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.

过量服用对乙酰氨基酚（APAP）会导致严重的肝损伤，在西方国家，近一半的药物性肝损伤是由这种物质引起的。以往的研究发现APAP引起的肝损伤存在大量肝细胞坏死和严重的炎症反应。然而，在 APAP 诱导的肝毒性中，坏死性凋亡和 NLRP3 炎性体通路之间的机制联系仍然知之甚少。为了研究炎症与肝细胞死亡在 APAP 肝毒性中的关系，本研究通过腹腔 (ip) 注射 300 mg/kg APAP 建立了 C57/BL6 小鼠 APAP 肝毒性的时程模型。评估APAP处理小鼠的血清酶活性和病理变化，并通过免疫印迹和免疫荧光分析确定坏死性凋亡和NF-κB-NLRP3炎性体信号通路的关键分子。结果表明，APAP 过量导致严重的肝损伤。此外，NLRP3炎性体和坏死性凋亡通路中关键分子的表达在12-24小时达到峰值，然后逐渐下降，这与APAP诱导的病理损伤模式一致。我们的进一步调查发现，小鼠肝脏中 IL-1β 的水平与暴露于 APAP 后磷酸化 MLKL 的水平密切相关。此外，用 necrostatin-1 抑制 necroptosis 显着抑制了 NLRP3 炎性体信号传导的激活。综合起来，我们的结果强调，坏死性凋亡和 NLRP3 炎症小体之间的串扰在促进 APAP 诱导的肝损伤中发挥了关键作用。通过药物方法抑制炎症和坏死性凋亡的相互作用可能代表了 APAP 诱导的肝损伤的一种有前途的治疗策略。