Project description:Purpose: Studies in mice have indicated that Tubuloside B has therapeutic potentials in various models of inflammatory diseases. We set out to analyze how Tubuloside B regulates transcription in LPS+IFN-γ-stimulated macrophages. Methods: RNA-seq was performed with two repetitions in RAW264.7 followed by treatment with Tubuloside B and LPS as well as IFN-γ. Conclusions: Tubuloside B inhibits the production of pro-inflammatory cytokines in activated macrophages.
Project description:This study aimed to investigate the mechanism of C2S-induced macrophagic inflammation and its’ effect on osteogenic differentiation of precursor cells. This study used the C2S (75-150 μg/ml) extract to induce macrophagic inflammation in RAW264.7 murine macrophages. RNA sequencing was preformed for analyzing mechanism of mitochondrial function and autophagy.
Project description:Macrophages are involved in the pathophysiology of many diseases as critical cells of the innate immune system. Pyroptosis is a form of macrophage death that induces cytokinesis of phagocytic substances in the macrophages, thereby defending against infection. Dimethyl itaconate (DI) is an analog of itaconic acid with anti-inflammatory effects. However, the effect of dimethyl itaconate on macrophage pyroptosis has not been elucidated clearly. Thus, the present study aimed to analyze the effect of DI treatment on a macrophage pyroptosis model (Lipopolysaccharide, LPS+Adenosine Triphosphate, ATP). The results showed that 0.25 mM DI ameliorated macrophage pyroptosis and downregulated interleukin (IL)-1β expression. Then, real-time quantitative polymerase chain reaction (RT-qPCR) was used to confirm the result of RNA-sequencing of the upregulated oxidative stress-related genes (Gclc and Gss) and downregulated inflammation-related genes (IL-12β and IL-1β). In addition, Gene Ontology (GO) enrichment analysis showed that differential genes were associated with transcript levels and DNA replication. Kyoto encyclopedia of genes and genomes (KEGG) enrichment showed that signaling pathways, such as tumor necrosis factor (TNF), Jak, Toll-like receptor and IL-17, were altered after DI treatment. N-acetyl-L-cysteine (NAC) reversed the DI effect on the LPS+ATP-induced macrophage pyroptosis and upregulated the IL-1β expression. Oxidative stress-related protein Nrf2 is involved in the DI regulation of macrophage pyroptosis. Taken together, these findings suggested that DI alleviates the pyroptosis of macrophages through oxidative stress.
Project description:The function of the mitochondrial protein CARD19 remains largely unknown. To begin to understand the molecular function of CARD19, we performed mass spectrometry-based proteomics to identify interacting partners of CARD19 in murine macrophages. We analyzed anti-MYC tag immunoprecipitates of 3xMYC-CARD19, expressed at approximately endogenous levels in raf-myc-immortalizd macrophages from a Card19-/- strain (Jackson Labs stock # 036463).