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 3 Gasdermin D knockdown by shRNA decreases the expression of monocyte chemotactic protein 1 and CC chemokine receptor-2 in a D-galactose/lipopolysaccharide-induced AML12 hepatocyte injury model. Control shRNA and gasdermin D (GSDMD) shRNA were transfected into AML12 using Lipofectamine 3000, and the intervention was performed using D-galactose (15 mmol/L)/lipopolysaccharide (100 µg/L) for 24 h. A: Expression changes of GSDMD protein evaluated by Western blot; B: Protein expression levels of monocyte chemotactic protein 1 (MCP1) and CC chemokine receptor-2 evaluated by Western blot; C: Cell inhibition rate evaluated by CCK8 assay; D: Contents of alanine aminotransferase and MCP1 in the cell supernatant. Data are presented as the mean ± square error. ^aP < 0.05, ^bP < 0.01, and ^eP < 0.001. D-Galn/LPS: D-galactose/lipopolysaccharide; GSDMD: Gasdermin D; ALT: Alanine aminotransferase; MCP1: Monocyte chemotactic protein 1; CCR2: CC chemokine receptor-2.