Figure 4.

P2Y2^−/− mice are protected from sepsis-induced systemic inflammation and liver injury. WT and P2Y2^−/− mice were subjected to CLP or sham surgery, and the liver and blood were harvested at 21 h post-CLP. A: representative images of H&E-stained liver sections, ×20 field of view. White arrow points to hemorrhagic necrosis and black arrows point to immune cell infiltrates. Black scale bar, 100 µm. B: serum ALT (n, WT-Sham, 4; WT-CLP, 12; P2Y2-Sham, 3; P2Y2-CLP, 5). C: serum bilirubin (n, WT-Sham, 4; WT-CLP, 12: P2Y2-Sham, 3; P2Y2-CLP, 6). D: representative liver sections stained for CD45⁺ leukocytes (×20 field of view). Black arrows point to leukocyte infiltration within hepatic parenchyma. Black scale bar, 100 µm. E: quantified by a stereological method at ×20 magnification by observers blinded to treatment groups. Bar diagram represents the analysis of five fields of view in each liver section (n, WT-Sham, 3, WT-CLP, 6; P2Y2-Sham, 3; P2Y2-CLP, 6). F: representative F4/80-stained liver sections (×20 field of view). Black arrows point to macrophage infiltration. Black scale bar, 100 µm. G–J: total protein extracts isolated from the livers harvested after CLP were analyzed by Western blotting with antibodies specific for iNOS (12 h post-CLP), NLRP3 (21 h post-CLP), GAPDH (protein loading control; n, WT-Sham, 3; WT-CLP, 4; P2Y2-Sham, 3; P2Y2-CLP, 4). Bar diagrams represent relative expression. K–P: multiplex assay of serum cytokines, TNFα, IL6, IL1β, IL10, MCP1, MKC (n, WT-Sham, 6; WT-CLP, 9–16; P2Y2-Sham, 6; P2Y2-CLP, 5–14). Means ±SD, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. n, number of biological replicates. CLP, cecal ligation and puncture; WT, wild type.