TY - JOUR
T1 - Differential proteomic analysis of mouse macrophages exposed to adsorbate-loaded heavy fuel oil derived combustion particles using an automated sample-preparation workflow Aerosols and Health
AU - Kanashova, Tamara
AU - Popp, Oliver
AU - Orasche, Jürgen
AU - Karg, Erwin
AU - Harndorf, Horst
AU - Stengel, Benjamin
AU - Sklorz, Martin
AU - Streibel, Thorsten
AU - Zimmermann, Ralf
AU - Dittmar, Gunnar
N1 - Funding Information:
This work was supported by funds of the Helmholtz virtual institute for complex Molecular Systems in Environmental Health (HICE). We thank Hanns Paur from the KIT, Karlsruhe, Germany for the loan of the CAROLA electrostatic precipitator and Patrick Beaudette as well as Daniel Perez-Hernandez for carefully reading the paper and fruitful discussions. We thank CTC and the Axel Semrau GmbH for the close collaboration in adapting the automation procedures for the CTC-PAL/Chronos setup.
Publisher Copyright:
© 2015 Springer-Verlag.
PY - 2015/8/27
Y1 - 2015/8/27
N2 - Ship diesel combustion particles are known to cause broad cytotoxic effects and thereby strongly impact human health. Particles from heavy fuel oil (HFO) operated ships are considered as particularly dangerous. However, little is known about the relevant components of the ship emission particles. In particular, it is interesting to know if the particle cores, consisting of soot and metal oxides, or the adsorbate layers, consisting of semi- and low-volatile organic compounds and salts, are more relevant. We therefore sought to relate the adsorbates and the core composition of HFO combustion particles to the early cellular responses, allowing for the development of measures that counteract their detrimental effects. Hence, the semi-volatile coating of HFO-operated ship diesel engine particles was removed by stepwise thermal stripping using different temperatures. RAW 264.7 macrophages were exposed to native and thermally stripped particles in submersed culture. Proteomic changes were monitored by two different quantitative mass spectrometry approaches, stable isotope labeling by amino acids in cell culture (SILAC) and dimethyl labeling. Our data revealed that cells reacted differently to native or stripped HFO combustion particles. Cells exposed to thermally stripped particles showed a very differential reaction with respect to the composition of the individual chemical load of the particle. The cellular reactions of the HFO particles included reaction to oxidative stress, reorganization of the cytoskeleton and changes in endocytosis. Cells exposed to the 280 °C treated particles showed an induction of RNA-related processes, a number of mitochondria-associated processes as well as DNA damage response, while the exposure to 580 °C treated HFO particles mainly induced the regulation of intracellular transport. In summary, our analysis based on a highly reproducible automated proteomic sample-preparation procedure shows a diverse cellular response, depending on the soot particle composition. In particular, it was shown that both the molecules of the adsorbate layer as well as particle cores induced strong but different effects in the exposed cells.
AB - Ship diesel combustion particles are known to cause broad cytotoxic effects and thereby strongly impact human health. Particles from heavy fuel oil (HFO) operated ships are considered as particularly dangerous. However, little is known about the relevant components of the ship emission particles. In particular, it is interesting to know if the particle cores, consisting of soot and metal oxides, or the adsorbate layers, consisting of semi- and low-volatile organic compounds and salts, are more relevant. We therefore sought to relate the adsorbates and the core composition of HFO combustion particles to the early cellular responses, allowing for the development of measures that counteract their detrimental effects. Hence, the semi-volatile coating of HFO-operated ship diesel engine particles was removed by stepwise thermal stripping using different temperatures. RAW 264.7 macrophages were exposed to native and thermally stripped particles in submersed culture. Proteomic changes were monitored by two different quantitative mass spectrometry approaches, stable isotope labeling by amino acids in cell culture (SILAC) and dimethyl labeling. Our data revealed that cells reacted differently to native or stripped HFO combustion particles. Cells exposed to thermally stripped particles showed a very differential reaction with respect to the composition of the individual chemical load of the particle. The cellular reactions of the HFO particles included reaction to oxidative stress, reorganization of the cytoskeleton and changes in endocytosis. Cells exposed to the 280 °C treated particles showed an induction of RNA-related processes, a number of mitochondria-associated processes as well as DNA damage response, while the exposure to 580 °C treated HFO particles mainly induced the regulation of intracellular transport. In summary, our analysis based on a highly reproducible automated proteomic sample-preparation procedure shows a diverse cellular response, depending on the soot particle composition. In particular, it was shown that both the molecules of the adsorbate layer as well as particle cores induced strong but different effects in the exposed cells.
KW - Dimethyl labeling
KW - Mitochondria
KW - Oxidative stress
KW - Polycyclic aromatic hydrocarbons
KW - SILAC
KW - Ship diesel exhaust particles
UR - http://www.scopus.com/inward/record.url?scp=84945478219&partnerID=8YFLogxK
U2 - 10.1007/s00216-015-8595-4
DO - 10.1007/s00216-015-8595-4
M3 - Article
C2 - 25772565
AN - SCOPUS:84945478219
SN - 1618-2642
VL - 407
SP - 5965
EP - 5976
JO - Analytical and Bioanalytical Chemistry
JF - Analytical and Bioanalytical Chemistry
IS - 20
ER -