Combined cytotoxicity of co-exposure to aldehyde mixtures on human bronchial epithelial BEAS-2B cells

Abstract

Aldehydes are well-known air pollutants and often studied alone, while co-exposure to aldehyde mixtures is more common than single aldehydes. Unfortunately, it has been very little known about the (mechanism of) combined toxicity of aldehyde mixtures. Here, formaldehyde and acrolein were selected as the typical representatives of common aldehydes, and were used to explore to get in-depth insight into the mechanism of combined toxicity of aldehyde mixtures. The NOECs (non-observed effect concentrations) are 60 μmoL/L for formaldehyde, and 0.5 μmoL/L for acrolein, so acrolein is more toxic than formaldehyde. Formaldehyde and acrolein mixtures showed significant cytotoxicity and synergistic effects in a concentration/time-dependent way on BEAS-2B cells based on acute and chronic cytotoxicity assay. Acrolein was dominant in aldehyde mixtures in inducing cytotoxicity at environmentally relevant doses because of higher toxicity. Moreover, aldehyde mixtures significantly synergistically increased the intracellular reactive oxygen species (ROS), malondialdehyde (MDA) and lactate dehydrogenase (LDH) leakage, while caused an antagonistic effects on glutathione (GSH). Besides, formaldehyde could significantly potentiated the activation of environmental stress sensitive Nrf2 pathway induced by acrolein, even at doses at which formaldehyde treatment alone had no any response. Furthermore, as the downstream components of Nrf2 pathway, catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPX) and heme oxygenase-1 (HO-1) were significantly synergistically induced by formaldehyde and acrolein mixtures. Antioxidants N-acetylcysteine and reduced glutathione could significantly suppress the acute and chronic combined cytotoxicity of acrolein and formaldehyde mixtures, and changed their interactions (synergism) on cytotoxicity. Taken together, aldehyde mixtures have higher toxicity than that expected for additivity based on single aldehydes even at environmentally relevant concentrations, and the combined cytotoxicity may be enhanced through oxidative stress and the related Nrf2 pathway. Prolonged exposure to pollutants containing aldehyde mixtures through inhalation may have more serious threat to respiratory system in animal and human.