Human 3D multicellular microtissues: An upgraded model for the in vitro mechanistic investigation of inflammation-associated drug toxicity

Toxicol Lett. 2019 Sep 15;312:34-44. doi: 10.1016/j.toxlet.2019.05.004. Epub 2019 May 3.

Abstract

Inflammation is one of the factors that may increase the sensitivity of hepatic cells to acetaminophen (APAP) induced toxicity. To investigate the mechanisms, we exposed 3-dimensional (3D) Human Liver Microtissues, a co-culture of primary human hepatocytes (PHH) and Kupffer cells (KCs), to 0, 0.5 (low), 5 (median) and 10 mM (high dose) APAP for 24 h, with/without lipopolysaccharide (LPS). Microarray-technology was used to evaluate the transcriptome changes. In the presence of LPS, the median-dose of APAP is sufficient to inhibit the expression of respiratory chain- and antioxidant-related genes, suggesting the involvement of reactive oxygen species (ROS) and oxidative stress. Furthermore, the median- and high-dose of APAP inhibited the expression of Fc fragment receptor (FcγR)-coding genes, regardless of the presence of LPS. The toll-like receptor 4 (TLR4) expression, however, was continuously elevated after the LPS/APAP co-exposures, which may result in reduced KC-phagocytosis and unbalanced cytokine patterns. Compared to the treatment with LPS only, LPS/APAP co-exposures induced the production of interleukin (IL)-8, a pro-inflammatory cytokine, but suppressed the secretion of IL-6, a cytokine regulating hepatic regeneration, along with the increase in APAP dosages. In addition to the disrupted mitochondrial functions, the presence of LPS exacerbated APAP toxicity. These findings suggest that 3D Microtissues are a suitable model for the mechanistic exploration of inflammation-associated drug toxicity.

Keywords: 3D co-culture; Acetaminophen-inflammation interaction; Hepatotoxicity; Human Kupffer cells; Human primary hepatocytes; Transcriptomics.

MeSH terms

  • Acetaminophen / toxicity
  • Analgesics, Non-Narcotic / toxicity
  • Coculture Techniques
  • Cytokines / metabolism*
  • Gene Expression Regulation / drug effects
  • Hepatocytes / drug effects
  • Humans
  • Inflammation / metabolism*
  • Kupffer Cells / drug effects
  • Lipopolysaccharides / toxicity*
  • Receptors, IgG / genetics
  • Receptors, IgG / metabolism
  • Tissue Culture Techniques / methods*
  • Toll-Like Receptor 4 / genetics
  • Toll-Like Receptor 4 / metabolism
  • Transcriptome / drug effects

Substances

  • Analgesics, Non-Narcotic
  • Cytokines
  • Lipopolysaccharides
  • Receptors, IgG
  • TLR4 protein, human
  • Toll-Like Receptor 4
  • Acetaminophen