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

AIM
To investigate the influence of voluven on the NLRP3 inflammasome mediated pyroptosis in bone marrow derived macrophage (BMDM).


METHODS
Separated BMDMs were cultured and treated with different concentration of voluven (0, 0.1, 0.5 µg) dissolving in 10 µL 0.9% Nacl solution for 24 h. Both wildtype and NLRP3-/- C57BL/6J mice (n=18) were intravenously injected with 0.2 mL 0, 5%, 10 % voluven through femoral vein, respectively. Pyroptosis were inspected with flow cytometry. The mRNA levels of NLRP3 and caspase-1 were detected with 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 were analyzed with flow cytometry. The expression of cleaved-caspase-1 (p10) in mice spleens were inspected with immunofluorescence.


RESULTS
Compared with control group, the ratio of pyroptosis in voluven treated groups were all significantly risen. The levels of NLRP3 and caspase-1 were increased after voluven treatment. The expression of IL-17A in voluven treated CD4+ T cells was also increased, exhibiting a dose-dependent pattern. In wildtype 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.