The cross-talk of NLRP3 inflammasome activation and necroptotic hepatocyte death in acetaminophen-induced mice acute liver injury

Abstract

Overdose acetaminophen (APAP) can result in severe liver injury, which is responsible for nearly half of drug-induced liver injury in western countries. Previous studies have found that there existed massive hepatocellular necrosis and severe inflammatory response in APAP-induced liver injury. However, the mechanistic linkage between necroptosis and NLRP3 inflammasome pathway in APAP-induced hepatotoxicity remains poorly understood. In order to investigate the relationship between inflammation and hepatocytes death in APAP hepatotoxicity, a time-course model for APAP hepatotoxicity in C57/BL6 mice was established by intraperitoneal (i.p) injection of 300 mg/kg APAP in this study. The activity of serum enzymes and pathological changes of APAP-treated mice were evaluated, and the critical molecules in necroptosis and NF-κB-NLRP3 inflammasome signaling pathway were determined by immunoblot and immunofluorescence analysis. The results demonstrated that APAP overdose resulted in a severe liver injury. Furthermore, the expression of critical molecules in NLRP3 inflammasome and necroptosis pathways peaked at 12–24 h, and then was decreased gradually, which is consistent with the pattern of pathological injury induced by APAP. Our further investigation found that the level of IL-1β in mouse liver was closely correlated with the level of phosphorylated MLKL following exposure to APAP. Furthermore, inhibition of necroptosis with necrostatin-1 significantly suppressed the activation of NLRP3 inflammasome signaling. Taken together, our results highlighted that the cross-talk between necroptosis and NLRP3 inflammasome played a critical role for promoting APAP-induced liver injury. Inhibition of the interaction of inflammation and necroptosis by pharmaceutical methods may represent a promising therapeutic strategy for APAP-induced liver injury.

Diacerein counteracts acetaminophen-induced hepatotoxicity in mice via targeting NLRP3/caspase-1/IL-1β and IL-4/MCP-1 signaling pathways

“…Necrotic hepatocytes release damage-associated molecular patterns (DAMPs) that activate the NLR family pyrin domain containing 3 (NLRP3) inflammasome, which cleaves procaspase-1, forming active caspase-1 (Roh and Sohn 2018 ; Li et al 2021 ). Caspase-1 then cleaves pro-interleukin-1 beta (pro-IL-1β), forming active IL-1β, which triggers the recruitment of neutrophils and monocytes to the liver (Woolbright and Jaeschke 2017 ; Li et al 2021 ; Seok et al 2021 ).…”

Roles of necroptosis in alcoholic liver disease and hepatic pathogenesis

“…Recent research reported that the expression of critical molecules in NLRP3 inflammasome was significantly higher in APAP‐treated mice than in control mice, and this difference disappeared following inhibition of necroptosis with Nec‐1. In addition, further investigation indicated that the level of IL‐1β in mouse liver was closely consistent with the level of p‐MLKL in response to APAP exposure, as evidenced by co‐localization of p‐MLKL and IL‐1β 74 . Tao's study demonstrated that RIPK1 activity was promoted in hepatic macrophages after HFD feeding, but hepatic inflammation was obviously alleviated in RIPK1 kinase‐dead (RIPK1 K45A/K45A ) mice compared with WT mice, implying that RIPK1 is required for controlling hepatic macrophage infiltration and activation 75 .…”

“…In addition, further investigation indicated that the level of IL‐1β in mouse liver was closely consistent with the level of p‐MLKL in response to APAP exposure, as evidenced by co‐localization of p‐MLKL and IL‐1β. 74 Tao's study demonstrated that RIPK1 activity was promoted in hepatic macrophages after HFD feeding, but hepatic inflammation was obviously alleviated in RIPK1 kinase‐dead (RIPK1 K45A/K45A ) mice compared with WT mice, implying that RIPK1 is required for controlling hepatic macrophage infiltration and activation. 75 Also, the results obtained in both bone marrow–derived macrophages (BMDMs) and primary Kupffer cells (KCs) further confirmed the hypothesis that RIPK1 plays a crucial role in mediating inflammasome activity.…”

Pathophysiology and Treatment Options for Hepatic Fibrosis: Can It Be Completely Cured?

“…In the liver, activated KCs stimulate NLRP3, which results in a wide range of immune responses including production of pro-inflammatory cytokines and chemokines that subsequently induce cell death [ 127 ]. NLRP3 initiates inflammation, leading to tissue damage and fibrosis in various conditions including alcoholic steatohepatitis, DILI, and NASH, through IL-17 and TNF-α [ 128 ]. NLRP3 mutant mice have been shown to have higher expression of these cytokines that were mainly produced by the infiltrating cells.…”