Gasdermin D-mediated hepatocyte pyroptosis expands inflammatory responses that aggravate acute liver failure by upregulating monocyte chemotactic protein 1/CC chemokine receptor-2 to recruit macrophages

Figure 2 Expression of pyroptosis pathway-associated proteins increases in a D-galactose/lipopolysaccharide-induced AML12 hepatocyte injury model. Intervention was performed using D-Galn (15 mmol/L) + LPS (100 µg/mL) for 0, 6, 12, and 24 h. A: Changes in the cell inhibition rate at 0, 6, 12, and 24 h evaluated by CCK8; B: Content of ALT in the supernatant; C: Expression of gasdermin D (GSDMD) protein detected under a confocal fluorescence microscope. Scale bars = 10 µm; D: Caspase 1/11, GSDMD-FL, and GSDMD-N protein expression evaluated by Western blot with GAPDH as the internal reference; E: Content of MCP1 in the cell culture supernatant. Data are presented as the mean ± square error. ^aP < 0.05, ^bP < 0.01, and ^eP < 0.001. ALT: Alanine aminotransferase; D-Galn/LPS: D-galactose/lipopolysaccharide; GSDMD-FL: Full-length GSDMD; GSDMD-N: Cleaved N-terminal fragment of GSDMD; MCP1: Monocyte chemotactic protein 1; DAPI: 4’,6-diamidino-2-phenylindole.