New insights in acetaminophen toxicity: HMGB1 contributes by itself to amplify hepatocyte necrosis in vitro through the TLR4-TRIF-RIPK3 axis

Sci Rep. 2020 Mar 27;10(1):5557. doi: 10.1038/s41598-020-61270-1.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetaminophen / adverse effects*
  • Adaptor Proteins, Vesicular Transport / metabolism*
  • Cell Line
  • Cell Line, Tumor
  • Cells, Cultured
  • Chemical and Drug Induced Liver Injury / etiology
  • Chemical and Drug Induced Liver Injury / metabolism
  • HMGB1 Protein / metabolism*
  • Hepatocytes / drug effects
  • Hepatocytes / metabolism*
  • Humans
  • Liver / drug effects
  • Liver / metabolism
  • Necrosis / chemically induced
  • Necrosis / metabolism*
  • Receptor-Interacting Protein Serine-Threonine Kinases / metabolism*
  • Toll-Like Receptor 4 / metabolism*

Substances

  • Adaptor Proteins, Vesicular Transport
  • HMGB1 Protein
  • HMGB1 protein, human
  • TICAM1 protein, human
  • TLR4 protein, human
  • Toll-Like Receptor 4
  • Acetaminophen
  • RIPK3 protein, human
  • Receptor-Interacting Protein Serine-Threonine Kinases