The NLRP3 Inflammasome and IL-1β Accelerate Immunologically Mediated Pathology in Experimental Viral Fulminant Hepatitis
Fig 5
NLRP3 inflammasome involves in regulating the pathogenesis of MHV-3-mediated hepatitis.
(A) Peritoneal exudative macrophages (PEMs) and RAW264.7 cells were infected with MHV-3 (MOI = 1) in vitro, and the expression of NLRP3 inflammasome complex components in the indicated time points was detected by western-blotting. (B) The transcription of NLRP3 and proCaspase-1 mRNA in liver tissues isolated from MHV-3 infected C57BL/6 WT mice was detected by qPCR (up penal), and the protein levels were measured by western-blotting (down penal). *p<0.05, NS: no significant difference. (C) The expression of IL-1β-p17, FGL2 and Bgp1 in NLRP3-/-, Caspase-1-/- and C57BL/6 WT livers at 0h and 72h post-infection was measured by western-blotting. (D) Liver fibrinogen deposition was analyzed by immunohistochemistry, the architecture was analyzed by H&E staining and cellular apoptosis was analyzed using TUNEL staining (left). Arrow indicates positive cells, blue color indicates nuclear staining with DAPI, scale bar 20 μm, n = 5 per group. Serum ALT and AST activities were determined with an AU5400 automatic biochemistry analyzer (right).*p<0.05, ** p<0.001, n = 5 per group, NS: no significant difference. (E) The virus titers in livers at 72h post-infection were analyzed by plaque assay (left), and their levels were compared by statistical analysis (right). *p<0.05, n = 5 per group. (F) The survival rate of virus infected mice was monitored for a total of 20 days. One representative of four experiments with similar results is shown. p<0.05 was considered significant different.