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Human bronchial epithelial cells exposed in vitro to diesel exhaust particles exhibit alterations in cell rheology and cytotoxicity associated with decrease in antioxidant defenses and imbalance in pro- and anti-apoptotic gene expression

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Abstract

Diesel exhaust particles (DEPs) from diesel engines produce adverse alterations in cells of the airways by activating intracellular signaling pathways and apoptotic gene overexpression, and also by influencing metabolism and cytoskeleton changes. This study used human bronchial epithelium cells (BEAS-2B) in culture and evaluates their exposure to DEPs (15ug/mL for 1 and 2 h) in order to determine changes to cell rheology (viscoelasticity) and gene expression of the enzymes involved in oxidative stress, apoptosis, and cytotoxicity. BEAS-2B cells exposed to DEPs were found to have a significant loss in stiffness, membrane stability, and mitochondrial activity. The genes involved in apoptosis [B cell lymphoma 2 (BCL-2 and caspase-3)] presented inversely proportional expressions (p = 0.05, p = 0.01, respectively), low expression of the genes involved in antioxidant responses [SOD1 (superoxide dismutase 1); SOD2 (superoxide dismutase 2), and GPx (glutathione peroxidase) (p = 0.01)], along with an increase in cytochrome P450, family 1, subfamily A, polypeptide 1 (CYP1A1) (p = 0.01). These results suggest that alterations in cell rheology and cytotoxicity could be associated with oxidative stress and imbalance between pro- and anti-apoptotic genes.

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Abbreviations

BEAS-2B:

Human bronchial epithelial cell line

BCL-2:

B cell lymphoma 2

DEP:

Diesel exhaust particles

PAHs:

Polycyclic aromatic hydrocarbons

LDH:

Lactate dehydrogenase

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

OMTC:

Optical magnetic twisting cytometry

*(f):

Complex apparent stiffness

Pa/nm:

Pascal per nanometer

η:

Hysteresivity

G*:

The complex shear modulus

g’:

The elastic modulus

g”:

The loss modulus

|G*|:

The complex modulus of G*

G:

|G*| at 0.75 Hz

SOD1:

Superoxide dismutase 1

SOD2:

Superoxide dismutase 1

GPx:

Glutathione peroxidase

CYP1A1:

Cytochrome P450, family 1, subfamily A, polypeptide 1

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Acknowledgments

This study was supported financially by the São Paulo Research Foundation (Fundação de Amparo à Pesquisa do Estado de São Paulo – FAPESP), Process No. 2011/50334-7. We would like to thank Cristina Grandal for editing the figures, and Dr. Carla Lima, and Marcio José Ferreira (Butantã Institute, São Paulo) and Thiago Franco Oliveira and Ana Paula Loureiro (University of São Paulo College of Pharmaceutical Sciences, Department of Clinical and Toxicological Analysis) for their help with cell cultures. Finally, we would like to thank Dr. Mário Luiz Ribeiro Monteiro (LIM33) and Roger Chammas (LIM24) for their critical analyses, suggestions, and technical support in the laboratory. Thais Mauad is funded by CNPq.

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Authors and Affiliations

  1. Laboratory of Experimental Air Pollution (LIM05), Department of Pathology, School of Medicine, University of São Paulo, Av. Dr. Arnaldo 455, 1°andar, sala 1150, Cerqueira César, São Paulo, SP, CEP:01246-903, Brazil

    Robson Seriani, Daniela Perroni Frias, Elnara Marcia Negri, Paulo Hilário Nascimento Saldiva, Thais Mauad & Mariangela Macchione

  2. Laboratory of Chronopharmacology, Department of Physiology, Institute of Bioscience, University of São Paulo, Rua do Matão - travessa 14, Cidade Universitária, 05508900, São Paulo, SP, Brazil

    Claudia Emanuele Carvalho de Souza

  3. Laboratory for Investigations in Ophthalmology (LIM-33), University of São Paulo Medical School São Paulo, Dr. Arnaldo 455, 5°andar, Cerqueira César, São Paulo, SP, CEP:01246-903, Brazil

    Paloma Gava Krempel & Monique Matsuda

  4. Heart Institute (InCor), Department of Cardiopulmonology, University of São Paulo Medical School, Av. Dr. Enéas de Carvalho Aguiar, 44, Cerqueira Cesar, 05403-000, Sao Paulo, SP, Brazil

    Aristides Tadeu Correia

  5. Laboratory of Microrheology and Molecular Physiology, Institute of Physics, University of São Paulo, Rua do Matão, Travessa R Número 187, Cidade Universitária, 05508-090, Sao Paulo, SP, Brazil

    Márcia Zotti Justo Ferreira & Adriano Mesquita Alencar

  6. FAM - Faculdades das Américas, Rua Augusta, 1508, 3°andar, São Paulo, SP, 01304-001, Brazil

    Robson Seriani

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  1. Robson Seriani
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Correspondence to Robson Seriani.

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Seriani, R., de Souza, C.E.C., Krempel, P.G. et al. Human bronchial epithelial cells exposed in vitro to diesel exhaust particles exhibit alterations in cell rheology and cytotoxicity associated with decrease in antioxidant defenses and imbalance in pro- and anti-apoptotic gene expression. Environ Sci Pollut Res 23, 9862–9870 (2016). https://doi.org/10.1007/s11356-016-6228-x

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