Role of caspase-8 and/or -9 as biomarkers that can distinguish the potential to cause toxic- and immune related-adverse event, for the progress of acetaminophen-induced liver injury

Acetaminophen (APAP) overdose can cause acute liver failure. Although it is well known that APAP-induced liver injury (AILI) is caused by toxic mechanism, recently it is also reported to be immune related. However, the detail of the mechanism has been unclear. Therefore, elucidation of the pathophysiology is required.In AILI model rats (800 mg/kg), the levels of AST, ALT and Caspase (C)-3/-8/-9 levels were measured. In in vitro study using human hepatocyte cells (FLC-4) and THP-1 cells, APAP (0.03-1.0 mM) were added to FLC-4 and the cell viability, C-9, cytochrome c, mitochondria membrane potential, and glutathione levels of FLC-4 and inflammasome activation of THP-1 were evaluated.In AILI model rats, the levels of AST and ALT were increased only at 12-24 h. C-3/-9 levels rose at 6-9 h, whereas C-8 level rose hours later, moreover, 24 h after; C-3/-8/-9 levels re-rose. In FLC-4 cells, cytochrome c was released from the mitochondria which is promoted by oxidative stress due to drug metabolism and C-9 was activated. Thus, AILI was caused mitochondrial damage by NAPQI as early reaction (first stage). In the next stage, inflammasomes of human antigen presenting cells, which released inflammatory cytokines were activated by damage-associated molecular patterns (DAMPs) released from damaged hepatocyte by APAP.It is confirmed that AILI includes immune related mechanism. Thereby, in case of N-acetylcysteine refractory, additional administration of steroid hormones should be effective and recommended as a novel strategy for AILI with immune related adverse event (irAE).