Isocyanates induces DNA damage, apoptosis, oxidative stress, and inflammation in cultured human lymphocytes

J Biochem Mol Toxicol. Nov-Dec 2008;22(6):429-40. doi: 10.1002/jbt.20260.

Abstract

Isocyanates, a group of low molecular weight aromatic and aliphatic compounds containing the isocyanate group (-NCO), are important raw materials with diverse industrial applications; however, pathophysiological implications resulting from occupational and accidental exposures of these compounds are hitherto unknown. Although preliminary evidence available in the literature suggests that isocyanates and their derivatives may have deleterious health effects including immunotoxicity, but molecular mechanisms underlying such an effect have never been addressed. The present study was carried out to assess the immunotoxic response of methyl isocyanate (MIC) on cultured human lymphocytes isolated from healthy human volunteers. Studies were conducted to evaluate both dose-dependent and time-course response (n = 3), using N-succinimidyl N-methylcarbamate, a surrogate chemical substitute to MIC. Evaluation of DNA damage by ataxia telangiectasia mutated (ATM) and gamma H2AX protein phosphorylation states; measure of apoptotic index through annexin-V/PI assay, apoptotic DNA ladder assay, and mitochondrial depolarization; induction of oxidative stress by CM-H2DCFDA and formation of 8-hydroxy-2' deoxy guanosine; levels of antioxidant defense system enzyme glutathione reductase; and multiplex cytometric bead array analysis to quantify the secreted levels of inflammatory cytokines, interleukin-8, interleukin-1beta, interleukin-6, interleukin-10, tumor necrosis factor, and interleukin-12p70 parameters were carried out. The results of the study showed a dose- and time-dependent response, providing evidence to hitherto unknown molecular mechanisms of immunotoxic consequences of isocyanate exposure at a genomic level. We anticipate these data along with other studies reported in the literature would help to design better approaches in risk assessment of occupational and accidental exposure to isocyanates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 8-Hydroxy-2'-Deoxyguanosine
  • Annexin A5 / metabolism
  • Antioxidants / metabolism
  • Apoptosis / drug effects*
  • Ataxia Telangiectasia Mutated Proteins
  • Cell Cycle Proteins / metabolism
  • Cells, Cultured
  • Cytokines / metabolism
  • DNA Damage*
  • DNA Fragmentation / drug effects
  • DNA-Binding Proteins / metabolism
  • Deoxyguanosine / analogs & derivatives
  • Deoxyguanosine / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Fluorescein-5-isothiocyanate
  • Glutathione Reductase / metabolism
  • Histones / metabolism
  • Humans
  • Inflammation / pathology*
  • Inflammation Mediators / metabolism
  • Isocyanates / pharmacology*
  • Lymphocytes / drug effects*
  • Lymphocytes / pathology*
  • Oxidative Stress / drug effects*
  • Phosphorylation / drug effects
  • Propidium
  • Protein-Serine-Threonine Kinases / metabolism
  • Reactive Oxygen Species / metabolism
  • Tumor Suppressor Proteins / metabolism

Substances

  • Annexin A5
  • Antioxidants
  • Cell Cycle Proteins
  • Cytokines
  • DNA-Binding Proteins
  • H2AX protein, human
  • Histones
  • Inflammation Mediators
  • Isocyanates
  • Reactive Oxygen Species
  • Tumor Suppressor Proteins
  • Propidium
  • 8-Hydroxy-2'-Deoxyguanosine
  • Glutathione Reductase
  • ATM protein, human
  • Ataxia Telangiectasia Mutated Proteins
  • Protein-Serine-Threonine Kinases
  • Deoxyguanosine
  • Fluorescein-5-isothiocyanate