Drug-induced liver injury from acetaminophen (APAP) overdose represents the leading cause of acute liver failure in developed nations. To date, the antioxidant N-acetyl cysteine is the sole first line treatment for APAP overdose, however, it is only effective at early stages of poisoning. In the liver, toxicity drives oxidative stress and mitochondrial dysfunction that leads to hepatocellular necrosis. Resident macrophages (Kupffer cells) respond to hepatic damage and initiate an inflammatory cascade of events including the recruitment of monocytes and neutrophils. Given the central importance of metabolic programs in regulating the hepatic immune environment/responses, we sought to uncover the role of AMP-activated protein kinase (AMPK) in APAP-induced liver injury. We first aimed to assess whether systemic activation of AMPK with MK-8722 could influence the damage and immune activation seen during APAP overdose. Secondly, we spatially interrogated the importance of AMPK in hepatocytes (deletion mediated by Alb-CreERT2) or myeloid cells (deletion driven by LysM-Cre). Finally, while myeloid AMPK deletion had minimal effect on hepatic necrosis or circulating markers of liver injury, we next aimed to specifically target resident macrophages with nanoparticles encapsulated with MK-8722. Interestingly, we observed increased infiltration of neutrophils in response to APAP in a myeloid AMPK-dependent manner. Dissecting the role of AMPK signaling in liver compartments will add to our understanding of how metabolic signaling underpins aspects of acute liver injury. Moreover, AMPK activation could serve as a therapeutic avenue for acute liver injury at later stages of overdose, either through systemic or targeted activation.