Project description:The p38MAPK downstream targets MK 2 and 3 are critical for the regulation of the macrophage response towards bacterial lipopolysaccharid (LPS). The extent these two kinases act cooperatively in regulating LPS-induced inflammatory cytokine expression and their distinct regulatory effects are still unclear. To address this point, whole transcriptome analyses had been performed using bone marrow derived macrophages (BMDM) generated from MK2-/- or MK2/3-/- animals and from wildtype littermates. The BMDM were treated with 100 ng/ml LPS for 360 minutes or with medium as control. Thereafter, total mRNA was isolated and subjected to array analysis using the SurePrint G3 Mouse 8x60K Microarray Kit from Agilent.
Project description:The p38MAPK downstream targets MK 2 and 3 are critical for the regulation of the macrophage response towards bacterial lipopolysaccharid (LPS). The extent these two kinases act cooperatively in regulating LPS-induced inflammatory cytokine expression and their distinct regulatory effects are still unclear. To address this point, whole transcriptome analyses had been performed using bone marrow derived macrophages (BMDM) generated from MK2-/- or MK2/3-/- animals and from wildtype littermates. The BMDM were treated with 100 ng/ml LPS for 120 minutes or with medium as control. Thereafter, total mRNA was isolated and subjected to array analysis using the SurePrint G3 Mouse 8x60K Microarray Kit from Agilent.
Project description:BACKGROUND Sepsis is an organ dysfunction characterized by systemic inflammatory response. Micro(mi)ribonucleic acids take part in the regulation of the inflammatory response in many conditions. However, the role and mechanism of miR-106a and anoctamin 1 (ANO1) in the inflammatory response in sepsis remain largely unknown. MATERIAL AND METHODS The serum samples were collected from 31 sepsis patients and healthy volunteers. Lipopolysaccharide (LPS)-treated RAW264.7 cells were used for the study in vitro. The inflammatory response was investigated via interleukin-6 and tumor necrosis factor-alpha levels using quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay. The expression abundances of miR-106a and ANO1 were detected via qRT-PCR or western blot. The target association between miR-106a and ANO1 was explored using dual-luciferase reporter analysis. RESULTS The inflammatory response was trigged in sepsis and LPS-treated RAW264.7 cells. miR-106a expression was enhanced and ANO1 declined in sepsis and LPS-treated RAW264.7 cells. Overexpression of ANO1 suppressed the inflammatory response and knockdown of ANO1 promoted the inflammatory response in RAW264.7 cells. ANO1 was directly targeted via miR-106a, and miR-106a reversed ANO1-mediated inflammatory inhibition in LPS-treated RAW264.7 cells. CONCLUSIONS MiR-106a regulated LPS-induced inflammatory response by targeting ANO1 in RAW264.7 cells, indicating the potential value of miR-106a for treatment of inflammatory diseases, including sepsis.
