Toxicity mechanism of tetrabromobisphenol A to human respiratory system cells 16HBE and Beas2B

Abstract

Tetrabromobisphenol A (TBBPA) is a high-production chemical widely used in printed circuit boards and other polymeric materials for inflaming. As an environmental pollutant, it is widely found in various abiotic and biological media. Usually, the TBBPA could enter the body through skin contact, respiratory exposure and the dietary intake, among which respiratory exposure may be a most important exposure route. Previous studies reported that TBBPA possesses the endocrine, liver and kidney toxicity. However, few studies focused on the effects of TBBPA on respiratory toxicity. Therefore, it is urgent to study the toxic effects of TBBPA on respiratory system. In this study, human bronchial cells Beas2B and pulmonary epithelial cells 16HBE were used as subjects to study the exposure toxicity of TBBPA. Firstly, the viability and permeability of cells were assayed during the TBBPA exposure process. When TBBPA was in low concentration range (0−0.4 μmol/L), 24 h of exposure cannot significantly affect cell viability. When concentration of TBBPA was greater than 10 μmol/L, the inhibitory effect on cell activity was obviously observed. When concentration of TBBPA was up to 50 μmol/L, the cell viabilities of 16HBE and Beas2B cells decreased to 80.9% and 86.2%. It is worth noting that, as compared with bronchial epithelial cell 16HBE, lung epithelial cell Beas2B has a higher tolerance to TBBPA, which may be due to the different cell structures. In addition, the oxidation stress of cells was also determined by assaying the cell intracellular reactive oxygen species (ROSs), antioxidant enzymatic activity of superoxide dismutase (SOD) and catalase (CAT) during the whole expose duration. The results showed that the levels of intracellular ROSs obviously increased in both cells, indicating that peroxidation occurred in the cells during TBBPA exposure. Correspondingly, the enzymatic activities of SOD and CAT also increased, indicating that oxidative stress happened in cells. After that, flow cytometry was applied to analyze cell apoptosis. The results showed that partial apoptosis occurred after 24 h of TBBPA exposure, indicating that TBBPA could induce cell apoptosis. Besides, with the increase of exposure concentration, the apoptosis rate of the two kinds of cells also increased. Furthermore, the q-PCR was used to determine the apoptosis-related gene expression. The results revealed that the expression levels of apoptosis-related genes p53, Survivin , bax , bcl-2 , caspase-3 , caspase-9 were all up-regulated to different degrees, further confirming that cells were apoptotic caused by TBBPA. Further, compared with the 12-h exposure data, 24-h exposure would stimulate more intense expression up-regulation. In all, this research mainly investigated the toxicity of typical generation bromide flame retardants TBBPA on human bronchial epithelial cells 16 HBE and lung epithelial cells Beas2B. Results showed that the exposure of TBBPA could cause damage to cell viability and elicit oxidative stress in cells, which could further induce cell apoptosis. The results of cytotoxicity related to human respiratory system in this work can provide a scientific basis for revealing the health risk of toxic organic pollutants to human respiratory system.