Fig. 10

ZnO NPs-induced inflammation is mediated by calcium-dependent pathways in BV2 cells. BV2 cells were preincubated for 1 h with A839977 (200 nM) and BAPTA-AM (20 μM) before ZnO NPs treatment. After the cells were treated with ZnO NPs at a dose of 30 μg/ml for 6 h. Summary of cell viability (a), TNF-α (b) and IL-1β (c) release and the expression of proinflammatory genes (f). The concentration of Ca²⁺ in BV2 cells, cells were stained with Ca²⁺ indicator Fluo4-AM (green) and DAPI (blue) (d). Scale bar represents 200 μm. Compare the mean fluorescence intensity of Ca²⁺ measured in BV2 cells (g). Total proteins of BV2 cells were extracted, and the levels of P₂X₇, NF-κB, ERK and p38 signaling pathway molecules were analyzed via Western Blot (e). The gray value was semiquantitative as shown in histogram (h). The 30 μg/mL ZnO NPs-induced cytotoxicity, increase in proinflammatory cytokine release and NF-κB, ERK and p38 phosphorylation was significantly inhibited by BAPTA-AM. Results shown as means ± SD from three independent experiments, compare with control group, *P < 0.05, **P < 0.01. Abbreviations: cont: control; ZnO: zinc oxide nanoparticles; P₂X₇: P₂X purinoceptor 7; DAPI: 4,6-diamino-2-phenyl indole SD: Standard deviation