Activation of the Nrf2 response by intrinsic hepatotoxic drugs correlates with suppression of NF-κB activation and sensitizes toward TNFα-induced cytotoxicity

Arch Toxicol. 2016 May;90(5):1163-79. doi: 10.1007/s00204-015-1536-3. Epub 2015 May 31.


Drug-induced liver injury (DILI) is an important problem both in the clinic and in the development of new safer medicines. Two pivotal adaptation and survival responses to adverse drug reactions are oxidative stress and cytokine signaling based on the activation of the transcription factors Nrf2 and NF-κB, respectively. Here, we systematically investigated Nrf2 and NF-κB signaling upon DILI-related drug exposure. Transcriptomics analyses of 90 DILI compounds in primary human hepatocytes revealed that a strong Nrf2 activation is associated with a suppression of endogenous NF-κB activity. These responses were translated into quantitative high-content live-cell imaging of induction of a selective Nrf2 target, GFP-tagged Srxn1, and the altered nuclear translocation dynamics of a subunit of NF-κB, GFP-tagged p65, upon TNFR signaling induced by TNFα using HepG2 cells. Strong activation of GFP-Srxn1 expression by DILI compounds typically correlated with suppression of NF-κB nuclear translocation, yet reversely, activation of NF-κB by TNFα did not affect the Nrf2 response. DILI compounds that provided strong Nrf2 activation, including diclofenac, carbamazepine and ketoconazole, sensitized toward TNFα-mediated cytotoxicity. This was related to an adaptive primary protective response of Nrf2, since loss of Nrf2 enhanced this cytotoxic synergy with TNFα, while KEAP1 downregulation was cytoprotective. These data indicate that both Nrf2 and NF-κB signaling may be pivotal in the regulation of DILI. We propose that the NF-κB-inhibiting effects that coincide with a strong Nrf2 stress response likely sensitize liver cells to pro-apoptotic signaling cascades induced by intrinsic cytotoxic pro-inflammatory cytokines.

Keywords: Drug-induced liver injury; Live-cell imaging; NF-κB signaling; Nrf2 activation; Oxidative stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Active Transport, Cell Nucleus
  • Chemical and Drug Induced Liver Injury / genetics
  • Chemical and Drug Induced Liver Injury / metabolism*
  • Chemical and Drug Induced Liver Injury / pathology
  • Computational Biology
  • Databases, Genetic
  • Dose-Response Relationship, Drug
  • Gene Expression Profiling / methods
  • Gene Expression Regulation / drug effects
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / genetics
  • Hep G2 Cells
  • Hepatocytes / drug effects*
  • Hepatocytes / metabolism
  • Hepatocytes / pathology
  • Humans
  • Liver / drug effects*
  • Liver / metabolism
  • Liver / pathology
  • NF-E2-Related Factor 2 / genetics
  • NF-E2-Related Factor 2 / metabolism*
  • Oxidoreductases Acting on Sulfur Group Donors / biosynthesis
  • Oxidoreductases Acting on Sulfur Group Donors / genetics
  • RNA Interference
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Recombinant Fusion Proteins / biosynthesis
  • Signal Transduction / drug effects
  • Time Factors
  • Transcription Factor RelA / genetics
  • Transcription Factor RelA / metabolism*
  • Transfection
  • Tumor Necrosis Factor-alpha / toxicity*


  • NF-E2-Related Factor 2
  • NFE2L2 protein, human
  • RELA protein, human
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Transcription Factor RelA
  • Tumor Necrosis Factor-alpha
  • Green Fluorescent Proteins
  • Oxidoreductases Acting on Sulfur Group Donors
  • SRXN1 protein, human