Research Paper Volume 14, Issue 21 pp 8745—8762

Figure 4. Combination of NLRP3 inhibitor and Golph3 inhibitor ameliorates HG-caused microglia neuroinflammation. (A) The 3D structure of NLRP3 with zafirlukast; (B) The 3D structure of Golph3 with bromocriptine, the structure of zafirlukast/NLRP3 complex and Golph3/bromocriptine for molecular dynamic simulation were obtained by a standard docking procedure for a rigid protein and a flexible zafirlukast performed with Autodock 4.0, the image was analyzed by Pymol software. (C) The cell viability of BV2 treated with zafirlukast detected by CCK8 kit; (D) The cell viability of BV2 treated with bromocriptine detected by CCK8 kit; (E, F) qPCR detection of the IL-1β and IL-6 transcription level of BV2 cells treated with ZAF, BRO, and combination of ZAF and BRO under HG condition. (G, H) ELISA detection of IL-1β and IL-6 in BV2 cells treated with ZAF, BRO, and combination of ZAF and BRO under HG condition. (I) Western blot analysis of NLRP3, caspase1 and IL-1β in BV2 cells treated with ZAF, BRO, and combination of ZAF and BRO under HG condition. (J) Western blot analysis of Golph3, GM130, Vimentin in BV2 cells treated with ZAF, BRO, and combination of ZAF and BRO under HG condition. GAPDH was used as an internal control for normalization. Data represent means ± SEM of 3 independent experiments. * p ≤ 0.05, ** p ≤ 0.01, and *** p ≤ 0.001 according to two-way ANOVA with Bonferroni's post hoc test.