Voluven treatment could induce NLRP3 inflammasome-mediated pyroptosis of bone marrow-derived macrophages

Abstract

The aim of this study was to investigate the influence of Voluven on the NLRP3 inflammasome-mediated pyroptosis in bone marrow-derived macrophage (BMDM). Separated BMDMs were cultured and treated with different concentration of Voluven (0, 0.1, and 0.5 µg) dissolved in 10 µL 0.9% NaCl solution for 24 h. Both wild-type and nucleotide-binding domain and leucine-rich repeat protein 3 (NLRP3)–/– C57BL/6J mice ( n = 18) were intravenously injected with 0.2 mL of 0%, 5%, and 10% Voluven through femoral vein, respectively. Pyroptosis was inspected with flow cytometry. The mRNA levels of NLRP3 and caspase-1 were detected with quantitative real-time polymerase chain reaction (qRT-PCR). The levels of NLRP3, pro-caspase-1, and cleaved caspase-1 (p10) in serum were determined with Western blot. The expression of IL-17A in peripheral blood CD4+ T cells was analyzed with flow cytometry. The expression of cleaved caspase-1 (p10) in mice spleens was inspected with immunofluorescence. Compared with control group, the ratio of pyroptosis in all Voluven-treated groups rose significantly. The levels of NLRP3 and caspase-1 were increased after Voluven treatment. The expression of interleukin (IL)-17A in Voluven-treated CD4+ T cells was also increased, exhibiting a dose-dependent pattern. In wild-type mice, Voluven-treated mice had higher levels of IL-17A, NLRP3, and cleaved caspase-1 (p10) in a dose-dependent manner. The effects of Voluven were diminished in NLRP3–/– mice.