Research Paper Volume 12, Issue 18 pp 18522—18544

GPA peptide inhibits NLRP3 inflammasome activation to ameliorate colitis through AMPK pathway

Figure 6. AMPK phosphorylation mediated the effects of GPA on decreasing ROS production. THP-1 cells were primed with LPS for 4 h, followed by GPA treatment 6 h before stimulation with ATP for 30 min, immunoblot analyzed of AMPK phosphorylation in cell lysate of THP-1 cells (A). Immunoblot analyzed of AMPK phosphorylation in colon tissues of mice (B). THP-1 cells were primed with LPS, followed by GPA, Compound C or GSK621 treatment 6 h before stimulation with ATP for 30 min. Levels of the ROS was measured in THP-1 cells (C). Cell death was measured by and LDH released (D). Immunoblot analyzed of IL-1β in supernatants and cell lysate of THP-1 cells (E). IL-1β in supernatants of THP-1 cells was detected by ELISA (F). Immunoblot analyzed of mitochondrial components of NLRP3 inflammasome in THP-1 cells (G). Data are presented as mean ± SD, three independent experiments. *p < 0.05, **p < 0.01 and ***p < 0.001.