ABSTRACT
OBJECTIVE@#To investigate the inflammatory effects of Cinobufotalin on monocytes in resting state and macrophages in activated state and its molecular mechanism.@*METHODS@#THP-1 cells were stimulated with Phorbol 12-myristate 13-acetate to induce differentiation into macrophages. Lipopolysaccharides was added to activate macrophages in order to establish macrophage activation model. Cinobufotalin was added to the inflammatory cell model for 24 h as a treatment. CCK-8 was used to detect cell proliferation, Annexin V /PI double staining flow cytometry was used to detect cell apoptosis, flow cytometry was used to detect macrophage activation, and cytometric bead array was used to detect cytokines. Transcriptome sequencing was used to explore the gene expression profile regulated by Cinobufotalin. Changes in the significantly regulated molecules were verified by real-time quantitative polymerase chain reaction and Western blot.@*RESULTS@#1∶25 concentration of Cinobufotalin significantly inhibited the proliferation of resting monocytes(P<0.01), and induced apoptosis(P<0.01), especially the activated macrophages(P<0.001, P<0.001). Cinobufotalin significantly inhibited the activation of macrophages, and significantly down-regulated the inflammatory cytokines(IL-6, TNF-α, IL-1β, IL-8) released by activated macrophages(P＜0.001). Its mechanism was achieved by inhibiting TLR4/MYD88/P-IκBa signaling pathway.@*CONCLUSION@#Cinobufotalin can inhibit the inflammatory factors produced by the over-activation of macrophages through TLR4/MYD88/P-IκBa pathway, which is expected to be applied to the treatment and research of diseases related to the over-release of inflammatory factors.
Subject(s)
Humans , Toll-Like Receptor 4/metabolism , Myeloid Differentiation Factor 88/genetics , Macrophages/metabolism , Cytokines/metabolism , Lipopolysaccharides/pharmacology , NF-kappa BABSTRACT
Myeloid-derived suppressor cells (MDSCs) are newly discovered cells derived from bone marrow that suppress immunity, playing an immunosuppressive role in tumors, infections, and autoimmune diseases. It has been confirmed in disease model studies that their immunosuppressive effect is mainly achieved by suppressing the function of T cells. At present, the treatment of infectious diseases is still a major obstacle, especially in the treatment of chronic infectious diseases. Studies have found that MDSCs have increased aggregation in infectious diseases. As MDSCs are gaining more attention in infectious diseases, their potential therapeutic effects may be further explored. This article mainly reviews the immunosuppressive mechanisms of MDSCs in infections such as bacteria, viruses, parasites and fungi and their relationship with the diseases.