Adenosine triphosphate-binding pocket inhibitor for mixed lineage kinase domain-like protein attenuated alcoholic liver disease via necroptosis-independent pathway

Figure 6 Mixed lineage kinase domain-like protein adenosine triphosphate-binding inhibitor attenuated steatosis and inflammation in alcoholic animal model. A: Overview of animal experiments. Mice were fed an ethyl alcohol (EtOH) diet for 10 days, and daily intraperitoneal injection of mixed lineage kinase domain-like protein (MLKL) adenosine triphosphate (ATP)-binding inhibitor (Compound-4) (10 mg/kg) was administered for 10 days. On the morning of 11^th day, EtOH was administered orally (5 g/kg), and euthanasia was performed nine hours later. Liver tissue and serum were collected; B: C57BL/6 mouse body weight through 10 days of experiment. The mice fed EtOH diet showed a trend of weight loss; n = 8; C: Liver tissue sections were stained with hematoxylin-eosin staining, 200 ×; D: Serum alanine transaminase, aspartate transaminase, and total bilirubin were measured (n = 8/group); E: Steatosis, inflammation, and fibrosis among EtOH and MLKL ATP-binding inhibitor (Compound-4) groups; F: CXCL1, CXCL2, ICAM, VCAM, MCP-1, and NLRP3 mRNA expression levels were measured via quantitative real-time polymerase chain reaction in liver tissue. P calculated compared to ethyl alcohol, mixed lineage kinase domain-like protein adenosine triphosphate-binding inhibitor (Compound-4). ^aP < 0.05. ^cP < 0.001. EtOH: Ethyl alcohol; HE: Hematoxylin-eosin staining; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; CPD4: Compound-4; ALT: Alanine transaminase; AST: Aspartate transaminase; ASH: Alcoholic steatohepatitis.