Toluene Dose-Response and Preliminary Study of Proteomics for Neuronal Cell Lines.

摘要

Toluene, a jet fuel component, is a potential contaminant in aircraft. Effects of toluene exposure include headache or fatigue. The objective of this study was to perform a preliminary investigation of the cellular and molecular effects of toluene on murine cell lines including Cath.a, C8-B4, CLU199, and HT22. The study tests two methods: direct dosing of toluene in the media and vapor exposure in glass chambers. To better define the complex signaling response to toluene, the two exposure models are characterized by cell viability assays and Tandem Mass Tags (TMT) liquid chromatography mass spectrometry (LC-MS/MS) quantitative proteomics proteome pathway analysis. Cell viability in response to direct dosing in a vehicle demonstrated a dose-dependent decrease in cell viability for all cell lines. Cytoscape 3.1.0 Reactome FI pathway analysis of Cath.a cells following direct dosing with 0.2 and 2 mM toluene revealed a dose-dependent toxicity increase of modulated protein nodes related to oxidative stress such as energy reserve metabolism, cell-death signaling, cytoskeletal rearrangement, DNA damage response, targeted protein degradation and apoptosis. While cell viability testing of all four cell lines exposed to 200 ppm toluene vapor for 1hour did not result in any detectable decrease of cell viability, proteomic analysis displayed minor signs of cellular stress with a majority of modulated proteins. These findings contribute to understanding the complex protein modifications within normally conserved proteins and toluene induced activation of cellular signaling pathways, which will aid in determining initiating mechanisms of brain toxicity from low level doses.