW264.7 cells were upregulated by NF-κB activation, possibly through the PI3K-Akt2 signaling pathway, after RAN treatment. Endocytic function was also enhanced by feedback regulation of Akt2/GSK3ß/Dynmin1 signaling. Furthermore, to evaluate the adjuvant function of RAN, we used OVA plus RAN as a vaccine to inhibit the growth of B16-OVA tumors in mice. We also found that in the RAN adjuvant group, macrophage polarization to M1, Th1 cell differentiation, and cytotoxic T lymphocyte (CTL) activation were significantly upregulated. The tumor growth of mice was inhibited, and the survival rate of mice was significantly improved. This study provides new evidence for the mechanism by which RAN activates the immune response and is expected to provide a new strategy for the research and development of tumor vaccine adjuvants.
Asunto(s)
Adyuvantes Inmunológicos , Macrófagos , Neoplasias , Ranitidina , Linfocitos T Citotóxicos , Animales , Humanos , Ratones , Adyuvantes Inmunológicos/farmacología , Adyuvantes Inmunológicos/uso terapéutico , Neoplasias/tratamiento farmacológico , Neoplasias/inmunología , FN-kappa B/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ranitidina/farmacología , Ranitidina/uso terapéutico , Células RAW 264.7 , Transducción de Señal , Linfocitos T Citotóxicos/efectos de los fármacos , Linfocitos T Citotóxicos/inmunología , Vacunas , Activación de Macrófagos/efectos de los fármacos , Activación de Macrófagos/inmunología , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Vacunas contra el Cáncer/inmunología , Vacunas contra el Cáncer/uso terapéuticoRESUMEN
Microglia/macrophage polarization modulation plays a key role in the pathogenesis of multiple sclerosis (MS)/experimental autoimmune encephalomyelitis (EAE). M1 microglia/macrophages secrete a variety of cytokines that cause inflammation and facilitate demyelination in the central nervous system (CNS). Baicalein (5,6,7-trihydroxyflavone, C15H10O5, BAI), a natural flavonoid isolated from the roots of the traditional Chinese medicine Scutellaria baicalensis Georgi, has been suggested to have a wide range of biological effects, including antioxidant, anti-inflammatory, and neuroprotective properties. In this study, flow cytometry, Western blotting, immunofluorescence and other methods were used to investigate whether BAI could reduce the demyelination and inflammatory response of the spinal cord in EAE mice induced by MOG35-55 and affect the polarization of spinal microglia/macrophages. Our results showed that BAI treatment delayed the onset of EAE and alleviated clinical symptoms, demyelination and inflammatory cell infiltration. Meanwhile, BAI inhibited the overactivation of M1 microglia/macrophages in vivo and in vitro, significantly decreased the expression of proinflammatory cytokines in M1 microglia/macrophages, and inhibited the activation of STAT1. Subsequently, molecular docking, pull-down and immunofluorescence experiments confirmed that BAI has the ability to bind to the SH2 domain of STAT1 and that BAI colocalizes with p-STAT1 in the cytoplasm rather than being transferred to the nucleus during inflammatory stimulation. This study showed that BAI might inhibit the polarization of microglia/macrophages to the M1 phenotype in EAE mice by targeting STAT1. This new discovery lays a theoretical and experimental foundation for the clinical application of BAI in the treatment of MS.