New insights in acetaminophen toxicity: HMGB1 contributes by itself to amplify hepatocyte necrosis in vitro through the TLR4-TRIF-RIPK3 axis
New insights in acetaminophen toxicity: HMGB1 contributes by itself to amplify hepatocyte necrosis in vitro through the TLR4-TRIF-RIPK3 axis
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DOI：
10.1038/s41598-020-61270-1
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发表时间：
2020-03-27
影响因子：
4.6
通讯作者：
Gustot, Thierry
中科院分区：
文献类型：
--
作者：
Minsart, Charlotte;Liefferinckx, Claire;Gustot, Thierry
HMGB1的胞外释放参与了扑热息痛诱导的肝损伤。在这一毒性过程中，HMGB1作为一种危险相关的分子模式发挥作用，但其作用机制和靶细胞尚未完全确定。在此，我们在体外研究了HMGB1在放大扑热息痛诱导的肝细胞坏死过程中的作用。使用培养的HepaRG细胞、原代人肝细胞和选择性化学抑制剂，我们评估了扑热息痛的毒性。我们证实了醋氨酚的加入诱导了HepaRG细胞死亡和HMGB1的释放。我们发现，抑制HMGB1可以减少扑热息痛诱导的HepaRG细胞死亡，这表明HMGB1具有前馈效应。我们提供了第一个证据，表明重组人HMGB1(RhHMGB1)暴露于HepaRG细胞也会导致细胞死亡。此外，我们发现对乙酰氨基酚和重组人HMGB1均通过依赖RIPK3的机制诱导程序性HepaRG细胞坏死。通过使用TLR4封闭抗体，我们证实了对乙酰氨基酚和rhHMGB1诱导的HepaRG细胞死亡的减少。此外，抑制TRIF，已知可以诱导依赖RIPK3的细胞死亡，减少了rhHMGB1诱导的HepaRG细胞死亡。我们的数据支持扑热息痛应激肝细胞释放HMGB1通过TLR4-TRIF-RIPK3途径引起邻近肝细胞的坏死。这项体外研究为HMGB1在对乙酰氨基酚诱导的毒性放大中的作用提供了新的见解。
Extracellular release of HMGB1 contributes to acetaminophen-induced liver injury. HMGB1 acts as a danger-associated molecular patterns during this toxic process but the mechanisms of action and targeted cells are incompletely defined. Here we studied, in vitro, the role of HMGB1 in amplifying the acetaminophen-induced hepatocyte necrosis process. Using cultured HepaRG cells, primary human hepatocytes and selective chemical inhibitors we evaluated acetaminophen-induced toxicity. We confirmed that addition of acetaminophen induced HepaRG cell death and HMGB1 release. We showed that inhibition of HMGB1 decreased acetaminophen-induced HepaRG cell death, suggesting a feedforward effect. We provide the first evidence that exposure of HepaRG cells to recombinant human HMGB1 (rhHMGB1) also resulted in cell death. Moreover, we found that both acetaminophen and rhHMGB1 induced programmed HepaRG cell necrosis through a RIPK3-dependent mechanism. By using TLR4 blocking antibody, we demonstrated the reduction of the HepaRG cell death induced by acetaminophen and rhHMGB1. Furthermore, inhibition of TRIF, known to induce a RIPK3-dependent cell death, reduced rhHMGB1-induced HepaRG cell death. Our data support that released HMGB1 from acetaminophen-stressed hepatocytes induced necrosis of neighboring hepatocytes by TLR4-TRIF-RIPK3- pathway. This in vitro study gives new insights in the role of HMGB1 in the amplification of acetaminophen-induced toxicity.