Methylisothiazolinone may induce cell death and inflammatory response through DNA damage in human liver epithelium cells

Environ Toxicol. 2018 Feb;33(2):156-166. doi: 10.1002/tox.22503. Epub 2017 Nov 7.

Abstract

Methylisothiazolinone (MIT) is a powerful biocide and preservative, which is widely used alone or in a 1:3 ratio with methylchloroisothiazolinone (MCIT) under the trade name of Kathons in the manufacture of numerous personal and household products. Considering that Kathons injected intravenously is distributed in the blood and then in the liver, we explored the toxic mechanism of MIT on human liver epithelium cells. At 24 h after exposure, MIT bound to the plasma membrane and the inner wall of vacuoles in the cells, and rupture of the cell membrane and nuclear envelop, autophagosome-like vacuoles formation and mitochondrial damage were observed. Cell viability dose-dependently decreased accompanying an increase of apoptotic cells, and the level of LDH, NO, IFN-gamma, IL-10 and IL-8, but not IL-1β, significantly increased in the culture media of cells exposed to MIT. Additionally, expression of autophagy-, membrane damage- and apoptosis-related proteins was notably enhanced, and the produced ATP level dose-dependently decreased with the reduced mitochondrial activity. Furthermore, the increased DNA damage and the decreased transcription activity were observed in MIT-treated cells. Meanwhile, the intracellular ROS level did not show dose-dependent change at the same time-point. Then we explored the role of autophagy in MIT-induced cytotoxicity by inhibiting or inducing the autophagic signal. Intriguingly, no additional cell death induced by autophagic modulation occurred when MIT was treated. Taken together, we suggest that MIT may induce multiple pathways of cell death and inflammatory response through DNA damage caused by rupture of the nuclear envelope.

Keywords: DNA damage; cell death; inflammation; methylisothiazolinone; nuclear envelope.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Apoptosis / drug effects*
  • Cell Line
  • DNA Damage / drug effects*
  • Enzyme-Linked Immunosorbent Assay
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Humans
  • Inflammation Mediators / metabolism*
  • Interferon-gamma / analysis
  • Interleukin-10 / analysis
  • Interleukin-8 / analysis
  • Liver / cytology
  • Microscopy, Electron, Transmission
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • Nitric Oxide / metabolism
  • Reactive Oxygen Species / metabolism
  • Thiazoles / toxicity*

Substances

  • Inflammation Mediators
  • Interleukin-8
  • Reactive Oxygen Species
  • Thiazoles
  • Interleukin-10
  • 2-methyl-4-isothiazolin-3-one
  • Nitric Oxide
  • Interferon-gamma
  • Adenosine Triphosphate